2021 한국생물공학회 춘계학술발표대회

및 국제심포지엄

2021 KSBB Spring Meeting and

International Symposium

2021. 4. 14(수) ~ 16(금)

April 14(Wed) ~ 16(Fri), 2021

2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

Plenary Lecture

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S111 Ideonella sakaiensis, PETase and MHETase: From Identification of Microbial PET Degradation to
the Current Situation

Kohei ODA
Kyoto Institute of Technology, Matsugasaki, Kyoto, Japan
Corresponding Author Email : [email protected]

Poly(ethylene terephthalate) (PET) is used extensively worldwide, and its accumulation in the environment
threatens ecosystems. We succeeded in isolation from a microbial consortium of Ideonella sakaiensis 201-F6,
microorganism that is able to use PET as its energy and carbon sources, and characterization of the key
enzymes, PETase and MHETase (Yoshida et al., Science 2016). This discovery accelerated research on not
only these enzymes, but also on cutinases known to degrade PET. As a result, cutinase LCC is close to its use
for industrial-scale treating of waste PET. In this symposium, the current status of this research area will be
introduced.

Keywords : Poly(ethylene terephthalate) (PET), Microbial consortium, Ideonella sakaiensis, PET hydrolase
(PETase), Mono(2-hydroxyethyl) terephthalic acid hydrolase (MHETase), PET hydrolytic enzyme (PHE)
References
1. Shosuke Yoshida, Kazumi Hiraga, Toshihiko Takehana, Ikuo Taniguchi, Hironao Yamaji, Yasuhito Maeda,
Kiyotsuna Toyohara, Kenji Miyamoto, Yoshiharu Kimura, Kohei Oda. A bacterium that degrades and
assimilates poly(ethylene terephthalate), Science, 351, 1196-1199 (2016).
2. Shosuke Yoshida, Kazumi Hiraga, Toshihiko Takehana, Ikuo Taniguchi, Hironao Yamaji, Yasuhito Maeda,
Kiyotsuna Toyohara, Kenji Miyamoto, Yoshiharu Kimura, Kohei Oda. Response to comment on “A bacte-
rium that degrades and assimilates poly(ethylene terephthalate)”, Science, 353, 759-e (2016).
3. Ikuo Taniguchi, Shosuke Yoshida, Kazumi Hiraga, Kenji Miyamoto, Yoshiharu Kimura, Kohei Oda. Bio-
degradation of PET: Current status and application aspects, ACS Catalysis, 9, 4089-4105 (2019).
4. Kazumi Hiraga, Ikuo Taniguchi, Shosuke Yoshida, Yoshiharu Kimura, Kohei Oda. Biodegradation of waste
PET A sustainable solution for dealing with plastic pollution, EMBO Reports, e49365 (2019).
5. Shosuke Yoshida, Kazumi Hiraga, Ikuo Taniguchi, Kohei Oda. Methods in Enzymology, Vol. 648,
Enzymatic Plastic Degradation, Chapter 9. Ideonella sakaiensis, PETase, and MHETase: From identi-
fication of microbial PET degradation to enzyme characterization, edited by Gert Weber, Uwe T.
Bornscheur and Ren Wei, Academic Press (2021).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S112 Engineering Enzyme Therapeutics for Cancer Immunotherapy and Inborn Errors of Metabolism

George GEORGIOU
Depts of Chemical Engineering, Molecular Biosciences, Biomedical Engineering, and Oncology, University of
Texas, Austin, USA
Corresponding Author Email : [email protected]

Cancer is predicated on the suppression of adaptive immune responses. A key mechanism for immune escape in
cancer arises from genetic changes and/or metabolic re-programming that result in the aberrant accumulation of
metabolites that potently suppress lymphocyte function. Our lab has pioneered the concept of immune check-
point enzyme therapy whereby systemic administration of pharmacologically optimized, human engineered
enzymes is employed to activate the immune system leading to cancer eradication. We have invented and led
the development of 5 such engineered enzymes that are now either in clinical or late-stage preclinical
evaluation. In this presentation I will describe the mechanism of action for two of these checkpoint enzymes and
the respective protein engineering campaigns that led to the design of the clinical candidate molecules for
human studies. I will also briefly mention our related work on engineered human enzymes for the treatment of
inborn error of metabolism diseases.

References
1. Cramer, S.L., et al., “Systemic Depletion of L-Cyst(e)ine with Cyst(e)inase Increases Reactive Oxygen
Species and Suppresses Tumor Growth,” Nature Medicine 23120-127 (2017).
2. Triplett, T.A. et al., “Reversal of Indoleamie 2,3-Dioxygenase-Mediated Cancer Immune Suppression by
Systemic Kynurenine Depletion with a Therapeutic Enzyme,” Nature Biotechnology 36:758-764 (2018).
3. Wang, W., et al., “CD8+ T Cells Regulate Tumour Ferroptosis During Cancer Immunotherapy” Nature
569:270-274 (2019).
4. Lu, W-C. et al., “Enzyme-Mediated Depletion of Serum L-Met Abrogates Prostate Cancer Growth via
Multiple Mechanisms without Evidence of Systemic Toxicity, “Proc. Natl. Acad. Sci. USA 117:532-540
(2020).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S113 Emergence of Genome Engineering in the 2020s

Bernhard PALSSON
University of California, San Diego, 9500 Gilman Drive, La Jolla, CA, USA
Corresponding Author Email : [email protected]

Background: We have reached the point in time where we can realistically contemplate the design of entire
genomes. This will open a new and fundamental chapter in the history of engineering applied to life science.
One can thus hypothesize that a new fundamental engineering discipline will emerge during the 2020s, and most
likely the first Departments of Genome Engineering will be in operation at leading universities around the world
by 2030.

Foundations of engineering disciplines: Engineering disciplines are built on similar foundations, including,
definition, measurement, modeling (mechanistic and phenomenological), construction, and control. We now
have genome-wide capabilities representing these foundations for Genome Engineering.

Need for modeling and prediction: The need to integrate knowledge types into big data analytics, generally
referred to as explanatory-artificial-intelligence, is growing. Existing genome-scale modeling tools are well
positioned to play an important role, but they must be integrated with big data analytics. This talk will describe
progress with three approaches to such knowledge enrichment: 1) the use of Independent Component Analysis
(ICA) to define independently modulated sets of genes in bacterial transcriptomes, 2) the use of pangenome
analysis for the thousands of bacterial genome sequences being generated, and 3) the use of machine learning
methods for the analysis of binary phenotypes, such as antimicrobial resistance.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S114 Type III CRISPR-Cas - from Structure-function Analyses to Diagnostics

Jurre A. STEENS, Yifan ZHU, John VAN DER OOST, Raymond H.J. STAALS
Laboratory of Microbiology, Wageningen University and Research, 6708 WE, Wageningen, The Netherlands
Corresponding Author Email : [email protected]

Characteristic properties of type III CRISPR-Cas systems include recognition of target RNA (rather than DNA)
and the subsequent induction of a multifaceted immune response. This involves sequence-specific cleavage of a
target RNA and production of cyclic oligoadenylate (cOA) second messenger molecules that may trigger
dormancy or cell death. In this study, we discovered that a largely exposed seed region at the 3’ end of the
crRNA is essential for target RNA binding and cleavage, whereas base pairing at a unique region at the 5’ end
of the guide is required to trigger cOA production. Moreover, we uncovered that the natural variation in the
composition of type III complexes within a single host results in different guide lengths, and hence variable seed
regions. This shifting seed may prevent escape by invading genetic elements, while controlling cOA production
very tightly to prevent unnecessary damage to the host. Lastly, we used these findings to develop a new
diagnostic tool, named SCOPE, which was used for the specific detection of SARS-CoV-2 from human nasal
swab samples, showing sensitivities in the atto-molar range.

Keywords : CRISPR-Cas, type III, Cmr complex, Cas10, seed, cyclic oligoadenylates, diagnostics, SARS-
CoV-2
References
1. Steens, Zhu et al. (2021) SCOPE: Flexible targeting and stringent CARF activation enables type III
CRISPR-Cas diagnostics. BioRxiv, https://doi.org/10.1101/2021.02.01.429135.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S115 Unlocking the Power of Next-Generation Sequencing

Alex ARAVANIS
Senior Vice President and Chief Technology Officer, Illumina, USA
Corresponding Author Email : [email protected]

The massively parallel sequencing technology known as next-generation sequencing (NGS) has revolutionized
the biological sciences. With its ultra-high throughput, scalability, and speed, NGS enables researchers to
perform a wide variety of applications and study biological systems at a level never before possible.
Illumina’s Chief Technology Officer, Alex Aravanis PhD, MD, presents on the power and potential of next-
generation sequencing technology and the opportunities to accelerate innovation in the genomics industry in the
clinic and beyond.

Keywords : Illumina, Next Generation Sequencing, Technology

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S116 Multiomics Approach for Targeting Novel Microbiome Therapeutics against Chronic Diseases

GwangPyo KO
Graduate School of Public Health, Seoul National University, Seoul, Korea
Corresponding Author Email : [email protected]

Recently, there were dramatically increased interests on human microbiome research worldwide. While human
ecosystems is maintained through mutualistic relationship between human and microbiota, the effect of gut
microbiota and associated molecular mechanisms on metabolic and immunological diseases has been not much
studied. We have determined the effects of microbiome and determined the genes and pathways of human gut
microbiome using both human populations and animal models. The specific aims of this presentation are 1) to
determine and characterize the composition of human microbiome as related to target chronic diseases, 2) to
investigate the biomarkers for predicting chronic based on microbiome and associated molecular components.
Our data suggest that combination of specific microbiome and associated molecular mechanisms in the gut
predicts the incidences of chronic metabolic diseases. Our research will help us to understand the association
between human microbiome and metabolic diseases and to provide microbiome-based diagnostics and
therapeutics.

References
1. HS Yoon, CH Cho, MS Yun, HJ You, DH Han, KH Cha, SJ J, SH Moon, K Lee, TW Nam, Ko G. 2021.
Akkermansia muciniphila secretes a glucagon-like peptide-1-inducible protein that improve glucose
homeostasis and ameliorate metabolic disease. Nature Microbiol. In press.
2. Lee G, You HJ, Bajai JS, Joo SK, Yu J, Park S, Kang H, Park JH, Kim JH, Lee DH, Lee S, Kim W, and Ko
G. 2020. Distinct signatures of gut microbiome and metabolites associated with significant fibrosis in non-
obese NAFLD. Nature Comm. 10.1038.
3. Seo B, Jeon K, Moon S, Lee K, Kim WK, Jeong H, Cha KH, Lim MY, Kang W, Kweon MN, Sung J, Kim
W, Park JH and Ko G. 2020. Roseburia spp. ameliorate alcoholic fat liver via restoration of gut barrier
functions. Cell Host & Microbe. 8:27(1):25-40.
4. Si J, You HJ, Sung J, and Ko G. 2017. Prevotella as a hub for vaginal microbiota under the influence of host
genetics and their association with obesity. Cell Host & Microbe. 21(1):97-105.
5. Lim MY, You HJ, Yoon HS, Kwon B, Lee KY, Lee S, Song YM, Sung J, Ko G. 2017. The effect of
heritability and host genetics on the gut microbiota and metabolic syndrome. Gut, Apr 6, 2015-311326.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S117 Natural Products Biosynthesis and Their Modification in the Context of Trends in Biocatalysis
and Protein Engineering

Byung-Gee KIM
Seoul National University, School of Chemical and Biological Engineering / Institute of Molecular Biology and
Genetics / Bio-MAX Institute, Seoul, Korea
Corresponding Author Email : [email protected]

In the past 30 years, although many new discoveries and concepts have been developed in the field of
biotechnology, development of biocatalysis and protein engineering was recognized as a process development
of bio-reaction systems and protein catalysts used in the synthesis of fine chemicals in organic chemistry and
bioindustry. As the turn-around time of custom-made biocatalysts development becomes very rapidly shortened
due to variety of technology innovations, and diverse applications of biocatalysts in chemical and bio-industry
are newly opened, biocatalysis already became a good alternative means to substitute chemical catalysis in the
field of organic chemistry, and its coverage and its influence in chemical industry will be rapidly increased and
broadly expanded in the near future.
In this lecture, first, a short review of development of such protein engineering and in vitro evolution will be
presented to understand advances in protein in vitro evolution and mutagenesis during the last 30 years. In the
trends of these protein evolutionary studies, our lab has focused on natural products biosynthesis and their
modifications using the enzymes involved in natural product biosynthesis reactions, such as oxidoreductases
(tyrosinase, reductase, P450 and FAD-dependent monooxygenase), glycosyltransferase and halogenase, and to
overcome any problems and hurdles we have met in the studies, always protein engineering and/or evolution
were required to improve their desired properties, such as activity, enantioselectivity, substrate specificity, and
stability. Here, some examples of related our research activities on synthesis of natural products will be
presented: i) isoflavonoids bioconversion(ortho-hydroxylation, oxygenation, isomerization and reductions), ii)
human milk oligosaccharide production, and iii) indigoids biosynthesis.
Stories of such examples will be in perspectives to evaluate future advances in enzymatic approaches for natural
product biosynthesis.

Keywords : Biocatalysis, Protein engineering, in vitro evolution, natural product synthesis, oxidoreductase,
glycosyltransferase, halogenase
References
1. Jeongchan Lee, Joonwon Kim, Ji Eun Song, Won-Suk Song, Eun-Jung Kim, Yun-Gon Kim, Hee-Jin Jeong,
Hye Rim Kim, Kwon-Young Choi, and Byung-Gee Kim, “Production of Tyrian purple indigoid dye from
tryptophan in Escherichia coli,” (2021) Nature Chemical Biology, 17:104-112.
2. Yun Hee Choi, Bum Seok Park, Joo‐Hyun Seo, Byung‐Gee Kim, “Biosynthesis of the human milk
oligosaccharide 3-fucosyllactose in metabolically engineered Escherichia coli via the salvage pathway
through increasing GTP synthesis and β-galactosidase modification,” (2019) Biotechnology and Bioengi-
neering, 116:3324-3332.
3. Pyung-Gang Lee, Joonwon Kim, Eun-Jung Kim, Sang-Hyuk Lee, Kwon-Young Choi, Romas J.
Kazlauskas, and Byung-Gee Kim, “Biosynthesis of (-)-5-hydroxy-equol and 5-hydroxy-dehydroequol from
soy isoflavone, genistein using microbial whole cell bioconversion”, (2017) ACS Chemical Biology, 12,
2883-2890.
4. Sang-Hyuk Lee, Kiheon Baek, Ju-Eun Lee, Byung-Gee Kim, “Using tyrosinase as a monophenol
monooxygenase: A combined strategy for effective inhibition of melanin formation”, (2016) Biotechnology
and Bioengineering, 113(4):735-743.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S3] 기업특별세션:
Microbiome and
Pre/pro/postbiotics

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S311 Microbiome and Precision Medicine

Hansoo PARK
Department of Biomedical Science and Engineering, Gwangju Institute of Science and Technology,
Gwangju, Korea
Corresponding Author Email : [email protected]

The gut microbiome of patients with disease is believed to influence the development of disease, as well as the
efficacy of the drugs. Human Microbiome research was conducted to understand how microbial communities
impact on human health. However, the genomic characteristics and detailed functions of effective bacterial
strains have not been fully clarified. In this study, we utilized an integrated approach, involving metagenome,
bacterial whole genome/transcriptome, mouse intestinal transcriptome, and mouse serum metabolome analysis,
to decipher whether bacterial strain-specific differences influence the disease susceptibilities and efficacy of
therapeutics in various diseases.

Keywords : Precision Medicine, Microbiome, Immuno-oncology, Obesity

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S312 Microbiome and Probiotics research in Cell Biotech

Sanghyun LIM
Cell Biotech R&D Center, Gimpo, Korea
Corresponding Author Email : [email protected]

Cell Biotech, microbiome and probiotics leading company, was established in 1995. One of specialties in Cell
Biotech is one-stop solution system, which means that we are doing all steps from R&D to sales and education
to keep high-quality of products. In addition, Dual-coating is very-well known technology worldwide.
Probiotics confers a health benefit on host. Many researches have proved the health benefits of probiotics on the
various diseases such as inflammatory disorders and metabolic diseases. Microbiome research is incredibly
increasing with increasing beneficial effects of probiotics on unhealthy subjects. Such research reports that
ingestion of probiotics causes changes in intestinal microbiome, suggesting the possibility of the development of
pharmabiotics for patients. Here I would like to introduce what kinds of research are doing in Cell Biotech.

Keywords : Microbiome, Probiotics, Cell Biotech

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S313 Improve the Quality of Your Life with Human Milk Oligosaccharides

Chulsoo SHIN
Advanced Protein Technologies Corp., Suwon, Korea
Corresponding Author Email : [email protected]

Advanced Protein Technologies Corp. (APTech), is a bio-venture company specialized in developing functional
materials and manufacturing recombinant proteins for biopharmaceutical use with its expertise in microbial
metabolic engineering technology.
In 2016, APTech received the production technology of 2'-Fucosyllactose (2'-FL), a human milk
oligosaccharide available in the highest concentration in human milk, was transferred, from Professor Seo Jin-
ho's team at Seoul National University worked towards commercializing it.
Human breast milk contains full of bioactive components to support the healthy growth and development of
newborn babies in their early stage of life. These bioactive components are consisted of essential microbes,
human milk oligosaccharides (HMOs), immunoglobulins, lactoferrin, cytokines, and fatty acids. HMOs are the
third most abundant solid components after lactose and lipids. HMOs are known over 200 derivatives, combined
with lactose linked to fucose, sialic acids and/or N-acetyl glucosamine. The most abundant HMO is 2’-FL,
which constitutes nearly 30% the total HMOs. Several biological functions have reported the health benefits of
HMOs, which modulation of the microbiota as probiotics, anti-pathogenic effect, anti-viral effect, modulation of
the intestinal epithelial cell response, and development of the immune system.
Recently, APTech received the approval for 2’-FL to be used as a food ingredient in Vietnam, Thailand, and
Malaysia, and US FDA GRAS. Concurrently, 2’-FL is also under review by EFSA for Novel Food approval in
Europe and an application to submit a dossier to China is in preparation. Likewise, food ingredient registrations
for Latin America, Eastern Europe, Japan, and India are also in progress. With 2’-Fucosyllactose as a starting
point, APTech plans to complete a portfolio of the 7 major Human Milk Oligosaccharides including 3-fuco-
syllactose (3-FL), Lacto-N-Tetraose (LNT), Lacto-N-neotetraose (LNnT), Lacto-N-FucoPentaose (LNFP), 3'-
Sialyllatose (3'-SL), and 6'-Sialyllactose (6'-SL). APTech believed that human milk oligosaccharide will be
promising health ingredient and improve the quality of life.

Keywords : 2’-fucosyllactose, Human milk oligosaccharides, food ingredient, Health benefit, APTech

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S314 The Prebiotics, a Mushroom (Schizophyllum commune) Derived β-glucan Effect Beneficially the
Gut Microbiota on Low-fiber Diet or High-fat Diet Mice Model

Sunghong KIM1,2, Moonjae CHO1,2

1
Quegen Biotech, Seoul, Korea, 2Jeju National University, Jeju, Korea
Corresponding Author Email : [email protected]

β-glucan has been studied for various efficacy and can be consumed through grains or mushrooms. But the
intake of β-glucan has not been enough to show its effect. Recently, the development of biotechnology has led
to the continuous efforts to produce pure β-glucan through the cultivation of yeast, bacteria or mushroom
mycelium and introduce it as a health functional food.
In particular, β-glucan has been known to have anti-cancer and anti-inflammatory effects, and recently, interest
in immune function has been increasing due to COVID19 pandemic. In addition, the function as prebiotics is
known along with probiotics, and it is attracting much attention in the development of Synbiotics.
Since 2018, Quegen Biotech has been conducting research on β-glucan intestinal function with Jeju National
University. After completing clinical trials at Jeonbuk National University Hospital last year, it is now analyzing
the usefulness of the human body.
In this study, we present the results of changes in gut microbiota according to the intake of β-glucan in mice fed
HFD (high-fat diet) or LFD(low-fiber diet).

Keywords : Prebiotics, β-glucan, gut microbiota, Synbiotics, high-fat diet mice

References
1. Karthika Muthuramalingam, Moonjae Cho, Dietary intervention using (1,3)/(1,6)-β-glucan, a fungus-
derived soluble prebiotic ameliorates high-fat diet-induced metabolic distress and alters beneficially the gut
microbiota in mic model (2019), European Journal of Nutrition, 59(6), 2617-2629.
2. Karthika Muthuramalingam, Moonjae Cho, Effect of mushroom derived β-glucan on low-fiber diet induced
gut dysbiosis (2019), J Appl Biol Chem, 62(2), 211-217.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

International Program

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S4-1]
Recent Advances in
Genome Editing Technology

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S411 Development of the Base Editing Technology Target-AID and Its Applications

Keiji NISHIDA
Graduate School of Science, Technology and Innovation Kobe University, Japan
Corresponding Author Email : [email protected]

Genome editing is the technology that enables highly efficient and precise modification of genomic DNA
information. Conventional genome editing involves nuclease-mediated DNA double strand breaks at the
targeted site of the genome, which then lead mutagenesis through repair mechanisms of the cell. However, DNA
double strand breaks are toxic to the cell and resultant mutation was random. In place of nuclease activity of
conventional genome editing, newly developed base editing technology employs DNA base-modifying enzymes
to perform direct introduction of point mutations at the target site. Deaminase-mediated base editing (BE and
Target-AID) has been developed by tethering DNA cytidine deaminases to nuclease-deficient CRISPR-Cas9
system, enabling pinpoint C to T mutagenesis within the range of 3-5 bases. It shows much less toxicity
compared to full nuclease Cas9 and does not require template DNA for precise genome editing. It is now
applicable to wide range of organisms and keep improving. Recent developments and applications of the base
editing technology will be discussed.

Keywords : Genome editing, Base editing, Synthetic Biology

References
1. Banno S, Nishida K*, Arazoe T, Mitsunobu H, Kondo A. Deaminase-mediated multiplex genome editing in
Escherichia coli. Nature Microbiolog. Feb 5. doi: 10.1038/s41564-017-0102-6. (2018).
2. Shimatani Z, Kashojiya S, Takayama M, Terada R, Arazoe T, Ishii H, Teramura H, Yamamoto T, Komatsu
H, Miura K, Ezura H*, Nishida K*, Ariizumi T*, Kondo A. Targeted base editing in rice and tomato using a
CRISPR-Cas9 cytidine deaminase fusion. Nature Biotechnology. Mar 27. doi: 10.1038/nbt.3833. (2017).
3. Nishida K, Arazoe T, Yachie N, Banno S, Kakimoto M, Tabata M, Mochizuki M, Miyabe A, Araki M, Hara
KY, Shimatani Z, Kondo A. Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive
immune systems. Science, Aug 4. pii: aaf8729. (2016).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S412 Virus-induced Plant Genome Editing

Sang-Gyu KIM
Department of Biological Sciences, KAIST, Daejeon, Korea
Corresponding Author Email : [email protected]

Plant viruses have been engineered to express heterologous proteins and RNAs in plants for several decades.
This viral system can now be applied to editing plant genomes. Virus vectors can deliver Cas proteins and guide
RNAs, two key components of the CRISPR gene-editing system, into a plant cell without a complicated
experimental procedure. In some cases, plant viruses move to meristematic cells and express gene-editing
components in the cell, which results in the production of mutant seeds. Here, we focus on three main issues of
the virus-induced genome editing (VIGE) technology in plants: (1) how to express the relatively large size of
Cas proteins, (2) how to express guide RNA, and (3) how to increase the efficiency with which viruses are
delivered into meristematic cells. We highlight recent advances in how plant virus vectors can be used
efficiently in plant-genome editing.

Keywords : CRISPR, Plant Genome Editing, Virus

References
1. Youngbin Oh, Hyeonjin Kim, Sang-Gyu Kim, Virus-induced plant genome editing, Current Opinion in
Plant Biology (2021).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S413 Current Status and Challenges of Genome Editing Tools

Sangsu BAE
Department of Chemistry, Hanyang University, Seoul, Korea
Corresponding Author Email : [email protected]

Genome editing tools such as ZFNs, TALENs, and CRISPR-Cas9/Cpf1 derived RNA-guided endonucleases
have been broadly used for biomedical research, biotechnology, and plant transformation. CRISPR nucleases
are widely exploited due to the ease of use and inexpensive cost; researchers can induce gene editing at different
sites by simply altering guide RNAs. Ultimately, the Nobel Prize in Chemistry 2020 were awarded for
discovering one of gene technology's sharpest tools. However, the genome editing tools are constantly being
developed. DNA base editing tools, including cytosine base editors (CBEs) and adenine base editors (ABEs),
enable the direct conversion of DNA bases without producing double-stranded breaks of DNA were developed.
Furthermore, a prime editor (PE) that enables generating small insertion and deletion in addition to substitution
of several nucleotides at target sites, was recently developed. While the gene editing mechanism is different for
each tool, all tools have been developed based on the CRISPR effectors. Here I present current trends in gene
editing tools along with on-going studies of my group such as development of web-based programs in CRISPR
RGEN Tools (www.rgenome.net), protein engineering for enhancing specificity of ABEs, and versatile
application of CRISPR tools for plant transformation and gene editing therapy.

Keywords : CRISPR, Base editing, Prime editing, Gene editing therapy, Web-based programs
References
1. Jeong YK, Lee SH, Hwang G-H, Hong S-A, Park S-e, Kim J-S, Woo J-S* and Bae S*, “Precise adenine
base editors that exhibit minimized cytosine catalysis” (In revision).
2. Kim Y, Yu J, Hong S-A, Eom J, Jang K, Lee S-N, Woo J-S, Jeong J*, Bae S* and Choi D*, “Ex vivo
therapeutic base and prime editing using chemically derived hepatic progenitors in a mouse model of
tyrosinemia type 1” (In revision).
3. Kim C, Jeong YK, Yu J, Shin HJ, Ku KB, Cha HJ, Han JH, Hong S-A, Kim B-T, Kim S-J*, Woo J-S* and
Bae S*, “Efficient Human Cell Coexpression System and Its Application to the Production of Multiple
Coronavirus Antigens,” Advanced Biology (In press). (Featured on the front cover)
4. Jeong YK, Song B and Bae S*, “Current status and challenges of DNA base editing tools” Molecular
Therapy 28, 1938-1952 (2020).
5. Yu J, Cho E, Choi Y-G, Jeong YK, Na Y, Kim J-S, Cho S-R*, Woo J-S* and Bae S*, “Purification of an
intact human protein overexpressed from its endogenous locus via direct genome engineering” ACS
Synthetic Biology 9, 1591-1598 (2020). (Cover article)
6. Kim HS, Jeong YK, Hur JK, Kim J-S* and Bae S*, “Adenine base editors catalyze cytosine conversions in
human cells” Nature Biotechnology 37, 1145-1148 (2019).
7. Jeon Y, Choi YH, Jang Y, Yu J, Goo J, Lee G, Jeong YK, Lee SH, Kim I-S, Kim J-S, Jeong C*, Lee S* and
Bae S*, “Direct observation of DNA target searching and cleavage by CRISPR-Cas12a” Nature Communi-
cations 9, 2777 (2018).
8. Park J, Childs L, Kim D, Hwang G-H, Kim S, Kim S-T, Kim J-S* and Bae S*, “Digenome-seq web tool for
profiling CRISPR specificity” Nature Methods 14, 548-549 (2017).
9. Lim Y, Bak SY, Sung K, Jeong E, Lee SH, Kim J-S, Bae S* and Kim SK*, “Structural roles of guide RNA
in the nuclease activity of Cas9 endonuclease” Nature Communications 7, 13350 (2016).
10. Bae S, Kweon J, Kim HS and Kim J-S, “Microhomology-based choice of Cas9 nuclease target sites” Nature
Methods 11, 705-706 (2014).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S414 Supercharging Engineered T cell Therapies for Cancer with Genome Editing

Jae young LEE

ToolGen Inc., Seoul, Korea
Corresponding Author Email : [email protected]

Immune cell-based therapy, particularly genetically modified T cells such as CAR-engineered T cells has shown
promise in treating certain blood cancers including acute lymphoblastic leukemia and non-Hodgkin lymphomas.
However, clinical utility of CAR-engineered T cells in the context of chronic lymphoblastic leukemia and solid
cancer has been questionable. Efficacious and persistent CAR-T cells should be resistant to tumor-associated
deleterious immunosuppressive molecules such as TGFβ and/or Prostaglandin E2 (PGE2) and effectively
establish long-term memory T cells without exhaustion. Tumor-associated inhibitory signals can dampen the
activity of T cells, impeding the anticancer functions of T cells. Many studies have focused on PD-1 or CTLA-4
blockade to invigorate T-cell functions through CD28 signaling, however obtaining robust clinical outcomes
remains challenging. To overcome inhibitory tumor-associated microenvironment, we propose downregulation
of diacylglycerol kinase (DGK), as it metabolizes diacylglycerol (DAG) to phosphatidic acid (PA) to regulate T
cell function. For this, we utilized CRISPR/Cas9 to knockout of two different isoforms of DGKs (DGKα and
DGKζ) that are expressed in human T cells. Knockout of DGKs augmented the effector functions of CAR-T
cells in vitro via increased TCR signaling. DGKs knockout from CAR-T cells rendered them resistant to soluble
immunosuppressive factors such as TGFβ and PGE2 and sustained effector functions under conditions of
repeated tumor stimulation. Moreover, DGK knockout caused significant regression of several solid tumor
models through enhanced effector functions of either CAR-engineered or TCR-engineered T cells in a xenograft
mouse model. Furthermore, DGK knockout CAR-T cells also showed long-term memory phenotype in
xenograft mouse model even with repeated tumor challenge. Collectively, our study shows that editing of DGKs
effectively enhances the effector functions of CAR-T cells, suggesting that CRISPR/Cas9-mediated knockout of
DGK could be applicable as part of a multifaceted clinical strategy to treat various cancers.

Keywords : CAR-T, TCR-T, CRISPR/Cas9, Cell Therapy, Cancer

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S4-2]
Protein and Enzyme Engineering for
Therapeutic Applications

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S421 Design of Bioconjugates that Function at Biological Interface

Noriho KAMIYA1,2
1
Division of Biotechnology, Center for Future Chemistry, Kyushu University, Japan,
2
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Japan
Corresponding Author Email : [email protected]

By combining the functions of the major components of living organisms, such as proteins, nucleic acids, lipids,
and carbohydrates, it will be possible to design value-added novel bioconjugates. The introduction of synthetic
compounds to naturally occurring components can further expand the utility of the intrinsic biological functions
and even lead to generate new functions. There is growing interest in the use of enzymes that catalyze site-
specific crosslinking reactions for this purpose in order to minimize the impact of chemical modification on the
structure and function of biomolecules upon bioconjugation. In this context, we have demonstrated the potential
of oxidoreductases such as horseradish peroxidase (HRP)1 and laccase2 for crosslinking reaction that is specific
to a tyrosine residue genetically introduced into the protein of interest. Microbial transglutaminase (MTG) has
also been widely used to crosslink a variety of functional molecules such as biotin3, nucleic acids4, synthetic
polymers5, lipids6 and drugs7 with proteins. In this presentation, our on-going efforts to create novel biocon-
jugates that function at the biological interface will be introduced.

Keywords : Enzymatic crosslinking, Horseradish peroxidase Microbial transglutaminase, Laccase, Lipidization

References
1. K. Minamihata, M. Goto, N. Kamiya, Site-specific conjugation of an antibody binding protein catalyzed by
horseradish peroxidase creates a multivalent protein conjugate with high affinity to IgG. (2015) Biotechnol.
J., 10, 222-226.
2. D. Permana, K. Minamihata, R. Sato, R. Wakabayashi, M. Goto, N. Kamiya, Linear polymerization of
protein by sterically-controlled enzymatic cross-linking with a tyrosine-containing peptide loop. (2020)
ACS Omega., 5, 5160-5169.
3. Y. Mori, H. Nakazawa, G.A.L. Goncalves, T. Tanaka, M. Umetsu, N. Kamiya, One-dimensional assembly
of functional proteins: toward the design of an artificial cellulosome. (2016) Mol. Syst. Des. Eng., 1, 66-73.
4. M. Takahara, R. Wakabayashi, K. Minamihata, M. Goto, N. Kamiya, Primary amine-clustered DNA
aptamer for DNA-protein conjugation catalyzed by microbial transglutaminase. (2017) Bioconjugate
Chem., 28, 2954-2961.
5. R. Wakabayashi, K. Yahiro, K. Hayashi, M. Goto, N. Kamiya, Protein-grafted polymers prepared through a
site-specific conjugation by microbial transglutaminase for an immunosorbent assay. (2017) Biomacro-
molecules, 18, 422-430.
6. [1] M. Takahara, R. Wakabayashi, N. Fujimoto, K. Minamihata, M. Goto, N. Kamiya, Enzymatic cell-
surface decoration with proteins using amphiphilic lipid-fused peptide substrates. (2019) Chem. Eur. J., 25,
7315–7321.; [2] M. Takahara, S. Mochizuki, R. Wakabayashi, K. Minamihata, M. Goto, K. Sakurai, N.
Kamiya, Extending the half-life of a protein in vivo by enzymatic labeling with amphiphilic lipopeptides.
(2021) Bioconjugate Chem. DOI: 10.1021/acs.bioconjchem.1c00027.
7. P. Strop, Versatility of Microbial Transglutaminase (2014) Bioconjugate Chem., 25, 855-862.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S422 Repurposing Biology through Synthetic Enzymology

Wen Shan YEW

Department of Biochemistry, Yong Loo Lin School of Medicine, and Synthetic Biology for Clinical and Techno-
logical Innovation, National University of Singapore, Singapore
Corresponding Author Email : [email protected]

Synthetic Enzymology has been increasingly used to engineer enzymes within biological systems for the
repurposing of biology for purposeful function. On that note, the talk will touch on the Synthetic Cannabinoid
Biology Programme (SCBP) that is part of the National Research Foundation’s (NRF) Synthetic Biology R&D
Programme. The SCBP aims to discover new biosynthetic pathways that produce cannabinoid compounds in a
sustainable manner. The SCBP does not study the psychoactive compounds of the cannabis plant, such as
tetrahydrocannabinol (THC), which is illegal in Singapore. Instead, it identifies cannabinoid genes for the
sustainable production of cannabinoids, such as cannabidiol (CBD). The SCBP does not use the cannabis plant
as the aim is to produce non-psychoactive cannabinoids through synthetic biology, so that benefits of
cannabinoid pharmaceuticals can be reaped without the use of the plant. In this talk, we will highlight some
recent efforts in the sustainable production of cannabinoids using synthetic biology. In addition, the use of
mechanistic and engineering principles in enzymology will also be highlighted to demonstrate the utility of
synthetic enzymology in a range of applications from therapeutics to sustainable manufacturing to bioreme-
diation.

Keywords : Synthetic Cannabinoid Biology, Synthetic Enzymology, Synthetic Biology

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S423 Discovery and Engineering of Human Antibodies Against G Protein-Coupled Receptors

Sang Taek JUNG

Department of Biomedical Sciences, Korea University, Seoul, Korea
Corresponding Author Email : [email protected]

Antibody molecules have been widely used for therapeutics, diagnostics, and biomedical research reagents due
to their outstanding target specificity and evolvability. For efficient isolation of monoclonal antibodies against
highly valuable targets (e.g., soluble oncogenic targets, receptors, toxins, etc.) in a high throughput manner, we
have constructed a series of human antibody libraries, which have a vast diversity of over 1× 1011, and have
successfully utilized them for the isolation of monoclonal antibodies against highly challenging target antigens
including G protein-coupled receptors (GPCRs). GPCR plays pivotal roles in proliferation and metastasis of
many types of cancer, and its expression is highly correlated with prognosis and survival rate of cancer patients.
In this study, we have successfully prepared endothelin receptor type A antigen, and screening of our in-house
human antibody library could generate anti-ETA antibodies antagonizing the ETA-mediated cellular signaling.
In vitro and in vivo anti-tumor potency of the resulting antibodies have been thoroughly evaluated and the
results will be presented.

Keywords : Antibody Engineering, G Protein-Coupled Receptors, Fc Engineering

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S424 Multivalent Albumin-FcRn Interactions for the Significantly Prolonged Serum Half-life of a
Therapeutic Protein

Inchan KWON
School of Materials Science and Engineering, Gwangju Institute of Science and Technology (GIST), Gwangju,
Korea
Corresponding Author Email : [email protected]

Human serum albumin (HSA) has been investigated as a serum half-time extender of therapeutic proteins owing
to its exceptionally long serum half-life via the neonatal Fc receptor (FcRn)-mediated recycling mechanism. In
most cases, only one HSA molecule was conjugated to a therapeutic protein leading to a limited extension of
serum half-life. We hypothesized that conjugation of multiple HSA molecules to a therapeutic protein
significantly further extends the serum half-life via multivalent HSA-FcRn interactions. We chose urate oxidase
(Uox), a tetrameric therapeutic enzyme used for the treatment of gout, as a model. In previous studies, only one
HSA molecule was site-specifically conjugated to one Uox because of poor conjugation yield of the relatively
slow bioorthogonal chemistry, strain promoted azide-alkyne cycloaddition. To increase the number of HSA
molecules conjugated to one Uox, we employed the faster bioorthogonal chemistry. We prepared Uox variants
conjugated to one, two, or four HSA molecules. The enzyme activity of all three Uox-HSA conjugates was
comparable to that of unmodified Uox. We found out that an increase in HSA molecules conjugated to Uox
(multiple albumin-conjugated therapeutic protein) enhanced FcRn binding and consequently prolonged the
serum half-life in vivo.

Keywords : FcRn-mediated recycling, half-life extension, serum albumin, site-specific albumin conjugation,
urate oxidase, bioorthogonal chemistry

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S425 Engineering Antibodies for Superior Pharmacokinetic Properties

Tae Hyun KANG

School of Applied Chemistry, Kookmin University, Seoul, Korea
Corresponding Author Email : [email protected]

Antibodies are a cornerstone molecule in humoral immunity. Fc domain of antibodies takes part in two major
functions, which are i) effector functions of immune effector cells and ii) serum half-life of IgG antibodies via
Fc receptors (FcRs) binding. Thus, engineering Fc towards selective FcRs affinity will lead to antibodies with
guided effector functions or with superior pharmacokinetic properties.
In this seminar I am going to talk about engineering antibody Fc for increasing serum half-life, capitalizing on
sophisticated directed evolution strategies. Therapeutic potencies of the biochemically evolved Fc’s are
validated using human neonatal Fc receptor (hFcRn)-transgenic mice in vivo.

Keywords : Antibody Engineering, Fc receptors (FcRs), Pharmacokinetics

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S4-3]
Trends in Microbiome Engineering

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S431 Engineering Microbes to Rewire Host-microbiome Interactions

Matthew CHANG
NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), and Synthetic Biology
Translational Research Program and Department of Biochemistry, Yong Loo Lin School of Medicine, National
University of Singapore, Singapore
Corresponding Author Email : [email protected]

The wealth of knowledge on the human microbiota composition and its roles in health and disease has recently
spurred the development of novel therapeutic strategies. Moreover, with an array of genetic tools that are readily
available, programmable genetic circuits can be designed, genomes can be edited and rewritten, and cells can be
reprogrammed to foster novel microbiota-based interventions. In this talk, our recent work on engineering gut-
resident microbes as versatile platforms equipped with clinically relevant functionalities will be presented. A
particular emphasis will be placed on our efforts to transform gut microbes into live biotherapeutics with
prophylactic and therapeutic efficacy against pathogenic infections and chronic metabolic diseases. This work
provides a strong foundation for engineering microbes to modulate host-microbiome interactions and supports
the use of live biotherapeutics as a viable strategy for clinical intervention.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S432 Genome-wide Multi-omics Analysis of Mucin-degrading Bacteria Reveals Their Nutrient-

dependent Distinct Metabolic Features in the Human Gut

Kyoung Su KIM, Dong-Woo LEE

Department of Biotechnology, Yonsei University, Seoul, Korea
Corresponding Author Email : [email protected]

Diet-driven gut microbiota is highly associated with human health and diseases. Mucin-degrading (MD) bacteria
play a crucial role in shaping commensal microbiota's structure and function in the human gut, but their
metabolic features remain elusive. Herein we performed a genome-wide analysis to investigate the nutrient-
dependent physiology and metabolism of MD bacteria. The genome sequences of Ruminococcus gnavus ATCC
29149 (RG, 3.55 Mb) and Akkermancia muciniphila ATCC BAA-835 (AM, 2.66 Mb) reveal that they contained
3,345 and 2246 protein-encoding genes, respectively, according to the NCBI Prokaryotic Genome Annotation
Pipeline (PGAP). However, the PGAP annotations exhibited >60% mismatches with those from different
pipeline configurations. Accordingly, we revised the genome annotations of both MD bacteria using the
UniProtKB/Swiss-Prot database, followed by additional manual curation based on sequence and structural
domain similarity searches. Consequently, we revised 25 genes for RG and 47 genes for AM involved in mucin-
derived fermentation pathways and their distinct MD enzymes. To validate the reconstructed pathways, we
investigated MD bacteria's nutrient-dependent physiological features and their fermentative profiles using
comparative transcriptomic analysis. As a result, we identified their distinct propionate fermentation pathways
together with mucin-derived microbial metabolites such as indole-3-lactate and leucic acid by LC-MS/MS
spectrometry. Our multi-omics analyses integrated with physiological characterization under anaerobic
conditions reveal MD bacteria's differential metabolic features, providing insight into their metabolic
consequences of nutrient-dependent phenotypes on microbiota networks with commensal bacteria in the human
gut.

Keywords : Mucin-degrading bacteria, genome annotation, anaerobic fermentation, Ruminococcus gnavus,

Akkermansia muciniphila

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S433 Strict Anaerobic Microbiota to Clinical Application

Byoung-Chan KIM
Metabolic Regulation Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon,
Korea / 114 Bioventure Center, HealthBiome, Inc., Daejeon, Korea
Corresponding Author Email : [email protected]

The study of the human microbiome has come to the forefront of biological research. The human symbiotic
microbiota and its metabolites influence not only gastrointestinal and nutritional diseases such as Clostridium
difficile infection (CDI), irritable bowel syndrome, inflammatory bowel disease, obesity and diabetes but also a
host of other human diseases including cancer, arteriosclerosis, Alzheimer’s (AD), and Parkinson’s (PD)
disease. However, most research into the human microbiome thus far has focused on microbial diversity and
metagenomic analysis through next-generation sequencing (pyrosequencing) techniques. In the next phase,
research should focus on the isolation of foundation species and their clinical applications for preventing and
curing these microbiota-related human diseases. Recent research has shown that obligate anaerobes occupy
more than 99% of total gut microbes and they have a more profound impact on human diseases than facultative
ones. Several global drug companies have created cocktails of human gut strict anaerobes for use in faecal
microbiota transplantation (FMT); some of these cocktails are currently undergoing clinical trials as treatments
for human diseases, and are almost ready for commercial release. Here, I will show one example of the
importance of strict anaerobe for regulating neuro-related human disease such as AD, PD, autism spectrum
disorder (ASD) and depression. Agathobaculum butyriciproducens (Abut) is a strict anaerobic and butyric acid-
producing bacteria. Abut administration to AD, PD, ASD and depression mouse models, improved the
phenotypic mouse behaviours and relieve the relative disease symptoms. In conclusion, Abut can be a candidate
of Live Bio therapeutic Product (LBP) for preventing or even curing human neuro-related diseases.

Keywords : Microbiome, Strict Anaerobes, Agathobaculum, LBP

References
1. Go et al., Human gut microbiota Agathobaculum butyriciproducens improves cognitive impairment in LPS-
induced and APP/PS1 mouse models of Alzheimer's disease (2021), Nutrition research, 86, 96-108.
2. Eckburg et al., Diversity of the Human Intestinal Microbial Flora (2005), Science, 308, 1635-1638.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S434 Engineering Genetic Circuits for Sensing Gut Inflammation

Dae-Hee LEE
Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea
Corresponding Author Email : [email protected]

Trillions of microbes are living and interacting with host cells in the human gut. Since they are essential to
human health, changes in the composition of this gut microbiome cause disease-associated symptoms, which
motivates the engineering of microbes to sense, treat, or prevent disease. Microbes reprogrammed using
advanced genetic circuits are envisaged as emerging living diagnostics for a wide range of diseases and play key
roles in regulating gut microbiota to treat disease in a non-invasive manner. Here, we developed a smart
Escherichia coli that senses and responds to nitrate, a biomarker of gut inflammation.
To this end, we first employed the NarX-NarL two-component regulatory system in E. coli to construct a
nitrate-responsive genetic circuit. Next, we optimized this genetic circuit for the best performance using
measures of sensitivity and specificity. We then introduced the nitrate genetic circuit into a well-known
probiotic E. coli Nissle 1917. We demonstrated that the designed genetic circuit could sense elevated nitrate
levels during gut inflammation in mice with native gut microbiota. Moreover, using Boolean AND gate, we
generated a genetic circuit for simultaneous sensing of the thiosulfate and nitrate biomarkers, thus increasing the
tool’s specificity for diagnosing gut inflammation. The nitrate-responsive genetic circuit will enable new
approaches for non-invasive diagnostics of inflammation-associated diseases.

Keywords : Genetic circuit, Gut inflammation, Designer probiotic, Nitrate sensing, Thiosulfate sensing,
Boolean AND gate
References
1. Seung-Gyun Woo et al., A designed whole-cell biosensor for live diagnosis of gut inflammation through
nitrate sensing (2020), Biosensors and Bioelectronics, 168, 112523.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S4-4]
Recent Progress of
Bio-based Plastics

31
2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S441 Closing Cycles in the Plastics Bioeconomy Using Extremophilic Bacteria

Nick WIERCKX
Jülich Research Centre, Institute for Bio- and Geosciences IBG-1:Biotechnology, Jülich, Germany
Corresponding Author Email : [email protected]

The environmental impact of fossil-based plastics has been broadly discussed. The carbon footprint of
traditional plastics contributes to climate change, and especially for more complex polymers or mixed waste
stream, technical recycling options are still extremely limited.
On the one hand, we produce bio-based plastic monomers using the extremely solvent-tolerant Pseudomonas
taiwanensis. This strain was engineered to produce drop-in materials such as phenol, 4-hydroxybenzoate, and
styrene from glucose or glycerol (1). These developments can enable the production of novel and more
sustainable polymers, but they don’t necessarily solve the problem of end-of-life plastic pollution.
Thus, on the other hand, we work to establish plastic waste as a novel bulk carbon source for industrial
biotechnology (2). We have engineered Pseudomonas to metabolize ethylene glycol (3), 1,4-butanediol (4), and
adipic acid as aliphatic monomers of many polyesters and polyurethanes. The resulting strains were genomically
and biochemically characterized. The gained insights enable engineering of plastic monomer metabolism in
different Pseudomonas strains that produce various value-added products.
By tackling both the bio-based production of plastics and the biodegradation of plastics we can create new value
cycles that incentivize waste collection and therefore reduce the burden on the environment.

Keywords : Plastic hydrolysates, plastic monomers, upcycling, Pseudomonas

References
1. T. Schwanemann, M. Otto, N. Wierckx, B. Wynands. 2020. Pseudomonas as versatile aromatics cell
factory. Biotechnol. J. 15:1900569.
2. N. Wierckx, M. Auxiliadora Prieto, P. Pomposiello, V. de Lorenzo, K. O’Connor, L.M. Blank. 2015. Plastic
waste as a novel substrate for Industrial Biotechnology. Microb. Biotechnol. 8:900-903.
3. W.J. Li, L.N. Jayakody, M.A. Franden, M. Wehrmann, T. Daun, B. Hauer, L.M. Blank, G.T. Beckham, J.
Klebensberger, N. Wierckx. 2019. Laboratory evolution reveals the metabolic and regulatory basis of
ethylene glycol metabolism by Pseudomonas putida KT2440. Environ. Microbiol. 21:3669-3682.
4. W.J. Li, T. Narancic, S.T. Kenny, P.-J. Niehoff, K. E. O'Connor, L.M. Blank, N. Wierckx. 2020. Unraveling
1,4-butanediol metabolism in Pseudomonas putida KT2440. Front. Microbiol. 11:382.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S442 Utilizing Formate for Biosynthesis of Polyhydorxyalkanoate (PHA)

Jihee YOON, Min-Kyu OH

Department of Chemical and Biological Engineering, Korea University, Seoul, Korea
Corresponding Author Email : [email protected]

Formate is a promising and sustainable feedstock synthesized from carbon monoxide (CO) or carbon dioxide.
Methylorubrum extorquens AM1 is a type II methylotroph that can use formate as a sole carbon source. In this
study, we engineered this strain to convert formate into poly-3-hydroxybutyrate (PHB), a promising degradable
biopolymer. First, we developed two-stage bioreactor. Acetobacterium woodii was used to transform CO to
formate with a whole cell catalysis first-stage reactor. When the strain’s energy metabolism was blocked to
suppress acetate production, formate was produced from CO with yield of 100%. The resulting formate was
separated and fed to a second-stage reactor. In this bioreactor, an engineered Methylbacterium extorquens AM1
was used for biopolymer synthesis. With optimized pH, formate uptake, and biopolymer synthetic pathway, a
high rate production of PHB was conducted. Although PHB production from CO was realized by two stage
bioprocess, mechanical property of PHB is very poor to substitute plastics. Therefore, production of PHA
copolymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), is desired. For this purpose, the endo-
genous PhaA and PhaC were replaced by bktB and phaC2, with broader substrate specificities. Also, PhaJ was
additionally expressed. The engineered M. extorquens produced PHBV with 8.9% 3HV using formate as the
sole carbon source. In addition, 3HV portions was increased up to 70.6% when cheap SCFA’s were supplied.
This study showed that a PHBV can be produced from CO through metabolic engineering and controlled
bioprocesses.

Keywords : Biopolymer, Formate, Polyhydorxyalkanoate (PHA), Metabolic engineering, Carbon monoxide

References
1. H.W. Hwang, J. Yoon, K. Min, M.S. Kim, S.J. Kim, D.H. Cho, H. Susila, J.G. Na, M.K. Oh, Y.H. Kim,
Chem. Eng. J. 389:124394 (2020).
2. J. Yoon, W. Chang, S.H. Oh, S.H. Choi, Y.H. Yang, M.K. Oh, Int. J. Biol. Macromol. 177:284-293 (2021).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S443 Development of Biodegradable Plastics Using Bio Based Chemicals

Hoon RYU
Industrial Biotechnology PU, Samyang Corp., Seoul, Korea
Corresponding Author Email : [email protected]

Global warming and waste plastics issues are recognized as important environmental issues. One of important
materials to cope with these issues is eco-friendly plastics.
Eco-friendly plastics can be divided into non-degradable bioplastics and biodegradable plastics. Non-degradable
bioplastics which use biomass derived chemicals play an important role in global carbon neutrality. Bio-
degradable plastics are mainly produced using monomers derived from biomass or fermentation product.
Biodegradable plastics using bio based chemicals are very important in terms of global warming delay by
reducing carbon dioxide in the atmosphere and decrease in waste plastic issues.
In this presentation, the development and applications of biodegradable plastics using bio based chemical
materials such as isosorbide along with the market prospect of eco-friendly plastics will be introduced.

Keywords : Bio based chemical, biodegradable plastics, Global warming, Waste plastic, Isosorbide

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S444 Chemo- and Bio-cascade Catalysis to Produce Plastic Monomers from Polyethylene Terephthalate

Jeong Chan JOO1, Si Jae PARK2, Kyoung Heon KIM3, Dongyeop OH4, Bong Keun SONG4, Hyun Gil CHA4,
Bong Hyun SUNG5, Young Joo YEON6, Hee Taek KIM7
1
Department of Biotechnology, The Catholic University of Korea, Bucheon, Korea, 2Division of Chemical
Engineering and Materials Science, Ewha Womans University, Seoul, Korea, 3Department of Biotechnology,
Graduate School, Korea University, Seoul, Korea, 4Research Center for Bio-based Chemicals, Korea Research
Institute of Chemical Technology, Ulsan, Korea, 5Synthetic Biology and Bioengineering Research Center, Korea
Research Institute of Bioscience and Biotechnology, Daejeon, Korea, 6Department of Biochemical Engineering,
Gangneung-Wonju National University, Gangneung, Korea, 7Department of Food Science and Technology,
College of Agriculture and Life Sciences, Chungnam National University, Daejeon, Korea
Corresponding Author Email : [email protected]

Biological depolymerization of polyethylene terephthalate into monomers has gained great attention to reutilize
PET waste as renewable materials. Recently, PETases, MHEtases, and cutinases have been discovered and
engineered to efficiently degrade PET into monomers for regeneration PET. However, these PET-hydrolytic
enzymes still have some limitations in PET depolymerization such as low activity and solubility. Chemo- and
bio-cascade depolymerization of PET can be used to overcome limitations of biological depolymerization. In
this talk, combination of chemical catalyst and biocatalyst will be presented for the upcycling of PET into value-
added chemicals such as plastic monomers or coating agent.

Keywords : Chemo- and bio catalysis, cascade reaction, biological upcycling, PET (polyethylene terephthalate)
References
1. Kang M.J., Kim H.T., Lee M., Kim K.A., Khang T.U., Song H.M., Park S.J., Joo J.C., and Cha H.G. Green
Chemistry 22 (11), 3461-3469 (2020).
2. Kim H.T., Kim J.K., Cha H.G., Kang M.J., Lee H.S., Khang T.U., Yun E.J., Lee D.-H., B.K. Song, Park
S.J., Joo J.C., and Kim K.H. ACS Sustainable Chemistry & Engineering, 7 (24), 19396-1940 (2019).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S4-5]
BEST-KSBB Joint Symposium

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S451 The Challenges and Opportunities of Synthetic Biology

I-Son Grace NG
Department of Chemical Engineering, National Cheng Kung University, Tainan, Taiwan
Corresponding Author Email : [email protected]

In the past seven years, I initiated my “Functional Genes and Proteomics Laboratory” and “iGEM_NCKU_
Tainan for undergraduate students” in National Cheng Kung University. Our mission is to solve the problems by
using technologies and concepts of synthetic biology for the future. In this talk, I would introduce our research
interests which including: (1) Chemical production of 5-aminolevulinic acid by “AI Cell”, which we engineered
a new strain of Escherichia coli, called PIECE. In this strain, the super salvage pathway was fine-tuned to
enhance the regeneration of pyridoxal 5’-phosphate (PLP), on which the activity of ALA synthase (ALAS) is
highly dependent. The metabolic flux analysis indicated that PIECE would redirect more carbon into the
tricarboxylic acid (TCA) cycle and generate more glycine and glutamate, thereby increasing the ALA and
biomass. Finally, the maximum ALA yield of 8.21 g/L and a productivity of 0.228 g/L/h were obtained. The
results showed that PIECE was not only feasible for high-level ALA production but also had significant
potential to reprogram the carbon flux in the TCA cycle to produce value-added chemicals. (2) We developed
the “whole-cell biosensor for diagnostic and prevention” for diabetes. A synthetic cell which coupling plasmid-
driven T7RNA polymerase (PDT7) and NIMPLY-mediated CRISPRi would increase the sensitivity for
diagnostic and finally was applied to an integrative and portable system for daily urine glucose inspection. (3)
We used CRISPR/Cas9 system to enhance lipid accumulation in microalgae, Chlorella vulgaris. the
Agrobacterium-mediated plasmid is suitable for gene insertion in Chlorella species. The left-border and right-
border assisted Cas9 with sgRNA designed on omega-3 fatty acid desaturase (fad3) gene to obtain a higher
accumulation of lipid content by 46% (w/w) in C. vulgaris FSP-E. This is first-time to use CRISPR/Cas9 based
technology for gene manipulation in Chlorella. Finally, I would share 4 projects done by iGEM_NCKU_Tainan
from 2016 to 2019, especially for the project of “Oh My Gut” which we got the Undergraduate Grand Prize
Winner in 2019. No matter what kinds of challenges we will face, synthetic biology is one of the attractive
opportunities to the next generation.

Keywords : Synthetic biology, 5-aminolevulinic acid, biosensor, CRISPR/Cas9, microalgae, iGEM

References
1. Chengfeng Xue, Tzu-Hsuan Yu, I-Son Ng, Engineering pyridoxal kinase PdxY-integrated Escherichia coli
strain and optimization for high-level 5-aminolevulinic acid production. Journal of the Taiwan Institute of
Chemical Engineers, (2021) in press.
2. Shih-I Tan, I-Son Ng, CRISPRi-mediated NIMPLY logic gate for fine-tuning the whole-cell sensing toward
simple urine glucose detection. ACS Synthetic Biology, (2021) 10(2), 412-421.
3. Way-Rong Lin, I-Son Ng, Development of CRISPR/Cas9 system in Chlorella vulgaris FSP-E to enhance
lipid accumulation. Enzyme and Microbial Technology, (2020) 133: 109458.
4. Way-Rong Lin, Shih-I Tan, Chuan-Chieh Hsiang, Po-Kuei Sung, I-Son Ng, Challenges and opportunity of
recent genome editing and multi-omics in cyanobacteria and microalgae for bio-refinery. Bioresource
Technology, (2019) 291, 121932.
5. https://2019.igem.org/Team:NCKU_Tainan

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S452 On-site pre-detection of pathogens using colorimetric DNA biosensors

Byeong Hee HWANG

Division of Bioengineering, Incheon National University, Incheon, Korea
Corresponding Author Email : [email protected]

On-site pre-detection of pathogens including COVID-19 could significantly decrease of a disease outbreak or
national loss in most of the countries. However, conventional detection techniques were limited in use for on-
site detection due to the necessity of specialized skill or equipment. Therefore, it is necessary to develop a new
technique that can pre-detect pathogens in the field without special skills or equipment. Here, we presented a
novel genetic biosensor based on enzyme-free dual-amplification of universal hybridization chain reaction
(uHCR) and hemin/G-quadruplex horseradish peroxidase (HRP)-mimicking DNAzyme and a DNAzyme
strategy to control a plasmonic biosensor for rapid and simple visual detection of Salmonella Choleraesuis. As a
result, the proposed biosensors could visually detect single nucleotide polymorphisms or pathogens without
unstable enzymes, a specialized technique, or equipment. Therefore, these advantages could allow that this
biosensor would be used for on-site pre-detection to lower the risk of transmission of infectious diseases
including COVID-19.

Keywords : On-site detection, Colorimetric, DNA Biosensor, Pathogens

38
2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S453 Programming Cells for Detecting and Degrading Phenolic Compounds

Shen-Long TSAI
Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan

With industrialization and urbanization, our daily life has become much more convenient than ever. However,
problems associated with such modernization are gradually appearing. Especially, environmental pollution is
one of the most major problems people are facing now. 1 Therefore, our greatest challenge is to find means to
deal with the issue. In my lab, we have employed genetic engineering techniques to address these issues. In this
talk, I am going to demonstrate how we can reprogram a bacterial strain to sense, seek and destroy phenolic
compounds. Via harnessing the transcription factors, 2 we systematically created several biological crafts to
detect phenols. To control the cell movement, the chemotaxis system was analyzed and remodeled. 3 Finally, a
phenol utilization pathway was appended for the degradation of phenol.

Keywords : Phenol, biodegradation, detection, chemotaxis

References
1. Maurya, D. K., Kumar, A., Chaurasiya, U., Hussain, T., Singh, S. K. Modern era of microbial biotechno-
logy: opportunities and future prospects. (2021) Microbiomes and Plant Health, 317-343.
2. Mitchler, M. M., Garcia, J. M., Montero, N. E., Williams, G. J. Transcription factor-based biosensors: a
molecular-guided approach for natural product engineering. (2021) Current Opinion in Biotechnology, 69,
172-181.
3. Arumugam, G., Tyagi, J. Keller-Segel chemotaxis models: a review. (2021) Acta Applicandae Mathe-
maticae, 171(1), 1-82.
4. Zhou, Weiwei, et al. Metabolomics analysis reveals potential mechanisms of phenolic accumulation in
lettuce (Lactuca sativa L.) induced by low nitrogen supply. (2021) Plant Physiology and Biochemistry 158,
446-453.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S454 Mechanical Force-Responsive Microcapsules for Self-Regulated Therapeutics Release in

Dynamically Loaded Environments

Yun Kee JO
Department of Biomedical Convergence Science and Technology, Kyungpook National University,
Daegu, Korea
Corresponding Author Email : [email protected]

Bacterial infection of a wound is a major complication that can significantly delay proper healing and even
necessitate surgical debridement and other complications. Conventional non-woven fabric dressings, including
gauzes, bandages and cotton wools, often fail in treating wound infections in a timely manner due to their
passive release mechanism of antibiotics. Here, we propose adhesive mechanically-activated microcapsules
(MAMCs) capable of strongly adhering to a fibrous matrix to achieve a self-regulated release of antibiotics upon
uniaxial stretching of a non-woven fabric dressings. To achieve this, a uniform population of polydopamine
(PDA)-coated MAMCs (PDA-MAMCs) are prepared using a microfluidics technique and subsequent oxidative
dopamine polymerization. The PDA-MAMC allows for robust mechano-activation within the fibrous network
through high retention and effective transmission of mechanical force under stretching. By validating the
potential of a PDA-MAMC-laden gauze to release antibiotics in a tensile strain-dependent manner, we
demonstrate that PDA-MAMCs can be successfully incorporated into a woven material and create a smart
wound dressing for control of bacterial infections. This new mechano-activatable delivery approaches will open
up a new avenue for a stretch-triggered, on-demand release of therapeutic cargos in skin-mountable or wearable
biomedical devices.

Keywords : stretch-responsive, mechano-activation, adhesive microcapsules, fabric wound dressing, antibiotics

delivery
References
1. Y. K. Jo, D. Lee, Small 1903736 (2020).
2. A. P. Perodo, Y. K. Jo, G. Duan, G. R. Dodge, D. Lee, R. L. Mauck, Biomaterials 265, 120255 (2021).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S4-6]
Emerging NGS Tools for
Industrial Biotechnology

41
2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S461 FACS-seq as a Powerful Tool for Profiling the Dose-response Curves of Biosensors in a Massively
Parallel Manner

Chong ZHANG
Department of Chemical Engineering, Tsinghua University, Beijing, China
Corresponding Author Email : [email protected]

Living organisms have a variety of mechanisms to sense and respond to the environmental signals by dyna-
mically regulating their genetic expression networks. Harnessing this ability, genetically encoded biosensors,
mimicking natural regulation networks by assembling basic biological parts like promoter, ribosome binding
site, operator, reporter etc. into genetic circuit, are developed to recognize the analytes and transduce their
signals to a measurable output. Dose-response curve, mapping the genetic circuits to their function, shapes how
individual biosensors respond to the specific signals, which is crucial not only for the specific usage scenario of
genetic encoded biosensor, but also for illustrating their regulatory functions in living cells. In this report, we
would like show our recent attempts on the application of a dose-response profiling method, namely
fluorescence-activated cell sorting coupled with next-generation sequencing (FACS-seq), in generating accurate
dose-response curves for thousands of biosensors in a massively parallel manner, which provides a powerful
platform for dissecting the mechanistic basis of the regulatory elements in living cells, and for the fine tuning of
biosensors in a customized and low-cost manner.
As the first example, we focused on tnaC, which encodes the tryptophan operon leader peptide in bacteria and is
a model of macromolecular-machinery-dynamics-dependent regulatory elements. Working as a molecular
sensor, tnaC responds to intracellular tryptophan (Trp) and regulates the biosynthesis of indol. We used FACS-
seq to generate accurate response curves for all possible codon substitutions in tnaC. The FACS-seq results
allowed us to generate comprehensive genotype-phenotype maps for 1,450 tnaC variants in living cells. The
results clarified the nature of several transient, previously enigmatic states involving RNAP and the ribosome,
and these states play important roles in the tnaC sensor response. Using in silico modeling and additional
experiment, we further demonstrated the molecular basis of the quantitative relation between basal and inductive
response, as well as the range of detection of the sensor.
In the second example, we developed a novel biological parts assembly workflow to encode genetic circuits
with short DNA barcodes, which make sure one-to-one correspondence of the barcodes and biological parts
combinations, enabling high-throughput generation of dose-response curves of higher-order combinatorial
biosensor space. As a proof of concept, we applied our novel workflow for the fine-tuning of the dose-response
curve of malonyl-CoA biosensor based on FapR-fapO system in Saccharomyces cerevisiae. With our 5-step
encoding workflow, a trackable combinatorial library contained 5155/5184 combinations with 6 levels of TF
dosage, 4 different operator positions, and 216 possible UAS designs, was constructed. FACS-seq successfully
characterized the response curve of 2,632 biosensors out of 5184 combinations, providing large-scale genotype-
phenotype association data of the designed biosensors. Machine-learning algorithms were then developed to
predict uncharacterized dose-response curves and identify key features in the whole combinatorial library,
generating a panoramic scanning map of the combinatorial space. With the assistance of our novel workflow,
3755 dose-response curves were obtained at a cost of $1.37 per curve, and a malonyl-CoA biosensor with the
largest dynamic response range reported so far was successfully acquired.

Keywords : FACS-seq, biosensor, dose-response curve, tryptophan, malonyl-CoA

References
1. Wang, T.*, Zheng, X., Ji, H., Wang, T. L., Xing, X. H., & Zhang, C.*. Dynamics of transcription-
translation coordination tune bacterial indole signaling. Nature Chemical Biology 16, 440-449 (2020).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S462 The Fur Pan-regulon Uncovers the Complexity and Diversity of Transcriptional Regulation in
Escherichia coli

Donghyuk KIM
School of Energy and Chemical Engineering, Ulsan National Institute of Science and Technology, Ulsan, Korea
Corresponding Author Email : [email protected]

Global transcription factors (TFs) have been studied extensively in the reference strain Escherichia coli K-12
MG1655 for the past few decades. However, conservation and possible diversity of regulons has not been
experimentally investigated. With experimentally-derived pan-regulon definition for the highly conserved global
Ferric uptake regulator (Fur), involved in iron homeostasis, we determined Fur regulons in nine E. coli strains of
different phylogroups with ChIP-exo and RNA-seq. Thirty-six of the 469 target genes (8%) found belong to the
core regulon, comprising genes involved in ion transport/metabolism, energy production/conversion, and amino
acid transport/metabolism. Other 158 target genes (34%) comprise the accessory regulon, most of which were
related to cell wall structure/biogenesis, and virulence factor pathways. The remaining target genes (58%) were
unique to each strain, with largely unknown gene functions. The pan-regulon is applied to: 1) link strain-specific
phenotypes (growth imparied, siderophore production, antibiotic resistance) to underlying molecular mechanisms;
and 2) reveal that Fur has a disparate of target genes between the nine strains. Taken together, the Fur
transcriptional regulation varies in the same E. coli species, though the binding pattern is broadly conserved. We
thus, for the first time, provide experimental evidence of the Fur pan-regulon that shows surprisingly high
diversity in regulon genes amongst strains of the same species.

Keywords : Pan-genome, Pan-regulon, transcription factor binding sites, transcription regulation network

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S463 Synthetic Biosensor Accelerates Evolution by Rewiring Carbon Metabolism toward Specific
Metabolite

Sang Woo SEO

School of Chemical and Biological Engineering, Seoul National University, Seoul, Korea
Corresponding Author Email : [email protected]

Proper carbon flux distribution between cell growth and production of a target compound is important for
biochemical production because improper flux reallocation inhibits cell growth, thus adversely affecting
production yield. Here, using a synthetic biosensor to couple production of a specific metabolite with cell
growth, we spontaneously evolved cells under the selective condition toward the acquisition of genotypes that
optimally reallocated cellular resources. Using 3-hydroxypropionic acid (3-HP) production from glycerol in
Escherichia coli as a model system, we combined genome-seq, RNA-seq, and ChIP-exo analyses, and deter-
mined that mutations in the conserved regions of proteins involved in global transcriptional regulation altered
the expression of several genes associated with central carbon metabolism. These changes rewired central
carbon flux towards the 3-HP production pathway, increasing 3-HP yield and reducing acetate accumulation by
alleviating overflow metabolism. Our study provides a new perspective on adaptive laboratory evolution (ALE)
using synthetic biosensors, thereby supporting future efforts in metabolic pathway optimization.

Keywords : NGS, Biosensor, ALE

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S464 Sensitive One-pot Isothermal Detection of Pathogen-derived RNAs

Jeong Wook LEE

Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Korea
Corresponding Author Email : [email protected]

The recent outbreaks of Ebola, Zika, MERS, and SARS-CoV-2 (2019-nCoV) require fast, simple, and sensitive
onsite nucleic acid diagnostics that can be developed rapidly to prevent the spread of diseases. We have
developed a SENsitive Splint-based one-step isothermal RNA detection (SENSR) method for rapid and simple
onsite detection of pathogen RNAs with high sensitivity and specificity. SENSR consists of two simple
enzymatic reactions: a ligation reaction by SplintR ligase and subsequent transcription by T7 RNA polymerase.
The resulting transcript forms an RNA aptamer that induces fluorescence. Here, we demonstrate that SENSR is
an effective and highly sensitive method for the detection of the current epidemic pathogen, severe acute
respiratory syndrome-related coronavirus 2 (SARS-CoV-2). We also show that the platform can be extended to
the detection of five other pathogens. Overall, SENSR is a molecular diagnostic method that can be developed
rapidly for onsite uses requiring high sensitivity, specificity, and short assaying times.

Keywords : SASR-CoV-2, molecular diagnosis, isothermal, one-pot

45
2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S4-7]
Frontiers in Biomedical Studies

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S471 Biochemical Engineering in Animal Cell Technology

Takeshi OMASA
Department of Biotechnology Graduate School of Engineering, Osaka University, Osaka, Japan
Corresponding Author Email : [email protected]

Therapeutic antibodies are now the workhorse for the pharmaceutical industry in 21 century. Mammalian cell
lines are important host cells for the industrial production of therapeutic antibodies because of their capacity for
correct folding, assembly, and post-translational modification. In particular, Chinese hamster ovary (CHO) cells
are the most dependable host cells for the industrial production of therapeutic antibodies. In this presentation, I
introduced our recent approach for CHO genome instability, a bottle-neck process for antibody production, and
to prevent antibody aggregation in CHO cell cultivation. Moreover, I will introduce the integrated research and
developments in manufacturing technologies for therapeutic protein and cell and gene therapy at the MAB
organization.

Keywords : biochemical engineering, mammalian cell, biopharmaceuticals, Chinese hamster ovary cell
References
1. K. Kaneyoshi, et al., J. Biosci. Bioeng. 127: 107-113 (2019).
2. M. Onitsuka*, et al., J. Biosci. Bioeng., 127:752-757 (2019).
3. N. Yamano-Adachi*, et al., J. Biosci. Bioeng., 129:121-128 (2020).
4. T. Omasa, J. Biosci. Bioeng., 94:600-605 (2002). (Review).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S472 Epidural Electrotherapy for Brain Disorders

Sunggu YANG
Department of Nano-bioengineering, Incheon National University, Incheon, Korea
Corresponding Author Email : [email protected]

Penetrating electronics have been used for treating epilepsy, yet their therapeutic effects are debated largely due
to the lack of a large scale, real-time, safe recording/stimulation. Here, our proposed technology integrates
ultrathin epidural electronics into an electrocorticography (ECoG) array, therein simultaneously sampling brain
signals in a large area for diagnostic purposes and delivering electrical pulses for treatment. The system was
empirically tested to record the ictal-like activities of the thalamocortical network in vitro and in vivo using our
epidural electronics. Also, it is newly demonstrated that the electronics selectively diminish epileptiform
activities, but not normal signal transduction, in live animals. We propose that this technology heralds a new
generation of diagnostic and therapeutic brain-machine interfaces. Such an electronic system can be applicable
for several brain diseases such as tinnitus, Parkinson’s disease, Huntington’s disease, depression, and schizo-
phrenia.

Keywords : Electrotherapy, Brain disorders, Graphene, E-skin

References
1. SW. Park, et al., Small 14 (30) (2018).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S473 Development of in vitro Human Skeletal Muscle Models Using Microfabricated Devices

Kazunori SHIMIZU
Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Nagoya, Japan
Corresponding Author Email : [email protected]

In vivo animal experiments have been employed to understand disease mechanisms and check efficacy, toxicity
and irritancy of drugs, chemicals, and cosmetics. However, there is a growing need for alternatives to the animal
experiments because of the difference of physiological responses between animals and humans, and ethical
issues for the use of animals. One of the major alternatives is an in vitro cell culture. Although two-dimensional
(2D) cell culture has been employed conventionally, the cultured cells in flat layers on plastic surfaces does not
accurately model the in vivo state. In addition, it is difficult to evaluate tissue/organ-like functions including
barrier functions, adsorption functions and contractile properties by the conventional 2D culture technique.
Thus, better in vitro cell culture techniques to mimic and evaluate tissue/organ-level functions are needed.
Skeletal muscle is the most abundant tissue in our body and does not only function as an actuator but involves in
multiple physiological functions such as glucose homeostasis and body temperature maintenance, which are all
influenced by muscle diseases. Therefore, the functional deterioration of skeletal muscles directly causes a
reduced exercise capacity as well as metabolic abnormality, in turn worsening the quality of life significantly.
Especially, in super aging society, diseases causing muscle weakness including sarcopenia and disuse atrophy
are received much attention.
We have developed in vitro human skeletal muscle models (1-7). In this presentation, I will mainly talk about 1)
microdevices for contractile force measurement of tissue engineered skeletal muscle and their application for
phenotypic drug screening against skeletal muscle atrophy and 2) microdevices for contractile force measure-
ment of tissue engineered skeletal muscle innervated by iPSC derived motor neurons (7).

Keywords : In vitro models, Skeletal muscle cells, Motor neurons, Human iPSCs, Contractile force, Tissues/
Organs-on-a-Chip
References
1. Shimizu, K., Genma, R., Goto, Y., Nagasaka, S., Honda, H.: Bioengineering, 4(2):56 (2017).
2. Yamaoka, N., Shimizu, K., Imaizumi, Y., Ito, T., Okada, Y., Honda, H.: BioChip Journal, 13(2): 127-132
(2019).
3. Shimizu, K., Ohsumi, S., Kishida, T., Mazda, O., Honda, H.: Journal of Bioscience and Bioengineering,
129(5): 632-637 (2020).
4. Nagashima, T., Hadiwidjaja, S., Ohsumi, S., Murata, A., Hisada, T., Kato, R., Okada, Y., Honda, H.,
Shimizu, K.: Advanced Biosystems, 4: 2000121 (2020).
5. Ding, R., Horie, M., Nagasaka, S., Ohsumi, S., Shimizu, K., Honda, H., Nagamori, E., Fujita, H.,
Kawamoto, T.: Journal of Bioscience and Bioengineering, 130(1): 98-105 (2020).
6. Yoshioka, K., Ito, A., Arifuzzaman, Md., Yoshigai, T., Fan, F., Sato, K., Shimizu, K., Kawabe, Y.,
Kamihira, M.: Journal of Bioscience and Bioengineering, in press.
7. Yamamoto, K., Yamaoka, N., Imaizumi, Y., Nagashima, T., Furutani, T., Ito, T., Okada, Y., Honda, H.,
Shimizu, K.: bioRxiv, doi.org/10.1101/2021.01.07.424253.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S474 Dissolving Microneedle for Chronic Inflammatory Skin Diseases

Hyungil JUNG
Department of Biotechnology, Yonsei University, Seoul, Korea
Corresponding Author Email : [email protected]

Triamcinolone acetonide (TA) is one of the most widely prescribed drugs for relieving atopic dermatitis (AD).
Since its conventional injection method referred to as TA intralesional injection causes severe pain to patients,
applying dissolving microneedles (DMNs) on skin to deliver TA can be an alternative method for treating AD.
In this presentation, I introduce our recent approach to overcome TA’s poor dissolving nature which makes it
difficult to design and fabricate DMNs containing therapeutic dosage of TA. Moreover, I will introduce the
design and fabrication of candlelit shaped TA-loaded DMNs (Candlelit-DMN) which enabled TA evenly
distributed across target layer in human skin tissue, and effectively alleviated inflammation.

Keywords : Triamcinolone acetonide, Atopic dermatitis, Dissolving microneedles, Candlelit-DMN

References
1. M. Jang, et al., Adv. Healthcare Mater., 202001691 (2021).
2. M. King, et al., J. Clin. Aesthet. Dermatol., 13.1: E53 (2020).
3. M. J. Ho, et al., Pharmaceutics., 11.8:419 (2019).
4. Y.C. Kim, et al., Adv. Drug Delivery., 64:1547 (2012). (Review)
5. S.F. Lahiji, et al., Sci. Rep., 9:1 (2019).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S4-8]
Systems Biology for Industrial Applications

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S481 The Molecular Treasure Hunt – An ‘Omics-based Approach to Find New Bioactive Compounds

Tilmann WEBER
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kgs. Lyngby,
Denmark
Corresponding Author Email : [email protected]

Genome analyses of many microorganisms but also higher organisms indicate that the genetic potential to
synthesize specialized metabolites is far beyond the number of molecules observed in traditional screenings.
With the availability of cheap and easy-to-obtain whole genome sequences, in silico genome mining has become
an indispensable tool to complement the classical chemistry-centered approach to identify and characterize
novel secondary / specialized metabolites. Since the initial release in 2011, the open source genome mining
pipeline antiSMASH(1) (https://antismash.secondarymetabolites.org), which we develop in collaboration with
the group of M. Medema (U. Wageningen, Netherlands) and many international contributors, has become one of
the most widely used tools. We recently released version 6 of antiSMASH, including an improved user
interface, new detection modules, a new cluster comparison tool, and many internal optimizations. Specialist
and non-specialist users can easily analyze genomic sequences for the presence of secondary metabolite
biosynthetic gene clusters with antiSMASH.
To provide extensive analysis options of the data generated with antiSMASH, we have extended the framework
with several databases (2-4). The antiSMASH database, (https://antismash-db.secondarymetabolites.org/)(2),
contains 147,517 high quality BGC regions from 388 archaeal, 25,236 bacterial and 177 fungal “high-quality”
genomes.
These genome mining technologies build the foundation of further in silico studies towards a more compre-
hensive “Genome Analytics” platform, which we use to streamline our natural product discovery and charac-
terization efforts.
Albeit streptomycetes are studied for many decades as proficient producers of bioactive compounds, there are
still severe limitations concerning efficiency of mutagenesis protocols that often hamper systems metabolic
engineering and Synthetic Biology approaches. We have therefore developed an extensive CRISPR/Cas9-based
toolkit (5-7) for streptomycetes that now also includes tools that utilize multiplexing and DSB-free base editing
technology to highly effectively engineer actinomycetes.

Keywords : Genome mining, antibiotics, actinomycetes, Streptomyces, CRISPR

References
1. Blin, K., Shaw, S., Steinke, K., Villebro, R., Ziemert, N., Lee, S.Y., Medema, M.H. and Weber, T. (2019)
antiSMASH 5.0: updates to the secondary metabolite genome mining pipeline. Nucleic Acids Res., 47,
W81-W87.
2. Blin, K., Shaw, S., Kautsar, S.A., Medema, M.H. and Weber, T. (2021) The antiSMASH database version
3: increased taxonomic coverage and new query features for modular enzymes. Nucleic Acids Res., 49,
D639-D643.
3. Kautsar, S.A., Blin, K., Shaw, S., Weber, T. and Medema, M.H. (2021) BiG-FAM: the biosynthetic gene
cluster families database. Nucleic Acids Res., 49, D490-D497.
4. Kautsar, S.A., Blin, K., Shaw, S., Navarro-Munoz, J.C., Terlouw, B.R., van der Hooft, J.J.J., van Santen,
J.A., Tracanna, V., Suarez Duran, H.G., Pascal Andreu, V. et al. (2020) MIBiG 2.0: a repository for
biosynthetic gene clusters of known function. Nucleic Acids Res., 48, D454-D458.
5. Tong, Y., Whitford, C.M., Blin, K., Jorgensen, T.S., Weber, T. and Lee, S.Y. (2020) CRISPR-Cas9,
CRISPRi and CRISPR-BEST-mediated genetic manipulation in streptomycetes. Nat. Protoc., 15, 2470-
2502.
6. Tong, Y., Whitford, C.M., Robertsen, H.L., Blin, K., Jorgensen, T.S., Klitgaard, A.K., Gren, T., Jiang, X.,
Weber, T. and Lee, S.Y. (2019) Highly efficient DSB-free base editing for streptomycetes with CRISPR-
BEST. Proc. Natl. Acad. Sci. U. S. A., 116, 20366-20375. 7. Tong, Y., Charusanti, P., Zhang, L., Weber, T.
and Lee, S.Y. (2015) CR
7. Tong, Y., Charusanti, P., Zhang, L., Weber, T. and Lee, S.Y. (2015) CRISPR-Cas9 based engineering of
actinomycetal genomes. ACS Synth. Biol., 4, 1020-1029.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S482 Systems Biological Model-guided Design and Engineering of Personalized Probiotics with Host
and Microbiome Interactions

Dong-Yup LEE
School of Chemical Engineering, Sungkyunkwan University, Suwon, Korea
Corresponding Author Email : [email protected]

Probiotics are live beneficial microorganisms that can be consumed in the form of dairy and food products as
well as dietary supplements to promote a healthy balance of gut bacteria in humans. Now, one of the main
scientific and industrial challenges is to identify and select promising strains and formulate multi-strain
probiotic blend with consistent efficacy which is highly dependent on individual dietary regimes, gut
environments, and health conditions [1]. Interestingly, the limitations of current in vivo and in vitro methods for
testing probiotic strains can be overcome by in silico model guided systems biology approach since genome
scale metabolic models (GEMs) can be used to describe and predict their cellular behaviours, metabolic states
and community interactions under various gut environments. In this talk, I will summarize currently available
GEMs of microbial strains with probiotic potentials, and present a knowledge-based platform to evaluate their
capabilities on the basis of probiotic criteria in terms of metabolic characteristics, stability, safety, colonization,
postbiotics, interaction with the gut microbiome and variation across diets. Its applicability is demonstrated by a
case study where genome-wide metabolic landscapes in several lactic acid bacteria (LAB) are systematically
evaluated to elucidate diet-induced and strain specific probiotic features. In addition, I will show how their
metabolic interactions with the host and gut microbiome can be investigated to propose new strategies for
personalized probiotics design, thereby ameliorating lifestyle diseases.

Keywords : Systems biology, smart probiotics, lactic acid bacteria (LAB), microbiome, genome-scale metabolic
model
References
1. Choi, Y. M., Y. Q. Lee, H.-S. Song and D.-Y. Lee. 2020. Genome scale metabolic models and analysis for
evaluating probiotic potentials. Biochem. Soc. Trans., 48(4): 1309-1321.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S483 Systems Biology Applications in Precision Medicine: Multi-omics Approaches

Sunjae LEE
School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, Korea
Corresponding Author Email : [email protected]

“data-driven” life sciences. Recent advance in high throughput technology has enabled us in-depth molecular
profiling of individuals with reasonable economic costs and provided the integrated understanding of clinical
trials and disease progressions. Systems biology tools have accelerated the integration of multi-omics data and
helped the identifications of druggable targets and biomarkers of chronic diseases.

For example, molecular signatures of hundred individuals has been traced over a year (Swedish wellness study)
and found significant signatures of pre-disease states by systems biology approaches (Abdella Tebani et al.,
2020, Nature Communications). Clinical interventions of fatty liver diseases by isocaloric low carbohydrate
diets was explored with multi-omics data and elucidated underlying molecular mechanisms that boosted the fat
burning during the interventions (Adil Mardinoglu et al., 2018, Cell Metabolism). Furthermore, systems biology
identified the biomarkers of insulin resistance, plasma mannose, which was found more powerful than blood
glucose, and druggable targets of fatty liver diseases, pyruvate kinase L/R (Sunjae Lee et al., 2017, Molecular
Systems Biology; Sunjae Lee et al., 2016, Cell Metabolism).

In this talk, the speaker will address the current systems biology applications in precision medicine, especially
multi-omics analysis of chronic diseases. Especially recent efforts of understanding human gut microbiome by
systems biology approaches, such as pan-disease metagenomic association studies, will be presented.

Keywords : systems biology, precision medicine, human metabolism, human microbiome

References
1. Abdellah Tebani et al., “Integration of molecular profiles in a longitudinal wellness profiling cohort”
(2020), Nature Communications.
2. Adil Mardinoglu et al., “An integrated understanding of the rapid metabolic benefits of a carbohydrate-
restricted diet on hepatic steatosis in humans” (2018), Cell Metabolism.
3. Sunjae Lee et al., “Network analyses identify liver‐specific targets for treating liver diseases” (2017),
Molecular Systems Biology.
4. Sunjae Lee et al., “Integrated network analysis reveals an association between plasma mannose levels and
insulin resistance” (2016), Cell Metabolism.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S484 Retrobiosynthesis for Metabolic Engineering

Hyun Uk KIM
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology
(KAIST), Daejeon, Korea
Corresponding Author Email : [email protected]

Retrobiosynthesis helps systematically design novel biosynthetic pathways for the production of chemicals and
materials through metabolic engineering. Retrobiosynthesis is typically implemented by using a set of
biochemical reaction rules that describe chemical transformation patterns between substrates and products at an
atomic level. Despite its great potential, experimental validation of the retrobiosynthetic predictions has been
conducted only for a few chemicals. Here, application of retrobiosynthesis for the production of multiple short-
chain primary amines (SCPAs) using Escherichia coli will be presented [1]. SCPAs have a wide range of
industrial applications, for example, as a precursor of pharmaceuticals, agrochemicals, and solvents. Also, a
deep learning model named DeepRFC will be introduced that examines the feasibility of a large number of
retrobiosynthesis-derived enzymatic reactions [2]. DeepRFC will help reduce the number of the initially
predicted reactions for experimental validation. These studies will allow more active application of retrobio-
synthesis in metabolic engineering.

Keywords : Metabolic engineering, Systems biology, Retrobiosynthesis, Short-chain primary amines, DeepRFC
References
1. Kim et al. Microbial production of multiple short-chain primary amines via retrobiosynthesis. Nature
Communications 12, 173 (2021).
2. Kim et al. A deep learning approach to evaluate the feasibility of enzymatic reactions generated by
retrobiosynthesis. Biotechnology Journal. In press.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S4-9]
New Application of Biotechnology
in Food Industry

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S491 Engineering Yeast Strains for Producing Food Ingredients and Value-added Chemicals

Eun Joong OH
Department of Food Science, Purdue University, West Lafayette, USA
Corresponding Author Email : [email protected]

Microbes have recently been engineered to be cellular factories with metabolic pathways designed to convert
carbon sources, such as sugars, into value-added chemicals and food materials. However, not all fermentation
processes are currently applicable for industrial-scale production. This is due to several challenges, such as high
feedstock costs and low productivity. Metabolic engineering is an applied science that employs recombinant
DNA technology to improve cellular activity. Metabolic engineering has been advanced as an effective tool with
which to improve industrial fermentation processes. My talk will focus on fermentation science and the
biotechnological production of fuels and value-added chemicals using engineered yeast strains. I propose
metabolic engineering and fermentation research using rational and combinatorial tools to develop and optimize
engineered strains capable of rapid and efficient fermentation of abundant sugars in non-food plant biomass,
including cellobiose and xylose. I will also highlight multiplex CRISPR/Cas9 gene editing coupled with high-
throughput screening and deep sequencing in the field of genome engineering, leading to a better understanding
of genotype-phenotype correlations.

Keywords : Yeast, metabolic engineering, CRISPR/Cas9

References
1. Oh, E. J., Liu, R., Liang, L., Freed, E. F., Eckert, C. A., and Gill, R. T. 2020. Multiplex evolution of
antibody fragments utilizing a yeast surface display platform. ACS Synthetic Biology 9(8), 2197-2202.
2. Oh, E. J., Skerker, J. M., Kim, S. R., Wei, N., Turner, T. L., Maurer, M., Arkin, A. P., and Jin, Y. S. 2016.
Gene amplification on demand accelerates cellobiose utilization in engineered Saccharomyces cerevisiae.
Applied and Environmental Microbiology 82, 3631-3639.
3. Oh, E. J., Ha, S. J., Kim, S. R., Lee, W. H., Galazka, J. M., Cate, J. H. D., and Jin, Y. S. 2013. Enhanced
xylitol production through simultaneous co-utilization of cellobiose and xylose by engineered Saccha-
romyces cerevisiae. Metabolic Engineering 15, 226-234.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S492 Biosynthesis of Functional Food Materials by Amylosucrase

Dong-Ho SEO
Department of Food Science and Technology, College of Agriculture and Life Sciences, Jeonbuk National
University, Jeonju, Korea
Corresponding Author Email : [email protected]

Amylosucrase (ASase; EC 2.4.1.4) is an enzyme that has great potential in the biotechnology and food
industries, due to its multifunctional enzyme activities. It can synthesize α-1,4-glucans, like amylose, from
sucrose as a sole substrate, but importantly, it can also utilize various other molecules as acceptors. It efficiently
synthesizes modified starch with increased ratios of slow digestive starch (SDS) and resistant starch (RS), and
glucosylated functional compounds with increased water solubility and stability. First, novel modified chestnut
starch was synthesized by ASase from Deinococcus geothermalis (DGAS). DGAS-modification of chestnut
starch increased non-digestible resistant starch and it ameliorated diet-induced obesity via GPR43-mediated
suppression of insulin signaling thereby presumably reducing fat accumulation in white adipose tissue. Second.
α-flavone glycosides have beneficial properties for applications in the pharmaceutical, cosmetic, and food
industries. However, their chemical syntheses are often limited by a low efficiency or scarcity of substrates. α-
flavone glucosides were enzymatically synthesized by DGAS using sucrose and various flavones as a donor for
glucosyl units and acceptors, respectively. Finally, the DGAS expressed in Corynebacterium glutamicum
(cDGAS) and purified via Ni-NTA affinity chromatography were compared to those of DGAS expressed in
Escherichia coli (eDGAS). The pH profile of cDGAS was similar to that of eDGAS, whereas the temperature
profile of cDGAS was lower than that of eDGAS. The melting temperature of both enzymes did not differ
significantly. Interestingly, polymerization activity was slightly lower in cDGAS than in eDGAS, whereas
luteolin (an acceptor molecule) transglucosylation activity was greater in cDGAS than in eDGAS. Furthermore,
The transglycosylation activity of cDgAS was optimized for luteolin glycoside synthesis using purified enzymes
(PEs), whole cells (WCs) suspended in buffer (WCB), and WCs in ethanol (WCE). The reaction parameters for
PE, WCE, WCB, and IC transglycosylation were optimized using response surface methodology, and ICs
required lower concentrations of substrates while synthesizing higher concentrations of reaction products in
addition to producing the novel compound luteolin maltotrioside after several cycles of reuse.

Keywords : Amylosucrase, Transglycosylation, Deinococcus geothermalis, Corynebacterium glutamicum

References
1. D. H. Seo, S. H. Yoo, S. J. Choi, Y. R. Kim and C. S. Park, Food Sci. Biotechnol. 29(1):1-16 (2020).
2. E. S. Lee, B. H. Lee, D. U. Shin, M. Y. Lim, W. H. Chung, C. S. Food Hydrocolloids. 75:22-23 (2018)Park,
M. Y. Baik, Y. D. Nam, D. H. Seo.
3. S. W. Jang, C. H. Cho, Y. S. Jung, C. Rha, T. G. Nam, D. O. Kim, Y. G. Lee, N. I. Baek, C. S. Park, B. H.
Lee, S. Y. Lee, H. S. Shin, D. H. Seo, Plos One. 13(11): e0207466 (2018).
4. Y. W. Chin, S. W. Jang, H. S. Shin, T. W. Kim, S. K. Kim, C. S. Park, D. H. Seo, Enzyme Microb. Technol.
135:109505 (2020).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S493 Future Muscle Food : Status and Prospects of Cultured Meat

Cheorun JO
Department of Agricultural Biotechnology, Seoul National University, Seoul, Korea
Corresponding Author Email : [email protected]

Cellular agriculture is an emerging research field of agribiotechnology. Stem cell and tissue engineering makes
it possible to produce agricultural products and by-products without a sacrifice of animals. One of the
representative technologies in cellular agriculture, cultured muscle tissue-based protein products, known as
cultured meat, are produced through in vitro myogenesis involving muscle stem cell culture and differentiation,
and mature muscle cell processing for flavor and texture. The idea of cultured meat has been developed to
resolve some problems related to industrial livestock farming by lessening undesirable results from traditional
animal farming. In this presentation, the background of development, status and prospects of cultured meat will
be discussed at international and national levels.

Keywords : Cultured meat, muscle food, cellular agriculture, myogenesis

References
1. Mark J. Post, Shulamit Levenberg, David L. Kaplan, Nicholas Genovese, Jianan Fu, Christopher J. Bryant,
Nicole Negowetti, Karin Verzijden, and Panagiota Moutsatsou, Scientific, sustainability and regulatory
challenges of cultured meat (2020) Nature Food, 1, 403-415.
2. Kwang-Hwan Choi, Ji Won Yoon, Minsu Kim, Hyun Jung Lee, Jinsol Jeong, Minkyung Ryu, Cheorun Jo,
Chang-Kyu Lee, Muscle stem cell isolation and in vitro culture for meat production: A methodological
review (2021) Comprehensive Review in Food Science and Food Safety, 20, 429-457.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S494 Potential of Enhancing Cellulose Microfibrillation or Nanocrystallization to Use as a Functional

Biomaterial

Young Hoon JUNG

School of Food Science & Biotechnology, Kyungpook National University, Daegu, Korea
Corresponding Author Email : [email protected]

Cellulose is one of the most abundant biopolymers in the world. Nano-sized cellulose, which can be obtained
through extraction of either fibrils by physical treatment or crystals by chemical treatment from cellulose, has
much wide surface area in comparison with cellulose. Thereby, the nanocellulose can give special properties to
the final materials when using as an additive. In this talk, we prepared microfibrillated cellulose and nanocrystal
cellulose from a red algal biomass, Gelidium amansii. Through consecutive steps such as removal of other
compounds except cellulose and physical or chemical pretreatment, each form of nanocellulose was obtained.
Then, their surface characteristics including size dimensions, morphologies, degree of fibrillation, etc. were
widely investigated. Also, the effect of those nanocellulose as an anti-inflammatory material on human
keratinocytes and mice skin was evaluated. In conclusion, both microfibrillated cellulose and nanocrystal
cellulose derived from Gelidium amansii migh be promising cosmeceutical agents.

Keywords : Nanocellulose, Microfibrillated cellulose, Nanocrystal cellulose, Anti-inflammation, Cosmeceutical,

Gelidium amansii

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S495 Advanced Electrochemical Sensor Modified with Affinity Peptides for Sensitive Norovirus
Detection

Jong Pil PARK

Department of Food Science and Technology, Chung -Ang University, Anseong, Korea
Corresponding Author Email : [email protected]

Electrochemical-based biosensors have been widely applied in many fields because they have high sensitivity,
are easy to control, and allow for label-free detection and miniaturization in a point-of-care testing. However,
target-specific probes such as DNA, antibody, and protein on the working electrode surface is needed to enable
specific detection of targets in electrochemical analysis. Antibodies serve as important capture reagents in
several fields such as the clinical or pharmaceutical analysis, diagnosis, and treatment of various diseases.
However, they have limitations such as being relatively large in size and having cross-reactivity to be occurred
in immunoassay tests. In contrast, affinity peptides have a small size and simple structure and are relatively
stable in a variety of environmental conditions. The aim of this talk is to show unique affinity peptides which
would recognize norovirus with high specificity and selectivity and to explore the binding interactions using
several analytical methods. To improve the abilities of affinity peptides, we used a rational design approach with
in silico modeling and tried to synthesize advanced affinity peptides for an electrochemical sensor.

Keywords : affinity peptide, binding affinity, modification, engineering, norovirus detectioin

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

Special Program

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S5-1]
바이오 소재분야 부상기술 연구동향
분석사업 성과발표회

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S511 바이오 의약 소재

최종훈
중앙대학교 융합공학부 바이오메디컬공학전공
Corresponding Author Email : [email protected]

본 발표에서는 바이오 의약 분야 3 대 부상기술에 대해 논하고자 함. 나노바이오 기반 진단기술에서는

최근 부상하고 있는 생체 내 자연적으로 분비되는 세포외소포체 (엑소좀)를 이용한 질병의 조기진단
기술개발에 대해 현재의 상황과 미래를 예측하고자 함. 두 번째로 차세대 백신 기술로서 대두되고 있
는 mRNA 백신 기술의 현재 동향에 대해 논하고자 함. 마지막으로 성공적으로 유해 박테리아를 제거
하고 바이오필름의 형성을 예방하기 위해서 균주의 독성과 항생물질에 대해 내성이 생기는 문제점을
해결하고자 하는 항균 나노 신소재들에 대해 그 현재와 미래를 다뤄보고자 함.

Keywords : 세포외소포체 진단, mRNA 백신, 항균 나노 신소재

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S512 바이오 생리활성 분야 국내외 유망기술 및 산업 동향 분석

서승오
가톨릭대학교 식품영양학과
Corresponding Author Email : [email protected]

글로벌 팬더믹 시대에 인류의 건강 증진과 지속가능한 삶을 구현하기 위한 사회적 요구와 산업적 수
요가 증가함에 따라 감염병에 효과적으로 대처하는 다양한 유망 바이오 생리활성 소재 및 기반 기술
이 급격하게 성장하고 있다. 이러한 포스트 코로나 시대의 바이오산업 패러다임 변화에 대응하기 위
해 인류의 건강과 밀접한 관련이 있는 바이오 생리활성 소재 분야의 국내외 유망 기술 및 산업 동향
에 대해 분석하였다. 특히, 면역력 증강 등과 같은 건강기능성을 바탕으로 감염병에 대응할 수 있는
다양한 바이오 생리활성 소재를 생물공학적으로 제조하는 기술이 아래와 같이 3 대 부상기술로 선정
되었다.
1. 합성비타민 대체 바이오비타민 소재 제조기술
2. 미세조류 유래 차세대 생리활성 소재 제조기술
3. 글로벌 팬더믹 대응 미래 바이오 웰빙/메디칼 소재 제조기술
각 분야의 전문가들을 대상으로 한 설문조사와 더불어 국내외 최근 연구동향 및 산업동향 분석을 통
해 해당 부상 기술의 개념, 기술의 중요성, 산업적 필요성, 국내외 기술수준 및 격차, 정부지원 방안,
해외협력 방안 등을 도출하였다.

Keywords : 바이오소재 부상기술, 바이오 생리활성 소재, 바이오비타민, 미세조류, 웰빙/메디칼 소재

References
1. 바이오소재 분야 부상기술 연구동향 분석 연구 최종보고서, 한국생물공학회, 2021.03.15

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2021 KSBB Spring Meeting and International Symposium

S513 바이오 화학

김희택
충남대학교 농업생명과학대학 식품공학과
Corresponding Author Email : [email protected]

최근, 전 세계적으로 플라스틱 폐기물과 지구 온난화가 주요한 사회문제로 대두되고 있습니다. 이와

같은 문제를 해결하기 위해 생분해성 플라스틱 개발, 플라스틱 업사이클링 개발등 다양한 접근법들이
제시되고 있지만, 제한적인 범위의 플라스틱 생산 및 이산화탄소 배출 문제 등 여러 현안들이
남아있습니다. 이와 같은 문제를 근원적으로 해결하기 위한 부상기술에 대한 연구가 최근 활발히
이루어 지고 있습니다. 본 발표에서는 플라스틱 문제에 대한 현 상황을 공유하고 부상기술로서 Carbon
close loop 에 입각하여 도출된 바이오매스 기반 차세대 플라스틱 소재, 플라스틱 기능향상 소재 및
플라스틱의 생물학적 업사이클링 기술에 대하여 소개하도록 하겠습니다.

Keywords : 바이오매스, 플라스틱 소재, 기능향상 소재, 생물학적 업사이클링 기술

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2021 KSBB Spring Meeting and International Symposium

S514 바이오소재 분야 부상기술 연구동향 분석 – 바이오기반기술

연영주 1, 이대희 2, 안정오 3

1
강릉원주대학교 생명화학공학과, 2 한국생명공학연구원 합성생물학전문연구단,
3
한국생명공학연구원 바이오상용화지원센터
Corresponding Author Email : [email protected]

본 발표에서는 ‘바이오기반기술’ 분야의 3 대 부상기술에 대해 소개한다. 첫 번째 기술은 ‘AI 기반

스마트 바이오공정 개발 기술’로, 바이오소재 생산 관련 다양한 바이오공정으로부터 빅데이터 수집 및
플랫폼 모델 구축 빅데이터를 수집하고, 이를 활용하여 바이오공정별 과업을 스스로 판단해 실행할 수
있는 스마트 머신 개발과, 사물인터넷을 통한 스마트 머신 간의 상호 소통 체계 구축으로 최적의
바이오의약품 및 바이오소재 제품 생산을 위한 스마트 바이오팩토리를 구현하는 기술이다. 두 번째
기술은 ‘바이오파운드리 기반 산업용 고효율 미생물 초고속 개발 기술’로, 합성생물학 기술에 컴퓨터
기반 DNA 설계, 자동화, 인공지능을 융합하여 바이오산업의 오랜 난제인 속도, 규모, 효율을 혁신할 수
있는 바이오파운드리 시스템을 구축하고, 이를 산업용 고효율 미생물을 초고속으로 개발하고 상용화에
도입할 수 있는 기술 개발을 소개한다. 세 번째는 ‘차세대 분자설계 기술 기반 스마트 바이오촉매 개발
기술’로, 기존 분자모델링 및 단백질공학적 효소개량 기술에 빅데이터 및 인공지능 기술을 도입하여
기존 기술 대비 설계 정확도 개선, 개량 시간 단축, 투자비용 절감 등의 효과를 제공할 수 있는
분자설계 기술을 개발하고, 이를 통해 산업적 응용에서 요구되는 주요 성능지표들이 동시에 향상된
스마트 바이오 촉매를 개발하는 기술에 대해 소개한다. 상기 3 대 부상기술의 연구동향 및 중요성에
대해 소개할 것이며, 기술적 측면 뿐만 아니라 경제/사회적 및 정부/정책적 영향에 대해서도 논의될
것이다.

Keywords : 부상기술, 바이오기반기술, 인공지능, 빅데이터, 스마트 바이오공정, 바이오파운드리, 차세대

분자설계, 스마트 바이오촉매

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S5-2]
신진연구자를 위한 포럼

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S521 Peptide-Nanoparticle Conjugates for Immune Checkpoint Blockade

Woo-jin JEONG
Department of Biological Engineering, Inha University, Incheon, Korea
Corresponding Author Email : [email protected]

The majority of anti PD-1/PD-L1 agents currently approved or under development are based on full-size
monoclonal antibodies. Despite their demonstrated efficacy, the widespread use of the antibody drugs has been
hindered due to their high cost and complexity in manufacturing and low thermodynamic stability. Herein, we
introduce a peptide-dendrimer conjugate (PDC) approach that stabilizes the β-hairpin structure of peptides via
intermolecular forces and the excluded volume effect as well as exploits the multivalent binding effect. Because
of the synergistic advantages, the PDCs based on a β-hairpin peptide isolated from an engineered PD-1 protein
showed significantly higher affinity (avidity) to PD-L1, as compared to free peptides (by up to 5 orders of
magnitude). The enhanced binding kinetics with high selectivity was translated into an improved immune
checkpoint inhibitory effect in vitro, at a level comparable to (if not better than) that of a full-size monoclonal
antibody. The results demonstrate the potential of the PDC system as a novel class of inhibitors targeting β-
strand-rich protein surfaces, such as PD-1 and PD-L1, displaying its potential as a new cancer immunotherapy
platform.

Keywords : Peptide-nanoparticle conjugates, Cancer immunotherapy, Immune Checkpoint Blockade, Multivalent

binding effect, β-hairpin stabilization

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S522 Tailoring Biomaterials for Cancer Immunotherapy

Kye Il JOO
Department of Chemical Engineering and Materials Science, Ewha Womans University, Seoul, Korea
Corresponding Author Email : [email protected]

Despite the great promise of immune checkpoint blockade (ICB) therapy for cancer treatment, the currently
available options for ICB treatment pose major clinical challenges, including the risk of severe systemic auto-
immune responses. Here, we developed a novel localized delivery platform, immuno-bioglue (imuGlue), which
is inspired by the intrinsic underwater adhesion properties of marine mussels and can allow the optimal retention
of anti-PD-L1 drugs at tumor sites and the on-demand release of drugs in response to the tumor microenvi-
ronment. Using a triple-negative breast cancer and melanoma models, we found that imuGlue could signify-
cantly enhance anti-tumor efficacy by eliciting a robust T cell-mediated immune response while reducing
systemic toxicity by preventing the rapid diffusion of anti-PD-L1 drugs into the systemic circulation and other
tissues. It was also demonstrated that imuGlue could be successfully utilized for combination therapy with other
immunomodulatory drugs to enhance the anti-tumor efficacy of ICB-based immunotherapy, demonstrating its
versatility as a new treatment option for cancer immunotherapy.

Keywords : Biomaterial Design, Cancer Immunotherapy, Nano Immuno-Engineering

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S523 Nanotechnology Applied in Prevention, Diagnosis and Treatment of Infectious Diseases and
Cancer

Hyun-Ouk KIM1,2
1
Department of Biotechnology and Bioengineering, Kangwon National University, Chuncheon, Korea
2
Biohealth-machinery Convergence Engineering, Kangwon National University, Chuncheon, Korea
Corresponding Author Email : [email protected]

The worst pandemic in history was the Black Death, which killed a third of the European population in the
Middle Ages. Following the Spanish flu in 1918, the Asian flu in 1957, the Hong Kong flu in 1968, and
influenza A (H1N1), also called the swine flu in 2009, the World Health Organization (WHO) declared a
pandemic over COVID-19 after 11 years.
Despite the development of diagnosis, treatment, and vaccines for infectious diseases around the world, overseas
movement of people and animals is becoming easier, and the occurrence of emerging, newly emerging, and
exotic malignant contagious infectious diseases is increasing due to urban development, environmental
destruction, and so on.
Artificial nanostructures have not only physical properties with high surface area, but also various physic-
chemical properties (conductivity, catalytic activity, enzyme activity, optical properties, etc.), and can support
vaccines and therapeutic agents. Since the formation of various nanostructures can be used for prevention,
diagnosis and treatment, the convergence of nano and biotechnology is very important.
Therefore, there is a need to solve the problem of prevention, early diagnosis, and treatment of emerging, newly
emerging, exotic malignant contagious infectious diseases with nano and biotechnology convergence technology.

Keywords : Cancer, Diagnosis, Infectious Diseases, Nanoparticle, Prevention, Treatment

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S524 Cationic Polymer Development for mRNA Delivery and their Application for CRISPR-Cas
Technology

HyunJin KIM
Department of Biological Engineering, College of Engineering, Inha University, Incheon, Korea
Corresponding Author Email : [email protected]

Efficient nonviral vector gradually become an essential requisite for successful gene editing therapy via
CRISPR-Cas technology, which Cas9 protein edits guide RNA (gRNA)-complementary target chromosome
DNA. The nonviral vector in the local administration can accomplish gene editing in the area of interest without
inducing undesirable gene editing in the off-region. We have developed amphiphilic polyaspartamide deriva-
tives (PAsp(DET/CHE)) to induce transient and high Cas9 protein expression from messenger RNA (mRNA).
Diethylenetriamine (DET) moiety provides high endosomal escape ability and associates with negatively
charged mRNA. 2-Cyclohexylethyl (CHE) moiety optimizes polymer hydrophobicity between mRNA
nanoparticle stability in the extracellular condition and mRNA release in the cytoplasm. CHE moiety was
selected from the highest luciferase mRNA expression in cultured cells among ~20 aliphatic groups between 5
and 10 carbon atoms. PAsp(DET/CHE) was synthesized via two-step process. The precursor polymer poly(β-
benzyl-L-aspartate) (PBLA) were synthesized with ring opening polymerization between n-butylamine and β-
benzyl-L-aspartate N-carboxy-anhydride (BLA-NCA) to have degree of polymerization (DP) of 26.
PAsp(DET/CHE) was synthesized by co-aminolysis of DET and CHE into PBLA to contain 15 DET and 11
CHE moieties. Interestingly, the mRNA translation efficacy was not affected by DP of PAsp(DET/CHE) from
26 to 121 but by the amounts of CHE moieties in the single polymer. The PAsp(DET/CHE) with less than 8
CHE introduction amounts did not increase mRNA translation efficacy at all but dramatically enhanced the
translation efficacy with more than 11 CHE introduction amounts.
We analyzed in vitro cellular uptakes, endosomal escape ability, and release kinetics of mRNA in cytoplasm to
clarify the mechanism of high mRNA translation of PAsp(DET/CHE). Gene editing was evaluated in Ai9
mouse, which red fluorescence occurs in the results of Cas9/gRNA based STOP cassette cleavages. The nano-
particles were prepared with mixing between PAsp(DET/CHE) and Cas9 mRNA (or gRNA). The nanoparticles
(1.5 ug Cas9 mRNA and 1.5 ug sgRNA) were locally injected into ventricular area of the Ai9 mouse. At 2 day
post-administration, the brain was dissected and observed by using confocal laser scanning microscope (CLSM).
Gene editing occurred in the ependymal layer and choroid plexus of almost all brain area, which indicated high
delivery efficiency of PAsp(DET/CHE) into the brain. PAsp(DET/CHE) has high gene editing efficacy in the
local delivery into brain, nose, and muscle. These indicate high potential of the hydrophobicity optimization of
polyaspartamide derivatives and demonstrates the strong potential of PAsp(DET/CHE) for in vitro and in vivo
Cas9 mRNA/gRNA delivery.

Keywords : mRNA delivery, cationic polymer, gene editing, amphiphile

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S525 Ribocomputing: Leveraging RNA for Computation in the Cell

Jongmin KIM
Department of Life Sciences, Pohang University of Science and Technology, Pohang, Korea
Corresponding Author Email : [email protected]

Synthetic biology aims to develop engineering-driven approaches to the programming of cellular functions that
could yield transformative technologies. Synthetic gene circuits that combine DNA, protein, and RNA com-
ponents have demonstrated a range of functions such as bistability, oscillation, feedback, and logic capabilities.
Despite many advances, technical challenges remain for scaling up the complexity of these networks due to the
limited number of designable, orthogonal, high-performance parts, the empirical and often tedious composition
rules, and substantial resource requirements for encoding and operation. Here, we report a strategy for
constructing RNA-only nanodevices to evaluate complex logic in living cells. Such ‘ribocomputing’ systems are
composed of de novo designed parts and operate via predictable and designable base-pairing rules, allowing for
effective in silico design of computing devices with prescribed configurations and functions in complex cellular
environments. We demonstrate that these ribocomputing devices in Escherichia coli can evaluate two-input
logic expressions with dynamic range up to 900-fold and scale them to four-input AND, six-input OR, and a
complex 12-input logic expression [1]. We further demonstrate that ribocomputing design strategy can be used
to develop a large library of high-performance translational repressors and 4-input NAND logic gates [2].
Successful operation of ribocomputing devices based on programmable RNA interactions suggests that systems
employing the same design principles could be implemented in other host organisms or in extracellular settings.

Keywords : RNA synthetic biology, logic circuits, molecular computation

References
1. Alexander A. Green, Jongmin Kim, et al., Complex cellular logic computation using ribocomputing devices
(2017), Nature, 548(7665), 117-121.
2. Jongmin Kim, et al., De novo-designed translation-repressing riboregulators for multi-input cellular logic
(2019), Nature Chemical Biology, 15(12), 1173-1182.

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2021 KSBB Spring Meeting and International Symposium

S526 Synthetic Evolutionary Bioengineering: Cooperation of Nature’s Power and Human Design

Sungho JANG1,2
1
Division of Bioengineering, Incheon National University, Incheon, Korea,
2
Department of Bioengineering and Nano-Bioengineering, Incheon National University, Incheon, Korea
Corresponding Author Email : [email protected]

Biological systems have the potential to provide solutions for urgent challenges that we are facing from energy,
environment, and human health. Engineering biological systems, however, frequently involves screening of
extremely large solution space in the hope of finding useful phenotypes such as proteins with new functions or
metabolic pathways for chemical production. In this talk, I am going to introduce examples showing how human
design can cooperate with nature’s power to provide valuable solutions, focusing on the metabolic engineering
of microorganisms. In particular, the development and applications of synthetic regulatory RNAs will be
discussed which were utilized as molecular tools to guide the evolution of metabolite-producing recombinant
bacteria. Additionally, the rationally-designed molecular program for rapid diagnosis of pathogens called
SENSR will be discussed.

Keywords : Synthetic Biology, Artificial Evolution, Metabolic Engineering, Molecular Diagnosis

References
1. S. Jang, S. Jang, Y. Xiu, T. J. Kang, S.-H. Lee, M.A.G. Koffas, G. Y. Jung, ACS Synth. Biol. 6(11), 2077-
2085 (2017).
2. S. Jang, S. Jang, D.-K. Im, T. J. Kang, M.-K. Oh, G. Y. Jung, ASC Synth. Biol. 8(6), 1729-1283 (2019).
3. S. Jang, S. Jang, J. Yang, S. W. Seo, G. Y. Jung, Curr. Opin. Biotechnol. 53, 1-11 (2018).
4. H. G. Lim, S. Jang, S. Jang, S. W. Seo, G. Y. Jung, Curr. Opin. Biotechnol. 54, 18-25 (2018).
5. C. H. Woo, S. Jang, G. Shin, G. Y. Jung, J. W. Lee, Nat. Biomed. Eng. 4(12), 1168-1179 (2020).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S527 Expanding the Limits of the Second Genetic Code with Ribozymes

Joongoo LEE
Department of Chemical Engineering, POSTECH, Pohang, Korea
Corresponding Author Email : [email protected]

From a chemical engineer’s perspective, biology is technology. As an example, biology produces materials that
no other human-made technology and chemistry can, and operates numerous complex chemical/enzymatic
reactions across a wide variety of lengths of scales from the single-molecule to the macro-scale level. Thus,
scientists have engineered living organisms to harness the advanced technologies that are embedded in biology
and manufacture next-generation commodities on demand. However, engineering living organisms to produce
high-value molecules remains costly and slow, and new tools are needed to understand and expand the powerful
capabilities of biotechnology. In this presentation, I will discuss my strategy to address this issue and broaden
the scope of biological material-catalyzed transformations.

In the first portion of my talk, I will describe our recent efforts to expand the genetic code using ribozymes for
the synthesis of bio-based products that extend beyond natural limits. In the second part, I will explain my
investigation into engineering the translational apparatus that has been evolutionarily optimized to accept natural
amino acid building blocks. Overall, my work aims to deepen a fundamental understanding of the origin of life
and enable new classes of functional materials such as precision polymers, therapeutics, and biosensors.

Keywords : ribosome, ribozyme, unnatural amino acids, non-canonical chemical substrates, cell-free protein
expression, protein translation, DNA, RNA, polymers, non-amide backbone

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S528 Repetitive Sequence Analysis of Fibrous Proteins Using Bioinformatics and the Roles of KISTEP
for Researchers

Dooyup JUNG
Center for Growth Engine R&D Coordination, Korea Institute of S&T Evaluation and Planning, Eumseong,
Korea
Corresponding Author Email : [email protected]

This talk will be organized as two parts. The first part is the amino acid sequence analysis of fibrous proteins
harboring repeating units using bioinformatics. Because there are close relationships among sequences,
molecular structures, and mechanical properties, appropriate computational methods are necessary to efficiently
analyze, find, store, and utilize fibrous protein sequences. However, pre-established bioinformatical tools do not
suitably contribute to fibrous proteins due to their sequence repetitiveness. Therefore, in this talk, the
development of novel bioinformatical platform for fibrous protein sequences will be introduced from spider silk
sequence analysis with spider evolution and fiber property to fibrous protein fingerprint database construction.
The second part is about the roles of KISTEP to support R&D policy. Especially, the process and method of
government R&D budget allocation and coordination in Korea will be briefly explained, mainly dealing with
materials field as an example. In addition, the information produced or edited by KISTEP, such as statistics and
publications, that can be helpful for researchers to plan their own projects will be explained.

Keywords : fibrous proteins, repetitive sequence, bioinformatics, spider silk, evolution, mechanical property,
KISTEP
References
1. D. Jung, Y. J. Yang, and H. J. Cha, Biotechnol. J. 14, 1900138 (2019).

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2021 KSBB Spring Meeting and International Symposium

[S5-3]
일반특강

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S531 Development of L-Ornithine Hydrochloride and Its Application Process to KFDA as A Food
Additive

Keumsoo HAN, Minhong KIM

MH2 Biochemical Co. Ltd., Korea
Corresponding Author Email : [email protected]

Ornithine is a compound that exists in most living organisms and is a non-protein amino acid. Ornithine is a
proline precursor that is a component of collagen in the skin tissue and is involved in cell growth and
differentiation, which is also involved in the release of ammonia produced during protein metabolism out of the
body. Currently, ornithine is used for the treatment of encephalopathy due to poor liver function and is used as a
special purpose food for baby formula and as a flavor enhancer to offset the bitterness of foods.
The manufacture of Ornithine has begun by Kyowa Hakko in Japan since 1957, and is manufactured by
Ajinomoto and Evonik in Germany using fermentation processes. In 2005, the company manufactured ornithine
by enzymatic processes and exported it mainly abroad ever since. Although it is not yet permitted as a food
additive in Korea, it has been listed in Codex since 2013 and is mainly applied as a flavor enhancer in Japan, the
United States, and Thailand.

In order to be enlisted as a food additive in Korea, food additives draft must be applied to KFDA. The draft must
include the status of enlistment in other foreign countries, development details, manufacturing data, the food
additive specification, technical needs of the additive, human safety, and dosage in foods used for.
In this presentation, I would like to compare the application processes of food additives with foreign countries
and to suggest some consideration of preparing experimental data.

Keywords : L-Ornithine Hydrochloride, Food Additive, Health Benefit, Safety, KFDA

References
1. Evaluation of Certain Food Additives (Seventy-sixth Report of the Joint FAO/WHO Expert Committee on
Food Additives) WHO Technical report Series 974, 2012.
2. Shigeru Ishida, M. Sarada, H. Seki, L. McGirr, A. Lau, K. Morishida, Regulatory Toxicology and
Pharmacology 67, 36 (2013).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S532 Development and Commercialization of Magnetic Force-Assisted Electrochemical Sandwich

Immunoassay (MESIA) for Point-of-Care Testing

Hyundoo HWANG
BBB Inc., Seoul, Korea
Corresponding Author Email : [email protected]

Immunoassay has been one of key technologies for determination of biomarkers in complex biological fluids
such as blood, urine, and saliva. Enzyme-linked immunosorbent assay (ELISA), which is the most widely used
immunoassay technique, is the gold standard for quantitative analysis of protein biomarkers. However, it takes
several hours of analysis, and requires huge and expensive instruments as well as intervention of experts. Rapid
diagnostic test (RDT), which is based on lateral flow immunoassay (LFIA), allows low-cost, rapid detection of
biomarkers. However, it is neither accurate nor precise, thus usually applied for qualitative screening of non-
critical biomarkers. Recently, bench-top immunoassay systems based on microfluidics and fluorescence have
been introduced to the market. They are more precise than RDT, but require expensive optical components
sensitive to dusts and vibration, as well as additional steps with external instruments for sample preparation.
Therefore, the commercial bench-top systems are not appropriate for the point-of-care testing and self-
monitoring of disease markers.
Here we introduce a novel immunoassay technique, called MESIA (magnetic force-assisted electrochemical
sandwich immunoassay). MESIA provides both high accuracy and usability, enabling self-tests at the point of
care with a drop of finger-prick blood. Once a blood sample is loaded into a tiny cartridge, all the processes
from sample preparation to analysis are automatically conducted in a hand-held reader system. The analytical
and clinical performances of MESIA for quantification of blood biomarkers were evaluated according to the
CLSI guidelines. They showed high reproducibility, sensitivity, and specificity as well as excellent agreement
with commercial hospital immunoassay systems. Recently, we developed and commercialized the COVID-19
antigen kit and obtained FDA EUA for the first time among the products made in Korea. MESIA is now
recognized as one of new categories for immunoassay technologies in addition to ELISA and LFIA. In this talk,
we present the history of development and commercialization of MESIA, and discuss the future outlooks of the
new technology.

Keywords : Immunoassay, In vitro diagnostics, Lab-on-a-chip, Point-of-care, Biomarkers

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S533 O2-tolerant Ni–Fe Carbon Monoxide Dehydrogenase by Subsrate Tunnel Engineering

Suk Min KIM, Jinhee LEE, Seung Hyuk KANG, Yong Hwan KIM
Department of Energy and Chemical Engineering, Ulsan National Institute of Science and Technology (UNIST),
Ulsan, Korea
Corresponding Author Email : [email protected]

The extremely O2-sensitive Ni–Fe carbon monoxide dehydrogenase (CODH) is one of the most attractive
enzymes for toxic CO removals from nature and air pollutants [1], since these enzymes can catalyse the oxidation
of CO to CO2 at high rate (>10,000 s-1). To improve O2-tolerance of these CODHs is a big challenge for
biocatalyst use in industrial combustion processes, such as steel mills. Here we showed that the characteristic
change of substrate tunnel close to the active site (C-cluster) of ChCODH-II (Carboxydothermus hydrogeno-
formans), enables to increase O2-tolerance of Ni–Fe CODH. Our experiments revealed that increase of O2-
tolerance compared to that of wild type is achieved for the redesigned ChCODH-II mutant near atmospheric
condition. Moreover, this mutant efficiently catalyzed the oxidation of CO contained in the real waste gases of
LDG (Linz–Donawiz converter Gas) discharged in the steel mill. Our newly designed enzyme will be definitely
a working horse in the removal of toxic gas from environmental pollutants as well as the conversion of waste
CO gas into value-added chemicals.

Keywords : Carbon monoxide, Ni-Fe CO dehydrogenase, Oxygen tolerance, Gas tunnel, Waste gas
References
1. S. W. Ragsdale, Crit. Rev. Biochem. Mol. Biol. 39, 165 (2004).

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2021 KSBB Spring Meeting and International Symposium

S534 Antibacterial Activity of Metal Nanowire and Metal Oxide Thin Films

Ji-Hyeon KIM, Junfei MA, Ga-Hyun LEE, Seunghun LEE, Chang Su KIM
Advanced Nano-Surface Department, Korea Institute of Materials Science, Changwon, Korea
Corresponding Author Email : [email protected]

Zinc oxide (ZnO) and silver nanowire (AgNW), identified as promising antibacterial agents, were investigated
on the antibacterial performance and associated properties. Firstly, to evaluate the functional sustainment of
ZnO antibacterial film in an ozone disinfection system, ZnO sol-gel films were subjected to ultraviolet–ozone
treatment for different periods. Despite the morphological and chemical changes, the satisfactory antibacterial-
activity against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) was sustained even after 2 h
treatment. Secondly, the antibacterial activity of a AgNW spin-coated film were enhanced by 1200 kGy electron
beam irradiation. Silver-oxide islands generated by the irradiation improved the antibacterial activity from 93.5
to 97.3 % for S. aureus and 96.1 to 99.9 % for E. coli. Thirdly, the antibacterial activity of AgNW spin-coated
films was improved by post-heat treatment. AgNW films were heated at various temperatures for 30 min. With
increasing the heating temperature, the AgNWs broke into more segments, and became more oxidized, and
hence the antibacterial activity against E. coli increased from 92.6 to 95.7 %.

Keywords : antibacterial film, zinc oxide, silver nanowire

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S5-4]
From Lab. Research to
Plant Design: Bioprocess
Modeling and Analysis

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S541 Integrated Biorefinery Process from Process Optimization to Process Design for Production of
Value-added Products

Chulhwan PARK
Department of Chemical Engineering, Kwangwoon University, Seoul, Korea
Corresponding Author Email : [email protected]

The biorefinery is well known platform that converts various biomass to value-added products (bioenergy,
biochemical and biomaterial etc.). Various applications such as selection of biomass, optimization of pretreat-
ment, improvement of process efficiency and design of hybrid process can be applied for the sustainable
utilization of biomass. In this presentation, the following four categories will be presented briefly; (1) bioenergy
and biochemical production from lignocellulosic biomass, (2) enzymatic coproduction of value-added products
from vegetable oil, (3) formate ester production based on C1 gas, and (4) process design and evaluation for
value-added chemicals production.

Keywords : biorefinery, biomass, optimization, value-added chemical, process design

References
1. H. Kim et al., Improved production of bacterial cellulose through investigation of effects of inhibitory
compounds from lignocellulosic hydrolysates, Global Change Biology Bioenergy, 13, 3, 436-444 (2021).
2. M. Shin et al., Novel and efficient synthesis of phenethyl formate via enzymatic esterification of formic
acid, Biomolecules, 10(1), 70 (2020).
3. C. Park et al., Eco-design and evaluation for production of 7-aminocephalosporanic acid from carbohydrate
wastes discharged after microalgae-based biodiesel production, Journal of Cleaner Production, 133, 511-517
(2016).
4. J. Joo et al., Improved fermentation of lignocellulosic hydrolysates to 2,3-butanediol through investigation
of effects of inhibitory compounds by Enterobacter aerogenes, Chemical Engineering Journal, 306, 916-924
(2016).

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2021 KSBB Spring Meeting and International Symposium

S542 Development of Biomass Production Process using Ettlia sp. for Operational Expenditure (OPEX)
Reduction

Minsik KIM
Cell Factory Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon,
Korea
Corresponding Author Email : [email protected]

In recent decades, carbon dioxide emission has been striking the globe and carbon-neutral energy sources have
become necessary. Biomass has been spotlighted to solve the greenhouse effect and microalgal biomass
especially is gaining attention with advantages regarding food-ethics and deforestation. In this study, a
cultivation process for a microalga species, namely Ettlia sp. YC001, was statistically optimized. The optimi-
zation under heterotrophy increased biomass and lipid productivities by 4.8-fold and 4.1-fold, respectively.
Subsequently, two feedstocks, i.e., Helianthus tuberosus and algal residue were utilized as alternative nutrient
sources. Furthermore, a sequential hydrolysis process was developed to utilize both feedstocks efficiently. The
sequential hydrolysis process generated a low-cost medium which resulted in almost identical biomass
productivity and increased lipid productivity relative to the statistically optimized medium. Additionally, an
analogous techno-economic analysis for both processes using the optimized medium and low-cost medium was
conducted to examine the effectiveness of the developed processes. The result showed that hydrolysate medium
decreased two-third and three-fourth of required operational expenditures for the optimized media to produce
Ettlia sp. biomass and lipid, respectively. However, the calculation showed that an excessive amount of capital
expenditure was required to utilize microalgal biomass as biofuel, which in turn requires further research on
decreasing capital expenditure.

Keywords : Acid hydrolysis, Algal residue, Biomass, Ettlia sp., Plackett-Burman Design, Response surface
method, Techno-economic analysis
References
1. M. Kim, B.Lee, H. S. Kim, K. Nam, M. Moon, H.-M. Oh*, and Y. K. Chang*, Sci. Rep. 9, 6830 (2019).
2. M. Kim, J. M. Cho, H. S. Kim, H. Lee, H.-M. Oh*, and Y. K. Chang, Energy Convers. Manag. 211, 112769
(2020).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S543 Toward Biomass-derived Biodegradable Plastics: Process Development and Analyses

Wangyun WON
Department of Chemical Engineering, Kyung Hee University, Yongin, Korea
Corresponding Author Email : [email protected]

Lactic acid (LA) is an important bio-based precursor of polylactic acid (PLA), which is biodegradable and
biocompatible plastic that can replace conventional non-biodegradable plastics. In this study, we propose two
strategies for the production of LA, one from lignocellulosic biomass (Strategy A) and the other from sugar
(Strategy B), and conduct integrative analyses such as techno-economic analysis, pioneer plant analysis,
uncertainty analysis, and life-cycle assessment to investigate the comprehensive feasibility and sustainability for
the proposed strategies. To reduce utility consumption, we perform heat integration. The minimum selling prices
of LA, which are defined as a selling price at breakeven point, for the strategies A and B are estimated to be
$1,498/ton and $1,491/ton, respectively, which are lower than current market price of LA, i.e., $1,526/ton,
indicating that LA produced from the proposed strategies is cost-competitive. These MSPs can increase to
$2,340/ton and $1,641/ton for strategies A and B, respectively, under the assumption of pioneer plant. The
uncertainty analysis using Monte-Carlo simulation shows risk and uncertainty in our new strategies by means of
cumulative probability and frequency graphs. Through the life-cycle assessment, we quantify the environmental
impact of the proposed process.

Keywords : biodegradable, economics, sustainability

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S544 Process Design and Assessment of Electrochemical Carbon and Nitrogen Reduction Processes

Juyeon KIM, Jonggeol NA

Department of Chemical Engineering and Materials Science, Ewha Womans University, Seoul, Korea
Corresponding Author Email : [email protected]

Electrochemical processes–converting carbon dioxide and nitrogen to value-added precursors such as ethylene
and ammonia–are promising techniques to escape from the petrochemical-dependent industrial structure.
However, assessments of the economics of overall processes simultaneously are difficult due to complicated
reaction networks and product combinations affecting optimal process design. Here, we introduce a general
techno-economic analysis of electrochemical carbon dioxide and nitrogen reduction processes through a
superstructure-based automated process synthesis platform and thereby considering the overall process design
combinations simultaneously, which leads to the rigorous assessment. The economical global sensitivity
according to current density, Faraday efficiency, and overpotential is evaluated. The analysis highlights the
promise optimal design for 1) coupled process of carbon dioxide reduction with organic oxidation reaction, and
2) nitrate production through electrochemical nitrogen oxidation and ammonia production through nitrate
reduction in a single reaction system.

Keywords : electrochemical, nitrogen reduction, carbon dioxide reduction, carbon capture and utilization,
process systems engineering, techno-economic analysis
References
1. Na, J., Seo, B., Kim, J. et al., Nat Commun. 10, 5193 (2019).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S5-5]
Recent Advances of
Electrochemical Biotechnology
for Energy and Environment

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S551 Nanobiocatalysis for CO2 Conversion and Utilization

Jungbae KIM
Department of Chemical and Biological Engineering, Korea University, Seoul, Korea
Corresponding Author Email : [email protected]

Even though enzymes can be employed and newly proposed for various applications such as electrochemical
biosensors, membrane antifouling and CO2 conversion, their successes in practical applications are often
hampered due to their poor stability. Nanobiocatalysis, immobilizing enzymes using various nanomaterials, has
demonstrated its successes in stabilizing the enzyme activity for various enzyme applications such as biosensors,
biofuel cells, enzyme-linked immunosorbent assay, membrane antifouling, and CO2 conversion. This presenta-
tion will mainly cover nanobiocatalytic stabilization of carbonic anhydrases for their potential successes in CO2
conversion. Especially, the nanobiocatalytic stabilization of carbonic anhydrase, catalyzing the hydration of CO2
to bicarbonate at a 105-6 turnover number, has made an unprecedented success by maintaining 83% of initial
enzyme activity after incubation in aqueous solution under shaking at 200 rpm for two years. Stabilized carbonic
anhydrase was successfully employed for the effective conversion of CO2 to bicarbonate, which was further
used for expedited microalgae growth and improved calcium carbonate production. If time permits, several
other examples of nanobiocatalytic applications together with several other research areas will be introduced.

Keywords : Nanobiocatalysis, carbonic anhydrase, CO2 conversion, expedited microalgae growth, calcium
carbonate formation
References
1. Han Sol Kim, Han Sol Kim, Sung-Gil Hong, Kie Moon Woo, Vanesa Teijeiro Seijas, Seongbeen Kim,
Jinwoo Lee and Jungbae Kim, ACS Catal. 8, 6526-6536 (2018).
2. Han Sol Kim, Sung-Gil Hong, Jusang Yang, Youngjun Ju, Joongbok Ok, Seok-Joon Kwon, Kyung-Min
Yeon, Jonathan S. Dordick, Jungbae Kim, J. CO2 Util., 38, 291-298 (2020).
3. Seung-Hyun Jun, Jusang Yang, Hancheol Jeon, Han Sol Kim, Seung Pil Pack, EonSeon Jin, and Jungbae
Kim, Environ. Sci. Technol., 54, 1223-1231 (2020).

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2021 KSBB Spring Meeting and International Symposium

S552 Electrochemical Biosensors Using Redox Cycling

Haesik YANG
Department of Chemistry, Pusan National University, Busan, Korea
Corresponding Author Email : [email protected]

High signal amplification is essential for ultrasensitive detection of biomolecules. In recent years, we have
developed new redox cycling schemes, combined with enzymatic or nanozymatic amplification, that allow for
high signal amplification in a simple format along with low background levels. To minimize unwanted side
reactions, all involved species were properly selected. In this presentation, the basic concept of the developed
redox-cycling schemes will be explained, and their application to the detection of microorganisms including
Escherichia coli (E. coli) and Aspergillus niger (A. niger) will be introduced. The E. coli detection method
harnesses the intracellular β-galactosidase (Gal) activity of E. coli along with the signal amplification based on
redox cycling. The Gal expression level is increased by treatment with a Gal expression-inducer; the enzymatic
reaction of Gal is facilitated by the permeabilization treatment involving the use of chloroform and sodium
dodecyl sulfate; the electrochemical signal is amplified by the electrochemical–chemical–chemical and
chemical–chemical redox cycling involving the Gal product. A sensitive detection method specific to A. niger is
based on a single-mediator system combined with electrochemical-chemical redox cycling. Intracellular
NAD(P)H-oxidizing enzymes in A. niger converts electro-inactive 4-nitro-1-naphthol into electro-active 4-
amino-1-naphthol. Since the membrane and wall of A. niger is well permeable to both 4-nitro-1-naphthol and 4-
amino-1-naphthol in tris buffer (pH 7.5) solution, the electrochemical signal is increased in the presence of A.
niger.

Keywords : Biosensors, Escherichia coli, Aspergillus niger

References
1. H. Yang, Curr. Opin. Chem. Biol. 16, 422 (2012).
2. S. Noh, Y. Choe, V. Tamilavan, M. H. Hyun, H. Y. Kang, H. Yang, Sens. Actuators B: Chemical 209, 951
(2015).
3. J. Kwon, E.-M. Cho, P. Nandhakumar, S. I. Yang, H. Yang, Anal. Chem. 90, 13941 (2018).
4. P. Nandhakumar, G. Kim, S. Park, S. Kim, S. Kim, J. K. Park, N.-S. Lee, Y. H. Yoon, H. Yang, Angew.
Chem. Int. Ed. 59, 22419 (2020).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S553 Synthetic Analogs of Mo/W Enzymes and Reactivity with CO2 and Protons

Junhyeok SEO
Department of Chemistry, Gwangju Institute of Science and Technology, Gwangju, Korea
Corresponding Author Email : [email protected]

Synthetic modeling of metalloenzyme active sites can provide understandings of enzyme reactivity, as well as
research insights into the development of catalysts for analogous reactions. We are interested in transition metal
complexes having CO2-conversion reactivity, and as a research approach, we study synthetic analogs of formate
dehydrogenases (FDHs) active sites. In some prokaryotes, the FDHs, containing molybdenum (Mo) or tungsten
(W) cofactors, facilitate the reversible conversion of formate/CO2 (HCO2– ⇄ CO2 + H+ + 2e–). Remarkably, the
enzyme operates near the thermodynamic potential of the reduction of CO2 to formate (–0.43 V vs. NHE at pH =
7). The FDH active site has a high valent Mo/W(IV,VI) center coordinated by equatorial bis-dithiolene and axial
chalcogenide ligands. The dithiolene structure plays a role as an electron sink to accept/donate electrons from/to
the metal ions while stabilizing the high valent reaction core, and the chalcogenide ligand determines the
reactivity with substrates.
A synthetic model, [WIV(OH)(S2C2Ph2)2]– complex, where a hydroxy (OH) group coordinates to the W ion in the
axial position, reacted with CO2 in the fashion of O-atom exchange between W-OH/CO2 through a [W-CO3H]
intermediate. The nucleophilic hydroxide group seems to initiate the reaction, however, the next step to the
formate generation was not achieved by the current model. On the other hand, we investigated the CO2 reactivity
of a thermally stable [WIV(O)(S2C2Me2)2]2–, which showed stability at ~100 °C under N2 atmosphere, but
interestingly, the W(IV) ion was oxidized to W(V) while reacting with CO2, and the complex was structurally
reorganized to a triply bridged dinuclear W(V) complex, [W2O2(μ-S)(μ-S2C2Me2)(S2C2Me2)2]2–. During the
oxidative conversion, the stoichiometric amount of CO2 was reduced to formate in the presence of a proton
source. In this talk, I will discuss the experimental data and current understandings of the (electro)chemical
reactivities of the Mo/W-bis(dithiolene) complexes.

Keywords : formate dehydrogenase, synthetic analogs, Mo/W dithiolene complexes

References
1. Seo. J.*, Shearer, J.; Williard, P. G.; Kim, E.*, Dalton Trans., 48, 17441 (2019).

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2021 KSBB Spring Meeting and International Symposium

S554 Prevention of Electrochemical Degradation in Bioelectrochemical System by Electrical Control

Taeyoung KIM
Department of Environmental Engineering, Chosun University, Gwangju, Korea
Corresponding Author Email : [email protected]

The bioelectrochemical system is recognized for its potential as a useful future environmental-energy techno-
logy in terms of sustainable wastewater treatment and green energy production1,2). Though the bioelectroche-
mical technology has rapidly developed over the past 20 years, until now, the scale-up of the bioelectrochemical
system to the level of industrial application in the real world has not been achieved. The problems in
bioelectrochemical degradation such as voltage reversal phenomenon and power overshoot should be overcome
in order to achieve a proven level of wastewater treatment efficiency and power generation3). In this study, three
multi-electrode-embedded microbial fuel cells (MFCs) were sequentially connected and operated in series and
parallel modes, fed by effleunt of an anaerobic digester continuously operated using swine wastewater. The
MFCs connected in parallel showed the higher power density compared to the series mode. The bioelectroche-
mical degradations were prevented by MFCs connected in parallel and connected with power management
system despite inevitable electrical malfunction conditions by imbalance organic loadings. These findings can
lead to be closer real application of bioelectrochemical systems for sustainable wastewater treatment and
bioenergy generation.

Keywords : bioelectrochemical system, microbial fuel cell, wastewater treatment, voltage reversal, power
management system
References
1. Bruce E. Logan, Bert Hamelers, René Rozendal, Uwe Schröder, Jürg Keller, Stefano Freguia, Peter
Aelterman, Willy Verstraete, and Korneel Rabaey, Microbial fuel cells: methodology and technology
(2006), Environmental Science & Technology, 40, 5181-5192.
2. René A. Rozendal, Hubertus V.M. Hamelers, Korneel Rabaey, Jurg Keller, Cees J.N. Buisman, Towards
practical implementation of bioelectrochemical wastewater treatmetn(2008), 26, 450-459.
3. Bongkyu Kim, S. Venkata Mohan, Deby Fapyane, In Seop Chang, Controlling voltage reversal in microbial
fuel cells(2020), Trends in Biotechnology, 38(6), 667-678.

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2021 KSBB Spring Meeting and International Symposium

[S5-7]
Frontiers in C1 Gas Refinery

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S571 Methanotrophic Platform Cell Factory for Methane Bioconversion

Eun Yeol LEE

Department of Chemical Engineering, Kyung Hee University, Yongin, Korea
Corresponding Author Email : [email protected]

Methane is a low-priced next-generation carbon feedstock for industrial biotechnology. Methanotrophs can
convert methane to value-added products. Methane bioconversion using methanotrophic platform cell factory
can valorize methane and mitigate methane as greenhouse gas. With recent advances in molecular biology-based
tool development and system biology-based understanding of methanotrophs’ physiology, methanotrophs can be
engineered to produce various target products from methane. In this presentation, technical challenges and
issues of methanotrophs as a platform cell factory for methane bioconversion will be discussed.

Keywords : Methanotrophs, methane, bioconversion

References
1. Anh Duc Nguyen, Eun Yeol Lee, Engineered methanotrophy: a sustainable solution for methane-based
industrial biomanufacturing (2021). Trend. Biotechnol. doi: 10.1016/j.tibtech.2020.07.007
2. Diep Thi Ngoc Nguyen, Ok Kyung Lee, Nguyen Thi Thu and Eun Yeol Lee, Type II methanotrophs: a
promising microbial cell-factory platform for bioconversion of methane to chemicals (2021). Biotechnol.
Adv., 47, 107700.

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2021 KSBB Spring Meeting and International Symposium

S572 Synthetic Microbial Consortium for the Production of High Value Chemicals from Carbon
Monoxide

Gyoo Yeol JUNG1,2

1
School of Interdisciplinary Bioscience and Bioengineering, POSTECH, Pohang, Korea,
2
Department of Chemical Engineering, POSTECH, Pohang, Korea
Corresponding Author Email : [email protected]

Carbon monoxide is produced from iron smelting as a byproduct and a large quantity of CO byproduct is also
formed during the oxidative processes for the production of chemicals. For this reason, it is important to develop
a process for the conversion of CO to high value chemicals such as bioactive compounds, animal feeds, and
platform chemicals. Biological process is known as the most efficient method to transform from CO to high
value products because some microorganisms can grow by using CO as sole carbon source. Naturally available
CO consuming microorganisms are, however, hard to be engineered due to lack of genetic tools and their
biological information. In this presentation, synthetic microbial consortium using novel synthetic biology tools
for the development of efficient biological process converting CO to high value chemicals.

Keywords : Microbial consortia, Carbon monoxide, Fermentation stability, 3-Hydroxypropionic acid, Itaconic
acid
References
1. Sanghak Cha et al, Design of mutualistic microbial consortia for stable conversion of carbon monoxide to
value-added chemicals (2021), Metabolic Engineering, 64, 146-153.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S573 Biotransformation of Formaldehyde into Glycolic Acid, Ethylene Glycol, and Ethanol Amine via
Glycolaldehyde by using Glyoxylate Carboligase as a Key Enzyme

Jeong-Sun KIM1, Jin-Byung PARK2

1
Department of Chemistry, Chonnam National University, Gwangju, Korea, 2Dapartment of Food science and
Engineering, Ewha Womans University, Seoul, Korea
Corresponding Author Email : [email protected]

Formaldehyde (HCHO) is an emerging C1 source, because it could be prepared from CH4, CH3OH, CO, and
CO2 by biological and/or chemical means [1]. A very simple biocatalytic system for the synthesis of the
industrially relevant C2 chemicals (e.g., glycolic acid, ethylene glycol, and ethanol amine [2]) from formal-
dehyde was established. The biocatalytic system consisted of a newly discovered thermostable glyoxylate
carboligase from Escherichia coli K-12 (EcGCL) and an aldehyde dehydrogenase (e.g., α-ketoglutaric
semialdehyde dehydrogenase (KGSADH) from Azospirillum brasilense) or an alcohol dehydrogenase (i.e., 1,3-
propanediol dehydrogenase (DhaT) of Klebsiella pneumoniae) or ω-transaminase (ω-TA) from Silicibacter
pomeroyi [2]. The combination of EcGCL and KGSADH allowed to produce glycolic acid from formaldehyde
via glycolaldehyde to a conversion of 62%, while the serial reaction of EcGCL and DhaT led to formation of
ethylene glycol to a conversion of 64%. The combination of EcGCL and ω-TA enabled the production of
ethanol amine to a conversion of 64%. This study will contribute to enzymatic synthesis of glycolic acid,
ethylene glycol, and ethanol amine from C1 compounds in an environment-friendly way.

Keywords : Formaldehyde, Glyoxylate carboligase, Glycolaldehyde

References
1. Sarah Desmons, Régis Fauré, Sébastien Bontemps, Formaldehyde as a promising C1 source: the instru-
mental role of biocatalysis for stereocontrolled reactions (2019) ACS Catalysis, 9, 9575-9588.
2. Laura Salusjärvi, Sami Havukainen, Outi Koivistoinen, Mervi Toivari, Biotechnological production of
glycolic acid and ethylene glycol: current state and perspectives (2019) Applied Microbiology and Bio-
technology, 103, 2525-2535.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S574 Systems Biology of Acetogenic Bacteria for Sustainable Bioproduction from C1 Feedstocks

Byung-Kwan CHO
Department of Biological Sciences, KAIST, Dajeon, Korea
Corresponding Author Email : [email protected]

The C1 gases generated in various industrial processes consist of carbon dioxide (CO2), carbon monoxide (CO),
and methane (CH4). Because these gases cause global climate change, such as the greenhouse effect and
environmental pollution, acetogenic bacteria (acetogens) have emerged as a biocatalyst to recycle C1 gases by
converting them into value-added biochemicals using the Wood-Ljungdahl (WL) pathway. However, despite the
advantage of applying acetogens as biocatalysts, lack of systemic understanding such as the regulation
mechanism of the WL pathway or the transfer process of an external electron to fix C1 compounds has limited
the construction of a cellular factory optimized. Instead, most previous studies focused on understanding the
biochemistry of WL pathway and physiological change of acetogen. To overcome the limitation of lack of
knowledge about the genetic landscape, we have considered understanding the C1 fixation mechanism in
acetogen at the molecular-levels based on the system biology. Through this approach, we understand the WL
pathway's regulation mechanism, discovered a novel C1 fixation pathway, or demonstrated how the redox
energy generated from the external electrode could be utilized to fix the C1 gas. These research results will
suggest the direction of strain improvement necessary for constructing an efficient C1 gas fixing system using
acetogen.

Keywords : Acetogenic bacteria, Wood-Ljundahl pathway, System biology

References
1. Y. Song et al., Functional cooperation of the glycine synthase-reductase and Wood–Ljungdahl pathways for
autotrophic growth of Clostridium drakei (2020), PNAS, 117(13).
2. S. Jin et al., Acetogenic bacteria utilize light-driven electrons as an energy source for autotrophic growth
(2021), PNAS, 118(9).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

부문위원회 및 학생세션

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S6-1]
부문위원회 및 학생세션 I:
Fresh Ph.D.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6101 A Novel CO2-free Approach for the pH-control and Recycling of Bicarbonate-carbonate-
buffered Culture Media in Microalgae Production Systems

Niklas ZELL1, Sunhwa JUNG1, Fabian BOßLE1, Marc HERDEN1, Peter KURZWEIL1, Franz BISCHOF1,
Aelxander JAHN2, Chrsitoph LINDENBERGER1
1
OTH Amberg-Weidern, Department of Mechanical Engineering and Environmental Engineering, Amberg,
Germany, 2German Engineering Research and Development Center LSTME Busan, Busan, Korea
Corresponding Author Email : [email protected]

Over the past decades, microalgae have developed into a promising source of biofuels and -materials as well as
a possible carbon sink for carbon capture technologies and yet their cultivation still poses economic challenges,
especially regarding their carbon supply. Owing its simplicity, the cultivation in highly saline bicarbonate-
carbonate-buffered media is an appealing way to address this problem, but only as long as the media’s pH can
be kept low enough to avoid the shift of available bicarbonate into worthless carbonate. Most methods proposed
for such a pH-control rely on the use of CO2- or flue-gas, which is limited by the relatively low mass transfer.
Therefore, a different approach is presented, that is based upon perm-selective cation-exchange-membranes and
completely omits the use of gaseous CO2-supply for pH-control. During the process, the cultivation media flows
over the membrane that separates the broth from an acid reservoir (here hydrochloric acid). The H+ from the acid
pass through the membrane, lowering the broth’s pH without the introduction of additional anions. The transfer
of Na+ and K+ from the media to the acid balances the electrical flow. Using a lab-scale membrane-module, this
method was applied to the used cultivation media of Arthrospira platensis cultures, which has risen to pH 12,
resulting in a low vitality of the culture. After lowering the pH back to 8 again, thereby shifting all the
carbonates back to available bicarbonates, the medium was used for a second batch culture of A. platensis. The
growth in the regenerated media coincided well with that of the cultures fed with fresh medium, thus clearly
demonstrating the effectiveness of this method in terms of pH-control, overall medium-recycling and improved
substrate-use efficiency.

Keywords : Microalgae, cation exchange membrane, pH control, CO2 free, media regeneration

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6102 Numerical Investigation of Fluid Flow, Light Distribution and Cellular Growth Kinetics in
Bubble Column Photobioreactors

Giovanni LUZI1, Christopher MCHARDY2, Bastian EYSEL2, Christoph LINDENBERGER3,

Cornelia RAUH1,2,4, Antonio DELGADO1,4
1
LSTME Busan Branch, Busan, Korea, 2Department of Food Biotechnology and Food Process Engineering,
Technical University of Berlin, Berlin, Germany, 3Department of Mechanical Engineering and Environmental
Engineering, OTH Amberg, Kaiser-Wilhelm-Ring, Amberg, Germany, 4Institute of Fluid Mechanics, University
of Erlangen-Nuremberg, Cauerstraße, Erlangen, Germany
Corresponding Author Email : [email protected]

One of the most promising potential choices for sustainable production of food, feed, or fuels is represented by
microalgae. These microorganisms are cultivated in photobioreactors (PBR) where the maximum culture density
is limited to few grams per liters. To increase the PBR productivity, researchers suggest promoting flashing light
conditions, either by intensive mixing or by applying illumination strategies like light pulsation. Independently
of the chosen approach, the light distribution represents a critical parameter for the growth of phototrophic
microorganisms, and it is mostly affected by the microorganism concentration and by the distribution of gas
bubbles.
Utilizing numerical simulations, we first investigated the influence of gas bubbles on the light field at different
gas superficial velocities, concluding that the presence of gas bubbles affects both the spatial distribution and the
magnitude of the light intensity field only at biomass concentrations less than 1 kg/m3, but generally to a very
small extent. Afterward, we compared pneumatic mixing and flashing LED sources regarding their feasibility to
promote the flashing light effect. On the one hand, we found that at the investigated operating conditions
pneumatic mixing alone has no impact on the growth rate and a further increase of the mixing intensity might be
limited by the shear sensitivity of the cells. On the other hand, numerical simulations clearly show that
illumination with flashing LED promotes an increase of the growth rate up to a factor of 2.5 if proper flashing
frequencies are selected. Our numerical simulations integrate the computation of the three-dimensional fluid
flow and light field, as well as the computation of the growth kinetics of algal cells. In our studies, we selected a
5 cm diameter bubble column PBR filled with a suspension of Chlamydomonas reinhardtii, which is a reactor
size relevant for industrial conditions.

Keywords : Bubble Column, Numerical Simulations, Flashing LED, Growth Kinetics

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6103 Development of an in vitro Gut Simulation Device for the Investigation of Host-microbe
Interaction by Integrative Multiomics Analysis

Won-Suk SONG, Sung-Hyun JO, Jae-Seung LEE, Hyo-Jin JEON, Ji-Eun KWON, Ji-Hyeon PARK,
Ye-Rim KIM, Yun-Gon KIM
Department of Chemical Engineering, Soongsil University, Seoul, Korea
Corresponding Author Email : [email protected]

The commensal gut bacterium Akkermansia muciniphila is in the spotlight as a promising probiotic candidate
that improves host health and prevents diseases. However, the biological interaction of A. muciniphila with
human gut epithelial cells has rarely been explored for use in biotherapeutics. Here, this study elaborates on the
development of the in vitro device that simulates the gut epithelium to elucidate the biological effects of living
anaerobic bacteria via multiomics analysis: the Mimetic Intestinal host-Microbe Interaction Coculture System
(MIMICS). Both human gut epithelial cells (Caco-2) and the anaerobic bacterium, Akkermansia muciniphila,
can remain viable for 12 hours after coculture in the MIMICS. The changes of transcriptomics and proteomics
(i.e. cell-cell junctions, immune responses, and mucin secretion) in gut epithelial cells after the treatment with A.
muciniphila closely correspond with those reported in previous animal studies. In addition, multiomics results
revealed that A. muciniphila activates glucose and lipid metabolism in gut epithelial cells, leading to the increase
in ATP production. This study suggests that the MIMICS may be an effective general tool for evaluating the
effects of anaerobic bacteria on gut epithelial cells.

Keywords : Akkermansia muciniphila, Host-microbe interaction, multiomics analysis, gut simulation device,
LC-MS/MS
References
1. W. Song et al, Biotechnol. Bioeng. Online published (2021).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6104 Gold Nanoparticle-assisted SELEX as a Visual Monitoring Platform for Rapid Discovery of
Small Molecule-binding DNA Aptamers

Eun-Song LEE1,2, Jeong Min LEE1, Hea-Jin KIM1, Gyeongmin KIM1, Young-Pil KIM1,2
1
Department of Life Science, Hanyang University, Seoul, Korea, 2Research Institute for Convergence of Basic
Sciences, Hanyang University, Seoul, Korea
Corresponding Author Email : [email protected]

To resolve time-consuming and imperceptible monitoring problems in the traditional systematic evolution of
ligands by exponential enrichment (SELEX), we report gold nanoparticle-assisted SELEX (GNP-SELEX) as a
visual and straightforward monitoring platform for the rapid discovery of small molecule-binding single-
stranded DNA (ssDNA) aptamers. Through the colorimetric changes within and between rounds, GNP-SELEX
enabled the rapid determination of target-specific aptamer library enrichment with neither target modification
nor post-SELEX monitoring process. We also identified ssDNA aptamers with high selectivity and binding
affinity by targeting two hormones (brassinolide; BL and bisphenol A; BPA) as a model. The rational design of
selected aptamers by 3D molecular simulation increased their ability to detect BL or BPA in real samples as
bioreceptors. These results suggest that GNP-SELEX is directly useful as a self-monitoring platform to discover
ssDNA aptamers for diverse targets in a rapid and simple way.

Keywords : Gold Nanoparticle, Aptamer, SELEX, Brassinolide, Bisphenol A

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6105 Local Therapeutic Nanoplatform Based on Stimuli-responsive Mussel Adhesive Protein For
Highly Efficient Cancer Therapy

Yeonsu JEONG1, Yun Kee JO2, Hyung Joon CHA1

1
Biomaterials Research Center, Pohang University of Science and Technology (POSTECH), Pohang, Korea,
2
Department of Biomedical Convergence Science and Technology, Kyungpook National University, Daegu, Korea
Corresponding Author Email : [email protected]

Nanoparticles (NPs)-based drug delivery system has been developed with advantages of passive targeting ability
and controlled release property, however, systemic approaches have several limitations such as insufficient
accumulation and systemic toxicity. In addition, harsh microenvironments including flow of body fluids have
been major challenges in effective bioavailability of therapeutics. Thus, local therapeutic system requires
innovative NPs with great adhesion, stimuli-sensitivity, and high biocompatibility to improve therapeutic
absorption. Here, we propose mussel adhesive protein (MAP)-based pH- and photo-responsive NPs to provide
prolonged retention on tumor tissue and facilitate the controlled release of therapeutics for local cancer therapy.
The MAP NPs showed superior surface adhesion and selective drug release properties responding to pH and
light, enabling localized chemo-, gas-, and photothermal-therapy. The locally-treated MAP NPs exhibited
significant anticancer effects in vivo devoted by extended retention on tumor sites. We anticipate that the MAP-
based nanoplatform can offer the versatile approaches for effective cancer therapy with few risk of adverse
effects.

Keywords : Mussel adhesive protein, Adhesive nanoparticle, Stimuli-responsive nanoplatform, Local drug
delivery
References
1. Y. Jeong, Y. K. Jo, B. J. Kim, B. Yang, K. I. Joo, and H. J. Cha, ACS Nano 12, 8909 (2018).
2. Y. Wang, Y. Deng, H. Luo, A. Zhu, H. Ke, H. Yang, and H. Chen, ACS Nano 11, 12134 (2017).
3. K. Park, ACS Nano 7, 7442 (2013).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6106 A Novel Bicelle Preparation Method Using Microfluidic Hydrodynamic Focusing

Sunghak CHOI1, Bongsu KANG2, Keesung KIM3, Ho-Sup JUNG1

1
Dept. of Food Science and Biotechnology, Center for Food and Bioconvergence, Seoul National University,
Seoul, Korea, 2Dept. of Mechanical Engineering, Kyungpook National University, Daegu, Korea, 3College of
Engineering, Research Institute of Advanced Materials, Seoul National University, Seoul, Korea
Corresponding Author Email : [email protected]

A bilayered micelle or so-called bicelle offers a variety of potential applications in the biomedical field due to
its high stability and similarity with biological membranes [1]. Thin-film hydration method which is the
conventional method to prepare bicelles demands complicated manufacturing processes so that enhancing
productivity is a significant issue. Moreover, it accompanies harsh conditions such as repeating freezing and
thawing cycles, in which the physicochemical characteristics of functional materials could be denatured [2].
Here, we propose a novel method for bicelle synthesis using a microfluidic chip without any external energy
exertion. Bicelles were successfully prepared using this continuous method, with enhanced physicochemical
properties. The critical condition for bicelle formation in this microfluidic system is deduced through
experimental and analytical studies in terms of concentration and mixing time. Furthermore, the size of bicelle
can be controlled, which crucially influences cellular uptake, transportation, and accumulation behavior.

Keywords : Bicelle, Microfluidic Chip, Hydrodynamic Focusing

References
1. M. Beaugrand, A. A. Arnold, J. Hénin, D. E. Warschawski, P. T. F. Williamson, and I. Marcotte, Langmuir.
30, 6162-6170 (2014).
2. S. Taguchi, K. Suga, K. Hayashi, Y. Okamoto,H.-S. Jung, H. Nakamura, H. Umakoshi, Colloids and
Interfaces. 2, 73 (2013).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6107 Glycan Chip Based on i-motif DNA Linker with pH-responsive Structural Change for On-
surface Biosynthesis of Cancer-associated Glycans

Hye Ryoung HEO1, Jeong Hyun SEO2, Chang Sup KIM3, Hyung Joon CHA1
1
Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Korea, 2School
of Chemical Engineering, Yeungnam University, Gyeongsan, Korea, 3School of Chemistry and Biochemistry,
Yeungnam University, Gyeongsan, Korea
Corresponding Author Email : [email protected], [email protected]

On-chip glycan biosynthesis is an effective strategy for preparing useful complex glycan sources and for
applying glycan-involved applications simultaneously. However, current methods cannot provide quantitative
information on synthesized glycans, resulting in undefined structures on a surface, which leads to unequable and
unreliable results. In this work, novel glycan chip was developed by introducing pH-responsive i-motif DNA
linker capable of reversibly controlling the immobilization and separation of glycans on chip surface in pH-
dependent manner. On-chip enzymatic glycosylations were optimized for synthesizing cancer-associated
glycans such as Globo H hexasaccharide by analyzing quantitatively products isolated from the surface. This
enabled to construct glycan chip composed of structurally defined cancer-associated glycans. Interaction
analysis of anti-Globo H antibody and Globo H-related glycans biosynthesized on the chip showed that VK9
strongly interacts with biosynthesized Globo H hexasaccharide. Furthermore, we investigated glycan-binding
specificity of MCF-7 breast cancer cells using glycan chip consisting of biosynthesized Globo H hexasaccharide
and its related glycans to identify a high-affinity glycan ligand. The cancer cell strongly recognized Globo H
hexasaccharide, suggesting that the specific receptor for Globo H hexasaccharide can be present on the surface
of breast cancer cell. These results demonstrated the feasibility of the DNA linker-based glycan chip for on-
surface complex glycan biosynthesis and glycan-involved applications.

Keywords : glycan chip, on-chip synthesis, enzymatic glycosylation

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S6-2]
부문위원회 및 학생세션 II:
Fresh Ph.D.

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6201 Amino Acids Detection Using a Complementary Cell-free Protein Synthesis System

Hye Jin LIM, Kyung-Ho LEE, Dong-Myung KIM

Department of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, Korea
Corresponding Author Email : [email protected]

Amino acids are building blocks of protein synthesis, and also play important roles in maintaining the
physiological homeostasis. For example, changes in amino acid concentrations are closely related with cell
signaling, gene expression, and protein phosphorylation cascade. In addition, many amino acids serve as the
precursors of key metabolites, including hormones and polyamines. Therefore, quantification of amino acids can
provide important information for disease diagnosis. The detection and quantification of amino acids have
conventionally employed chromatography technologies, including high performance liquid chromatography
(HPLC), capillary electrophoresis (CE), and gas chromatography (GC). However, these methods require
complicated equipment setup, highly trained operator, and long process time.
In this study, we developed a rapid and cost-effective assay method that harnesses the molecular process of
protein synthesis for quantitative detection of amino acids. In this scheme, dubbed complementary cell-free
protein synthesis (CCFPS) assay, a reaction mixture for cell-free protein synthesis is prepared without amino
acids that need to be analyzed. When programmed with a reporter gene, and mixed with an assay sample
containing the missing amino acids, the reaction mixture for this CCFPS assay produces detection signal. With
different reporter enzymes, the intensity of the signal from the CCFPS assay showed linear correlations with the
concentrations of amino acids in assay samples. Furthermore, we could harness the endogenous enzymes of cell
extract for the measurement of non-proteinogenic amino acids by the CCFPS assay.

Keywords : Cell-free protein synthesis, Amino acids, Amino acid analysis, Diagnosis
References
1. M. C. Waldhier, M. A. Gruber, K. Dettmer, and P. J. Oefner, Anal. Bioanal. Chem. 394, 695 (2009).
2. Y. J. Jang, K. H. Lee, T. H. Yoo, D. M. Kim (2017) Anal. Chem., 89, 9638-9642.
3. Y. J. Jang, K. H. Lee, T. H. Yoo, D. M. Kim (2019) Anal. Chem., 91, 2531-2535.
4. Y. J. Jang, K. H. Lee, T. H. Yoo, D. M. Kim 2020) Anal. Chem., 92, 11505-11510.

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2021 KSBB Spring Meeting and International Symposium

S6202 Resolving the Mutually Exclusive Immune Activities of Chitosan

Suyoung LEE1, Seohyun BYUN2, Sin-Hyeog IM2, Dong Soo HWANG1

1
Division of Environmental Science and Engineering, POSTECH, Pohang, Korea, 2Division of Integrative
Bioscience and Biotechnology, Department of Life Science, POSTECH, Pohang, Korea
Corresponding Author Email : [email protected]

Chitosan has multifaceted functions displayed by both pro- and anti-inflammatory properties which hinder its
effective development as an immunomodulatory agent. Herein, the contributions of molecular weight (MW) of
chitosan with regard to its immune regulatory properties towards inflammation are investigated. The anti-
inflammatory properties of low and relatively high MW chitosan are compared by utilizing immunological
assays and nanomechanics based experiments on the surface forces apparatus (SFA). Interestingly, compared
with relatively high MW chitosan, low MW chitosan significantly increases the differentiation of anti-
inflammatory regulatory T cells (Tregs) through the Dectin-1-dependent pattern recognition receptor (PRR) on
antigen-presenting cells in mice. SFA analyses also show a similar trend of interaction forces between chitosan
and diverse PRRs depending on its MW. The results obtained in the immunological and nanomechanical
experiments are consistent and imply that the binding features of PRRs vary depending on the MW of chitosan,
which may alter immune activity. In accordance, in vivo administration of only low and not high MW chitosan,
repress inflammatory responses and suppress the progression of experimental colitis. This study elucidates a
previously unexplored size-dependent immunoregulatory property of chitosan, and suggests the applicability of
low MW chitosan as a pharmaceutical ingredient to treat diverse inflammatory disorders.

Keywords : chitosan, immune activities, pattern recognition receptors, anti-inflammation

References
1. C. L. Bueter, C. K. Lee, J. P. Wang, G. R. Ostroff, C. A. Specht, S. M. Levitz, Journal of immunology, 192,
5943 (2014).
2. K. Azuma, T. Osaki, S. Kurozumi, M. Kiyose, T. Tsuka, Y. Murahata, T. Imagawa, N. Itoh, S. Minami, K.
Sato, Y. Okamoto, Carbohydrate polymers, 115, 448 (2015).
3. T. A. Reese, H. E. Liang, A. M. Tager, A. D. Luster, N. Van Rooijen, D. Voehringer, R. M. Locksley,
Nature, 447, 92 (2007).
4. Carla A. Da Silva, Dominik Hartl, Wei Liu, Chun G. Lee, Jack A. Elias, J Immunol. 181, 4279 (2008).

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2021 KSBB Spring Meeting and International Symposium

S6203 Genome-scale Analysis of Acetobacterium woodii Identifies Translational Regulation of

Acetogenesis

Jongoh SHIN, Yoseb SONG, Seulgi KANG, Sangrak JIN, Sun Chang KIM, Suhyung CHO, Byung-Kwan CHO
Department of Biological Sciences and KI for the BioCentury, Korea Advanced Institute of Science and
Technology, Daejeon, Korea
Corresponding Author Email : [email protected]

Acetogens synthesise acetyl-CoA via the CO2-fixing Wood–Ljungdahl pathway. Despite their ecological and
biotechnological importance, their translational regulation of carbon and energy metabolisms remains unclear.
Here, we report how carbon metabolism in the model acetogen Acetobacterium woodii are translationally
controlled under different growth conditions. In particular, Wood-Ljungdahl pathway genes are translated at
similar levels to achieve efficient acetogenesis activity under autotrophic growth conditions, whereas the
carbonyl-branch presents an increased translation level in comparison to the methyl-branch under heterotrophic
growth conditions. The translation efficiency of genes in these pathways is differentially regulated by 5′-
untranslated regions and ribosome-binding sequences under the two growth conditions. Our findings provide
potential strategies to optimize the metabolism of syngas fermenting acetogenic bacteria for better productivity.
** This work was supported by the Intelligent Synthetic Biology Center of Global Frontier Project 2011-
0031957 and C1 Gas Refinery Program 2018M3D3A1A01055733 of the National Research Foundation of
Korea, funded by the Ministry of Science, ICT and Future Planning.

Keywords : Acetogenesis, Acetogen, Translational regulation, Wood-Ljungdahl pathway

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6204 Improving CO2 Tolerance by Expression of a Single H+ Pump Enables Microalgae-based

Industrial Flue Gas Valorization

Hong Il CHOI1, Sung-Won HWANG2, Sang Jun SIM1

1
Department of Chemical and Biological Engineering, Korea University, Seoul, Korea, 2Department of
Chemical Engineering, University of Michigan, MI, USA
Corresponding Author Email : [email protected]

Microalgae have garnered much attention as a promising biological platform for CO2 reduction for their ability
to photosynthetically produce various carbon-neutral products using CO2 as a sole carbon source. However, the
real-world application has been severely limited since they are ironically susceptible to elevated CO2. In order to
address this issue, we scrutinized the transcriptome of a CO2 intolerant model microalga, Chlamydomonas
reinhardtii, in response to extremely high CO2 environments to pinpoint the genetic cause and found that
aberrantly low expression of plasma membrane H+-ATPases (PMAs), would be a primary reason for the
susceptibility. Subsequently, we constitutively overexpressed a universally expressible PMA protein in C.
reinhardtii. The single gene overexpression yielded a microalgal strain that exhibit 3.2-fold increased
autotrophic production against high CO2 milieus. The reproducibility of the improvement was confirmed by
proof-of-concept outdoor cultivation with high CO2-containing flue gas emitted from a power plant. The results
suggest that flue gas streams can be valorized by the microalgae when conjugated with this genetic engineering
strategy.

Keywords : Microalgae, CO2 utilization, CO2 tolerance

References
1. A. Solovchenko and I. Khozin-Goldberg, Biotechnol. Lett. 35, 1745 (2013).
2. L. Wei et al., Metab. Eng. 54, 95 (2019).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6205 The Entner–doudoroff Pathway (EDP) Intermediate-inducible Expression System for 2,3-
butanediol Production in Escherichia coli

SATHESH-PRABU CHANDRAN, Sung Kuk LEE

School of Energy and Chemical Engineering, Ulsan National Institute of Science and Technology (UNIST),
Ulsan, Korea
Corresponding Author Email : [email protected]

Recent biotechnological progress, particularly in the areas of genetic/genomic engineering, synthetic biology,
systems biology, and metabolic engineering, offers novel tools to customize cell factories for bio-based
production. The production of bio-based bulk chemicals from renewable bioresources has gained great attention
in the scientific community as the process could avoid and/or minimize the negative impacts of the conventional
chemical-based productions on the environment and depleting resources of natural petrochemicals. An inducible
promoter is a crucial component for microbial-based chemical production. The induction of promoter expression
by the addition of chemical inducers such IPTG is considered the most efficient method. However, it has the
following limitations, such as the requirement of cell growth monitoring for the addition of chemical inducers at
the optimal cell density, not feasible with industrial scale-up, and undesirable because of its toxicity and cost.
Moreover, constitutive expression or strong expression of synthetic pathway genes can cause growth inhibition,
less glucose consumption, and reduce product yield. Thus, inexpensive carbon source- or substrate-inducible
systems are considered as one of the efficient systems for biosynthesis of chemicals, especially at industrial
scale. 2,3-Butanediol (BDO), an important platform chemical, has many industrial applications and is used as a
solvent, a high-grade aviation fuel because of its high-octane number, and a precursor of many synthetic
polymers and resins. In addition, BDO is used in the manufacture of perfumes, fumigants, printing inks,
moistening and softening agents, antifreeze, lubricants, fuel additives, explosives, plasticizes, and pharmaceu-
tical carriers. In the present study, a glucose-inducible gene expression system has been developed using HexR-
Pzwf1 of Pseudomonas putida to induce the metabolic pathways. Since the system is controlled by an Entner–
Doudoroff pathway (EDP) intermediate, the EDP of Escherichia coli was activated by deleting pfkA and gntR
genes. Growth experiment with GFP as a reporter indicated that the induction of this system was tightly
controlled over a wide range of glucose in E. coli without adding any inducer. 2,3-butanediol (BDO) synthetic
pathway genes were expressed by this system in the pfkA-gntR-deleted strain. The resultant engineered strain
harbouring this system efficiently produced BDO with a 71% increased titer than the control strain. The strain
was also able to produce BDO from a mixture of glucose and xylose which is comparable to glucose alone.
Further, the strain produced 11 g/L of BDO at a yield of 0.48 g/g from the hydrolysate of empty palm fruit
bunches. This system can also be applied in many other bio-production processes from lignocellulosic biomass.

Keywords : 2,3-Butanediol (BDO), Entner-Doudoroff pathway, Pseudomonas putida, HexR

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6206 Genome Mining and Reassessment of Streptomyces venezuelae Strains Harboring Novel
Secondary Metabolites Biosynthetic Gene Clusters

Namil LEE1,2, Mira CHOI3, Woori KIM1,2, Suhyung CHO1,2, Bernhard PALSSON4,5,6, Kyoung-Soon JANG3,7,
Byung-Kwan CHO1,2
1
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, Korea,
2
Innovative Biomaterials Research Center, KI for the BioCentury, Korea Advanced Institute of Science and
Technology, Daejeon, Korea, 3Bio-Chemical Analysis Team, Korea Basic Science Institute, Cheongju, Korea,
4
Department of Bioengineering, University of California San Diego, La Jolla, CA, USA, 5Department of
Pediatrics, University of California San Diego, La Jolla, CA, USA, 6Novo Nordisk Foundation Center for
Biosustainability, Technical University of Denmark, Lyngby, Denmark, 7Division of Bio-Analytical Science,
University of Science and Technology, Daejeon, Korea
Corresponding Author Email : [email protected]

Streptomyces strains have been traditionally classified based on morphological and physiological observations.
Therefore, considerable inaccuracies have accumulated in Streptomyces taxonomy, causing the misclassification
of a large number of Streptomyces strains. Here, we present the highly resolved classification of 10
Streptomyces venezuelae strains, using 16S rRNA sequencing, MALDI-TOF MS protein profiling, and whole-
genome mining. The results revealed that only three (ATCC 10595, 21113, and 10712) of the 10 strains could
be classified as S. venezuelae species. Accurate classification, following secondary metabolite biosynthetic gene
cluster mining and targeted LC-MS/MS-based metabolite screening, enabled to identify of the production of 126
secondary metabolites from the 10 S. venezuelae strains. In particular, the comparison of biosynthetic gene
clusters for pyrrolamide-type antibiotics of four S. venezuelae strains revealed that the novel gene, athv28, is
critical in the synthesis of the anthelvencin precursor, 5-amino-3,4-dihydro-2H-pyrrol-2-carboxylate. Our
findings illustrate the importance of the accurate and reliable classification of Streptomyces and better utilization
of misclassified Streptomyces strains for the discovery of novel smBGCs.
** This work was supported by the Bio & Medical Technology Development Program (grant no. 2018M
3A9F3079664 to B.-K.C.) through the National Research Foundation (NRF) funded by the Ministry of Science
and ICT (MSIT). This work was also supported by a grant from the Novo Nordisk Foundation (grant no.
NNF10CC1016517) and KBSI grant (C030440).

Keywords : Streptomyces, classification, genomics, secondary metabolite

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6207 Pig Sera-derived Anti-SARS-CoV-2 Antibodies with Controlled Specificity for Competitive
Immunoassay

Ji-Hong BONG, Jaeyong JUNG, Jeong Soo SUNG, Chang Kyu LEE, Jae-Chul PYUN
Department of Materials Science and Engineering, Yonsei University, Seoul, Korea
Corresponding Author Email : [email protected]

Anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies against nucleoprotein (NP)
were purified from pig sera through two steps: (1) isolation of anti-SARS-CoV-2 NP antibodies using magnetic
beads with immobilized SARS-CoV-2 NP and (2) filtration of anti-SARS-CoV-2 spike protein (SP). The
binding activities of the isolated antibodies were confirmed using immunoassay and immunostaining. The
binding constant (Kd) of the purified anti-SARS-CoV-2 NP antibodies was estimated using a surface plasmon
resonance biosensor (SPR) and calculated to be 185 pM. From the dose-response curve, the limit of detection
was estimated to be 1.02 pM. Finally, a competitive immunoassay was developed for detecting SARS-CoV-2
NP as well as SARS-CoV-2 in the culture fluid using magnetic beads with immobilized anti-SARS-CoV-2 NP
antibodies. The results demonstrated (a) the detection of SARS-CoV-2 in viral fluid and (b) the discrimination
of SARS-CoV-2 from SARS-CoV, MERS-CoV, and CoV strain 229E in viral fluids. Additionally, anti-SARS-
CoV-2 antibodies against SP were obtained when the isolation process was performed using the magnetic beads
with immobilized human SARS-CoV-2 SP.

Keywords : SARS-CoV-2, COVID-19, Antibody, Nucleoprotein, Spike protein, Surface plasmon resonance,
Immunoassay

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

[S6-3]
부문위원회 및 학생세션 III

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6301 Bioproduction of Propionic Acid from Engineered Pseudomonas putida Strain

Rameshwar TIWARI, Sung Kuk LEE

School of Energy and Chemical Engineering, Ulsan National Institute of Science and Technology (UNIST),
Ulsan, Korea
Corresponding Author Email : [email protected]

Propionic acid is an industrially relevant organic acid used in various applications like food, agricultural, and
pharmaceutical industries. However, petrochemical based unsustainable propionic acid production leads to the
development of microbial bioprocess from renewable substrates like lignocellulosic biomass. Levulinic acid is a
γ-keto acid (C5) platform chemical produced by acid-catalyzed dehydration and the hydrolysis of sugars
obtained from lignocellulosic biomass without using expensive hydrolytic enzymes. The metabolic pathway of
levulinic acid utilization was discovered in Pseudomonas putida strain and further used for levulinic acid-based
biorefinery to produce biologically diverse chemicals. Moreover, P. putida is recognized as a safe microbial
host and is emerging as a next-generation industrial workhorse due to its metabolic versatility and remarkable
tolerance to oxidative stress, organic solvents, and aromatic compounds. In present work, levulinic acid
assimilation pathway of P. putida EM42 strain was directed to produce propionic acid by blocking side
pathways and its interconversion into propionyl-CoA. Further, levulinic acid-inducible expression system was
used to express metabolic genes for propionic acid production. Metabolic engineered P. putida strain was used
to optimize the bioprocess for higher production yield of propionic acid. This study revealed the exploitation of
levulinic acid catabolic pathway from P. putida as an alternative route for the sustainable and industrial
production of propionic acid.

Keywords : Propionic acid, Pseudomonas putida, levulinic acid, metabolic engineering

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6302 Metabolic Engineering of Corynebacterium glutamicum for the Enhanced Production of a

Natural Blue Pigment Indigoidine as Sustainable Fabric Dyes

Mohammad Rifqi GHIFFARY1,2, Cindy Pricilia Surya PRABOWO2,3, Komal SHARMA1,2, Sang Yup LEE2,3,4,
Hyun Uk KIM1,2,4
1
Systems Biology and Medicine Laboratory, Department of Chemical and Biomolecular Engineering, Korea
Advanced Institute of Science and Technology (KAIST), Daejeon, Korea, 2Systems Metabolic Engineering and
Systems Healthcare Cross-Generation Collaborative Laboratory, KAIST, Daejeon, Korea, 3Metabolic and
Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engi-
neering, KAIST Institute for BioCentury, KAIST, Daejeon, Korea, 4KAIST Institute for Artificial Intelligence,
BioProcess Engineering Research Center and BioInformatics Research Center, KAIST, Daejeon, Korea
Corresponding Author Email : [email protected]

The industrial production of dyes for textiles mainly involves numerous toxic chemicals which causes severe
water pollution. For this, indigoidine has attracted attention as an alternative natural blue dye, but it is necessary
to achieve a high-level production to compete with existing synthetic blue dyes. Here, we report a metabolically
engineered Corynebacterium glutamicum capable of producing indigoidine to a high concentration with high
productivity. First, blue-pigment indigoidine synthetase (bpsA) gene from Streptomyces lavendulae was
expressed in C. glutamicum, which carries strong fluxes toward L-glutamate, a precursor of indigoidine.
Production performance of this base strain, already producing 7.3 ± 0.3 g/L indigoidine from flask, was further
improved by streamlining intracellular supply of the precursors L-glutamate and L-glutamine, strengthening
phosphotransferase system-independent glucose uptake system, channeling carbon fluxes from glycolysis to
tricarboxylic acid (TCA) cycle, and minimizing byproducts formation. The final strain named BIRU11 produced
49.30 g/L indigoidine with the productivity of 0.96 g/L/h from fed-batch fermentation, the highest titer and
productivity to date. Finally, indigoidine from the fed-batch fermentation of the BIRU11 strain was used to dye
white cotton fabrics to examine its color and performance. This study demonstrates the potential of producing
fabric dyes in a sustainable manner by using a metabolically engineered bacterium.

Keywords : Corynebacterium glutamicum, fabric dye, indigoidine, metabolic engineering, natural blue pigment,
nonribosomal peptide

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6303 Bone Substitute Biomineral Complex Co-derived from Marine Biocalcification and
Biosilicification

Jinyoung YUN1, Yeonsu JEONG1, Onyou NAM2, Ki Baek YEO3, Yun Kee JO4, Hye Ryoung HEO1, Chang Sup
KIM5, Kye Il JOO1, Seung Pil PAC3, EonSeon JIN2, Hyung Joon CHA1
1
Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Korea,
2
Department of Life Science, Hanyang University, Seoul, Korea, 3Department of Biotechnology and Bioinfor-
matics, Korea University, Sejong, Korea, 4Department of Biomedical Convergence Science and Technology,
Kyungpook National University, Daegu, Korea, 5School of Chemistry and Biochemistry, Yeungnam University,
Gyeongsan, Korea
Corresponding Author Email : [email protected]

With aging population, the need for the use of bone graft is increasing to promote healing process of damaged
bone [1]. Bone substitute materials have been mainly developed based on inorganic materials, including calcium
phosphate. However, these graft materials usually act as osteoconductive rather than osteoinductive scaffolds
[2]. To improve bone reconstruction, the combination of several materials have been proposed. However, there
are still no alternatives that can completely replace the existing animal-derived bone graft materials [3]. In this
work, a marine-inspired biomineral complex was suggested as a novel potential bone graft material. The
proposed biosilicified coccolithophore-derived coccoliths using bioengineered mussel adhesive protein show
osteopromotive ability through the synergistic effects of osteoconductivity from calcium carbonate and
osteoinductivity from silica. Its feasibility as a bone graft material was determined by evaluating the in vitro
osteogenic behaviors of preosteoblast cells and in vivo bone regeneration in rat calvarial defect model.
Therefore, the marine-inspired biomineral complex developed in this study could be successfully used in the
field of bone regenerative medicine.

Keywords : coccoliths, biosilica, biomineral, bone substitutes, bone regeneration

References
1. C. J. Damien and J. R. Parsons, J. Appl. Biomater., 2, 187 (1991); W. H. Hiatt et al., J. Periodontology, 49,
495 (1978).
2. A. Brydone, D. Meek, S. Maclaine, Proc. Inst. Mech. Eng., Part H, 224, 1329 (2010).
3. Y. Khan et al, J. Bone Jt. Surg, 90, 36 (2008).

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6304 Amine-catechol Pair Effect on Underwater Adhesion of Marine Mussel

Mincheol SHIN1, Ji Yeon SHIN2, Kyeounghak KIM1, Byeongseon YANG1, Yeonju PARK3, Sila JIN4, Young
Mee JUNG3,4, Jeong Woo HAN1, Nak-Kyoon KIM2, Hyung Joon CHA1
1
Department of chemical engineering, POSTECH, Pohang, Korea, 2Advanced analysis center, KIST, Seoul,
Korea, 3Kangwon radiation convergence research support center, Kangwon National University, Chuncheon,
Korea, 4Department of chemistry, Kangwon National University, Chuncheon, Korea
Corresponding Author Email : [email protected]

Non-covalent interactions have attained many attentions in various areas including molecular recognition,
biological self-assembly, and molecular adhesion. Among them, cation-π interaction has been suggested as
crucial intermolecular interaction in underwater adhesion recently. In specific, several studied have shown that
cation-π interaction is the main contributor to the liquid-liquid phase separation (LLPS) in underwater for
mussel-inspired materials. Also, molecular cohesion forces found out to be tuned by changing π candidate in
underwater. In this study, we focused on how the position of cation sources could affect intermolecular cation-π
interactions in underwater using surface forces apparatus (SFA), Raman spectroscopy, and NMR. Model
peptides were synthesized for nanomechanical studies. Underwater cation-pi interactions were measured by (1)
different aromatic π sources, (2) salt concentrations, and (3) reversibility of non-covalent interactions. This
study could suggest that cation sources might deteriorate intermolecular cation-π interactions and could give
insight on designing biomaterials using in underwater.

Keywords : Underwater adhesion, Mussel adhesion, Dopa, Amine-catechol synergy, Cation-pi interaction,
Noncovalent interaction

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6305 Redox Regulation of Dopa in Interfacial Underwater Adhesion of Marine Mussel

Tae Hee YOON, Mincheol SHIN, Byeongseon YANG, Hyung Joon CHA
Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Korea
Corresponding Author Email : [email protected]

Underwater adhesion has got attention due to its versatility in biomedical applications. Since many biomedical
applications face with a wet environment, underwater adhesion should be applied for adequate contact with
watery surfaces such as tissues. Researchers has focused on marine organisms such as mussel which facilitates
wet adhesion in daily life. Many studies revealed that 3,4-dihydroxyphenylalanine (Dopa) is a key molecule in
underwater adhesion. Dopa can participate in both surface adhesion and cohesion according to its oxidative
states. However, chemical state of Dopa and their role in adhesion is hard to control because it is easily oxidized
in physiological environment and has complicated oxidative pathway. Therefore, to make successful underwater
glue using Dopa, understanding how marine mussel regulates the redox of Dopa in the underwater adhesive
system is essential. In this study, we investigated effect of redox protein on interfacial adhesive protein in
mussel adhesion.

Keywords : Underwater adhesion, 3,4-dihydroxyphenylalanine, Redox regulation

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6306 Electrospun Surgical Dressing with Natural Proteins for Dual Function as Topical Hemostatic
Agent and Anti-adhesion Barrier

Jaeyun LEE1, Eun Jin KIM2, Ki Joo KIM2, Jong Won RHIE2, Kye Il JOO1,3, Hyung Joon CHA1
1
Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, Korea,
2
Department of Plastic and Reconstructive Surgery, The Catholic University of Korea, Seoul, Korea, 3Division
of Chemical Engineering and Materials Science, Ewha Womans University, Seoul, Korea
Corresponding Author Email : [email protected]

Visceral surgery is accompanied by a risk of severe hemorrhage that cannot be controlled by patients’ natural
coagulation ability. Furthermore, the coagulopathies from chronic bleeding or anticoagulant drugs may fall into
acidosis and hypothermia which leading them to fatal trauma triad at emergent state. In addition, wounds with
excessive bleeding may also cause undesired adhesion with surrounding organs, inflammation, infection, and
neuralgia, worsening the surgical prognosis. Therefore, it is necessary for surgical dressing to halt bleeding on
the side contact with wound and prevent adhesion on the outer side. However, most commercial products do not
have both features as topical hemostatic agents and anti-adhesion barriers. There are recent approaches
introducing hydrophobic paraffin, silica, synthetic polymer or carbon nanofiber to existing hemostatic materials
for minimizing adhesion and infection, but these materials are limited to skin wounds due to their low
biocompatibility. In this study, novel dressing for internal use with both blood coagulation and anti-adhesion
function was developed by electrospinning silk fibroin and mussel adhesive protein (MAP). Fibroin has feasible
mechanical property as wound dressing, and its hydrophobicity can be controlled through additional solvent
treatment after electrospinning. DOPA residues present in bioengineered MAP have underwater adhesion ability
to aggregate blood plasma components and activate platelets. The functionality, biocompatibility, and
biodegradability of the both proteins in the nanofiber were verified through in vitro and in vivo experiments. In
anticoagulated animal tests, the nanofibrous membrane promoted blood clotting and protected wound areas,
confirming the feasibility as a novel surgical dressing.

Keywords : silk fibroin, mussel adhesive protein, topical hemostatic agent, anti-adhesion barrier, electrospun
nanofiber

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2021 한국생물공학회 춘계학술발표대회 및 국제심포지엄 Emerging Trends in Biotechnology after the Pandemic
2021 KSBB Spring Meeting and International Symposium

S6307 A Simple and Efficient PCR-based Genome Walking Technique to Identify Transgene
Integration Sites in CHO Cells

Hye-Jin HAN, Jong Youn BAIK

Department of Biological Sciences and Bioengineering, Inha University, Incheon, Korea
Corresponding Author Email : [email protected]

Chinese hamster ovary (CHO) cell lines have usually been developed by random integration, yielding
heterogeneous clones with various expression level and stability characteristics due to the transgene insertion
sites in genome. So, it is worth analyzing them for better characterization of cell lines [1]. While next-generation
sequencing is commonly used to locate the insertion site, it requires complicated process and analysis of
sequence reads. Moreover, it is a time-consuming (4-6 weeks) and costly ($2,000-3,000) method. Hence, we
adapted splinkerette-PCR to identify the transgene insertion site with less resources (both time and cost). As a
proof-of-concept experiment, a plasmid vector was cleaved using restriction enzymes, ligated with splinkerette
adaptors that inhibit adaptor-adaptor amplificon [2], PCR-amplified, and run on a DNA gel to visualize the PCR
product with an expected size. After the selection of restriction enzymes that have the highest cutting
frequencies using a bioinformatics tool, this technique was then applied to the CHO genome to analyze the
integration sites of a recombinant protein gene. In conclusion, this method would help to provide the genomic
information with more affordable options in the process of cell line development.

Keywords : Chinese hamster ovary (CHO) cells, Integration site, Unknown DNA sequence, Splinkerette-PCR
References
1. L. M. Grav, D. Sergeeva, J. S. Lee, I. M. D. Mas, N. E. Lewis, M. R. Andersen, L. K. Nielsen, G. M. Lee
and H. F. Kildegaard, ACS Synth. Biol. 7, 2148-2159 (2018).
2. A. G. Uren, H. Mikkers, J. Kool, L. V. D. Weyden, A. H. Lund, G. H. Wilson, R. Rance, J. Jonkers, M. V.
Lohuizen, A. Berns and D. J. Adams, Nat. Protoc. 4(5), 789-798 (2009).

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2021 KSBB Spring Meeting and International Symposium

S6308 Bilirubin-encapsulated Albumin Nanoparticles for Cancer Treatment; Synthesis,

Characterization, and in-vitro Cellular Assay

Seyedmohammad MOOSAVIZADEH1, Gyu Hyeok LEE2, Dae Hee LEE2, Sung In LIM1
1
Department of Chemical Engineering, Pukyong National University, Busan, Korea, 2Department of Marine
Food Science and Technology, Gangneung-Wonju National University, Gangneung, Korea
Corresponding Author Email : [email protected]

Bilirubin (BR) is the end-product of heme catabolism which occurs abundantly in the blood plasma. Bilirubin is
an endogenous antioxidant capable of scavenging different reactive oxygen species (ROS), thereby playing a
role in protecting cells and tissues from oxidative damage. Indeed, numerous experimental studies proved the
potential of bilirubin for anti-inflammatory and anti-cancer activities. However, its low water-solubility and high
sensitivity to oxygen restrict further clinical developments, requiring an appropriate drug carrier capable of
stably delivering a defined dosage to the site of action. Albumin is the most abundant protein in blood with a
long circulatory half-life, with the capability to bind nutrients, metabolites, and metals, lending itself to a
promising drug carrier. Moreover, albumin is specifically localized to tumors or inflamed tissues in either an
active or a passive manner. Recently, serum albumin nanoparticles, i.e., BSA-NPs, have been used for
delivering various types of therapeutic agents, encapsulated inside or modified on the surface. In this study, we
explored how to synthesize and characterize albumin nanoparticles containing hydrophobic bilirubin by the
Desolvation method and measure its toxicity on normal and cancerous cells. Using this method, albumin
nanoparticles encapsulated bilirubin with a stable size around 120 nm, and a spherical morphology were
obtained. The entrapment efficiency (EE) was around 60%, and the drug loading (DL) was around 10%. The in
vitro cell assay of nanoparticles on normal and cancerous cells showed low toxicity on normal cells, under 10%,
in contrast to more than 60% toxicity on cancerous cells.

Keywords : Albumin nanoparticle, drug delivery, cancer therapy

References
1. C. Weber, et al. Int. J. Pharm. 194, 91-102 (2000).
2. A. Jahanban-Esfahlan, et al. Int. J. Biol. Macromol. 91, 703-709 (2016).
3. Lee Y, et al. Angew Chemie-Int. Ed. 55(36):10676-10680 (2016).

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2021 KSBB Spring Meeting and International Symposium

S6309 Assessing the Role of Cellular dsRNAs in the Pathogenesis of Sjogren's Syndrome and as the
Therapeutic Target in Autoimmunity

Jimin YOON1, Min-Seok LEE1, Ahsan Ausaf ALI1, Ye Rim OH2, Yong Seok CHOI2, Namseok LEE1, Seunghee
CHA3, Joon Young HYON4, Yun Jong LEE5,6, Sung Gap IM1,7, Yoosik KIM1,8
1
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology
(KAIST), Daejeon, Korea, 2Medical Science Research Institute, Seoul National University Bundang Hospital,
Seongnam, Korea, 3Department of Oral and Maxillofacial Diagnostic Sciences, University of Florida College of
Dentistry, Gainesville, FL, USA, 4Department of Ophthalmology, Seoul National University Bundang Hospital,
Seongnam, Korea, 5Division of Rheumatology, Department of Internal Medicine, Seoul National University
Bundang Hospital, Seongnam, Korea, 6Department of Internal Medicine, Department of Internal Medicine,
Seoul National University College of Medicine, Seoul, Korea, 7KI for NanoCentury (KINC), KAIST, Daejeon,
Korea, 8KI for Health Science and Technology (KIHIST), KAIST, Daejeon, Korea
Corresponding Author Email : [email protected]

Sjӧgren’s syndrome (SS) is a systemic autoimmune disease that mainly targets the exocrine glands, resulting in
impaired saliva and tear secretion. To date, SS is under-diagnosed due to complex immunopathogenesis and the
lack of biomarkers. Here, we investigated the role of cellular double-stranded RNAs, especially originated from
the mitochondria (mt-dsRNAs) in pathogenesis of SS. We found that mt-dsRNAs were elevated in saliva/tears
of SS patients and were related to exocrine dysfunction and glandular lymphocytic infiltration. Using our unique
3D culture system, we showed that dsRNA stress led to increased mt-dsRNAs and their cytosolic export,
accompanied by characteristic molecular phenotypes of SS. Moreover, mt-dsRNA induction occurred
downstream of the JAK1 pathway. Interestingly, suppression of mtRNA induction by antioxidant, JAK1
inhibitor, or mtRNA polymerase inhibitor reverse