Biodiesel Production Using Bacterial Fatty Acids Extracts

Elsevier, 2020

Increasing population load and an altered lifestyle attitude are exerting extra pressure on the production market, to satisfy the demands and desire of society. The recently developed production and consumption models largely rely on fossil-based resources, which are affecting the environment and natural resources adversely. The cost-effective production of biological materials is an emerging sector with remarkable future prospects and provides many business opportunities. With time the research endeavors are gradually shifting toward bacterial lipids-derived biofuel production, which is more suitable and compatible for an industrial application. The major challenge in the overall process of the production of lipids-derived fuels from microbes is the involved carbon source as it contributes to more than half of the production cost. Therefore the production of lipids and biodiesel from bacteria using different waste materials as carbon source involving the application of advanced biotechnological tools, and modified transesterification reactions will make the biodiesel production cost effective.

Rasayan Journal of Chemistry

The exploration of lipolytic bacteria presents its challenges in being able to supply lipase as a catalyst in the production of biodiesel. The compost is chosen to get the bacteria because of the lipid and fatty acids component contained in it. The research was developed to isolate the lipase-producing bacteria and determine the enzyme ability in bioconversion of lipid to biodiesel. The bacterial screening was conducted in Luria Bertani media with additional olive oil 1% (v/v) as an inducer. The colonies surrounded by bright areas were separated from others because they indicated having lipolytic activity. Identification of selected isolate was carried out according to the 16srRNA fragment DNA analysis. The fragment showed a score of nucleotide identity as 98% with the Proteus sp. The isolated bacteria grow well in LB media forming an exponential phase from 2 to 11 h, and have a double-time of cell growth at the 4 h. It produced lipase optimally at the 19 hours with its activity of 1.771 U/mL. The enzyme showed optimum activity at 45 o C and pH 8. The lipase showed high activity in t-butanol then followed by sequentially in a mixed solvent (methanol and tbutanol), and a single solvent of isopropanol, methanol, and n-hexane. It exhibited good performance for the biodiesel production from microalgal oil, yielded a total fatty acid methyl esters (FAME) as 2.75% (v/v). The fatty acid composition of the methyl esters was composed of 9 octadecanoic acid and hexadecanoic acid.

Brazilian Archives of Biology and Technology, 2012

Since centuries vegetable oils are consumed as human food but it also finds applications in biodiesel production which is attracting more attention. But due to being in competition with food it could not be sustainable and leads the need to search for alternative. Nowdays microbes-derived oils (single cell oils) seem to be alternatives for biodiesel production due to their similar composition to that of vegetable oils. However, the cold flow properties of the biodiesel produced from microbial oils are unacceptable and have to be modified by an efficient transesterification. Glycerol which is by product of transesterification can be valorised into some more useful products so that it can also be utilised along with biodiesel to simplify the downstream processing. The review paper discusses about various potent microorganisms for biodiesel production, enzymes involved in the lipid accumulation, lipid quantification methods, catalysts used in transesterification (including enzymatic catalyst) and valorisation of glycerol.

Bioresource Technology Reports

A B S T R A C T Rising level of CO 2 , increasing demands and price of fossil fuel, generation of waste are the major challenges of the modern society. Biorefinery approach for production of biodiesel using waste as feedstock could be an alternative of these problems. The present research demonstrates the use of municipal secondary sludge as growth media for production of biodiesel by chemolithotrophic, oleaginous bacteria Serratia sp. ISTD04. The characterization of bacteria and biodiesel was performed by TEM and GC–MS, NMR respectively. The hydrocarbon range, C-7 to C-25 and suitable saturated and unsaturated fatty acid composition, makes it an impending fuel. Optimization of in-situ transesterification process parameters increased the yield by 25% and similarly increases in transesterification efficiency from 74% to 94%. The fuel properties (Total acid value, Cetane number, Cloud point, Pour point, Heating value, Kinematic viscosity) of 5% blended biodiesel was analyzed by ASTM standard methods and its shows good fuel behavior.

Journal of Bioscience and Bioengineering, 2018

Biodiesel is one of the best alternative to depleting fossil fuels for transport sector. However, biodiesel production via lipase mediated transesterification has limitation of high costing microbial enzymes. In order to overcome this limitation, a process of sequential treatment of oil industry wastewater using isolated lipolytic bacterial strains and biodiesel production from non-edible plant oils was studied. In this study, efficient lipase producing bacteria were isolated and evaluated for production of biodiesel from mustard, soybean, jatropha and taramira oils utilizing methanol for the transesterification of oils and bioremediation. Selected strains were then identified, using 16s rRNA sequencing. Further, Bacillus subtilis strain Q1 KX712301 was optimized for biodiesel production from non-edible taramira oil via Plackett-Burman and central composite design. Highest volumetric yield of biodiesel obtained was 102% at optimized parameters. Finally, a sequential bioremediation of vegetable oil contaminated wastewater and then microbial production of biodiesel from non-edible taramira oil was carried out using efficient lipase producer B. subtilis strain Q1 at optimized conditions. During sequential process, complete chemical oxigen demand reduction of oil containing wastewater and theoretical volumetric yield of biodiesel was achieved. Gas chromatography/mass spectrometry chromatogram revealed that the total fatty acid methyl ester content of the produced biodiesel was >98% which is in accordance with the biodiesel quality standards specified by both ASTM and EU-14103.

Biodiesel, an alternative to petroleum oil has gained significant attention from the research community because of its high energy content and good compatibility with existing engine systems. It can be produced from many different sources, such as animals, plants, and microbes. In this study, we demonstrated the overproduction of fatty acid methyl esters (FAMEs) using a synthetic consortium of manA mutant Streptomyces coelicolor with Ralstonia eutropha. The synthetic consortium of S. coelicolorDmanA: R. eutropha produced 114 mg/L fatty acids, which is 124% higher than the amount produced using S. coelicolor alone. Overall, the fatty acids produced by the consortia S. coelicolorDmanA: R. eutropha were composed of medium chain fatty acid (MCFA): long chain fatty acid (LCFA): very long chain fatty acid (VLCFA) in 8.75: 91.0: 0.25 proportion, and contained 75% saturated and 25% unsaturated fatty acids, which resulted in FAMEs with better cetane number (65) and oxidation stability (76 h) than the fatty acids produced by one strain alone. Nile red staining and subsequent fluorescence spectroscopy revealed S. coelicolorDmanA as good candidate for triacylglycerol (TAG) accumulation. Phospholipid-derived fatty acids (PLFA) analysis of consortia shows that S. coelicolorDmanA and R. eutropha synergistically support each other’s growth. The results suggest that the synthetic consortium provides an approach for biodiesel production along with improved quality.

INTERNATIONAL CONFERENCE ON ENERGY AND ENVIRONMENT (ICEE 2021), 2021

In recent times, increased consumption of fossil fuels due to overpopulation, increasing energy demand have resulted in global warming and climate changes due to the emission of greenhouse gases from these fuels. To compensate for the requirement of fossil fuel, renewable biofuels especially, biodiesel are preferred for satisfying the energy demand. Since biodiesel from edible feedstocks is deemed against "food vs. fuel" policies, first-generation biofuels are not regarded as suitable and sustainable, inspite of growing energy demand. Following this, Second generation biodiesel production from lingo-cellulosic-based feedstocks is the least recommended because of their meticulous procedures and high capital investments. Considering these setbacks, edible feedstocks have been replaced with microbial feedstocks, which are later on used for extracting oil and then transesterified into biodiesel. Even though biodiesel from microbes have their setbacks, they are widely appreciated due to their merits which include short life span, ability to grow on multiple environments and ability to remediate different polluted environmental conditions. Presently, this paper focuses on summarizing the production of biodiesel from various microbial species.

PLoS ONE, 2014

Fossil fuels are consumed so rapidly that it is expected that the planet resources will be soon exhausted. Therefore, it is imperative to develop alternative and inexpensive new technologies to produce sustainable fuels, for example biodiesel. In addition to hydrolytic and esterification reactions, lipases are capable of performing transesterification reactions useful for the production of biodiesel. However selection of the lipases capable of performing transesterification reactions is not easy and consequently very few biodiesel producing lipases are currently available. In this work we first isolated 1,016 lipolytic microorganisms by a qualitative plate assay. In a second step, lipolytic bacteria were analyzed using a colorimetric assay to detect the transesterification activity. Thirty of the initial lipolytic strains were selected for further characterization. Phylogenetic analysis revealed that 23 of the bacterial isolates were Gram negative and 7 were Gram positive, belonging to different clades. Biofuel production was analyzed and quantified by gas chromatography and revealed that 5 of the isolates produced biofuel with yields higher than 80% at benchtop scale. Chemical and viscosity analysis of the produced biofuel revealed that it differed from biodiesel. This bacterial-derived biofuel does not require any further downstream processing and it can be used directly in engines. The freeze-dried bacterial culture supernatants could be used at least five times for biofuel production without diminishing their activity. Therefore, these 5 isolates represent excellent candidates for testing biofuel production at industrial scale.

Bioprocess and Biosystems Engineering, 2012

In an attempt to isolate a biocatalyst able to catalyze biodiesel production from microbial source, Streptomyces sp. CS326 was screened from hundreds of soil isolates collected from various parts of Korea. In 16S rRNA sequence analysis, the strain showed high degree of similarity with Streptomyces xanthocidicus (99.79%); therefore, it is classified as Streptomyces sp. CS326. An extracellular lipase produced by the strain (LP326) was purified using a single step gel permeation chromatography on Sepharose CL-6B. Molecular weight of LP326 was estimated to be 17,000 Da by SDS-PAGE. The activity was optimum at 40°C and pH 7.0 and was stable at pH 5.0-8.0 and below 50°C. It preferred p-nitrophenyl palmitate (C16), a long chain substrate; and K m and V max for the substrate were determined to be 0.24 mM and 4.6 mM/min mg, respectively. First 10 N-terminal amino acid sequences were APDLVALQSE, which are different from so far reported lipases. LP326 catalyzed biodiesel production using methanol and various oils; therefore, the enzyme can be applicable in the field of biofuel.