Papers by Felipe Chambergo
World Academy of Science, Engineering and Technology, International Journal of Biotechnology and Bioengineering, 2018
Genomic information provides the first stage in understanding the instructions for the synthesis ... more Genomic information provides the first stage in understanding the instructions for the synthesis of all of life's molecular machines and the systems needed to control and operate them. The interaction among biological, physical and computing sciences has provided the bases for the creation of a great number of bioinformatic tools [1]. Our laboratory uses the multicellular microorganism Trichoderma reesei as a model system to study the genomic regulatory networks for energy production in eukaryotes. During the developing of an "Expressed Sequence Tags" (ESTs) database for T. reesei [2], we achieved a vast amount of information, making it almost impossible to handle all the data obtained without the use of computational tools.
Different approaches have been explored to study skin biology, including the use of stem cells. M... more Different approaches have been explored to study skin biology, including the use of stem cells. Mesenchymal stem cells (MSC) from umbilical cord can be safely and easily obtained, however a simple strategy to monitor their differentiation is essential. Involucrin is a marker of keratinocyte terminal differentiation, and its promoter (pINV) directs stratum-specific expression of this protein. We designed a reporter system containing EGFP under control of pINV to assess MSC differentiation into keratinocytes. The functional sequence of pINV was inserted into a lentiviral vector, originating LeGO-GpINV. MSC were transduced with the LeGO-GpINV and induced to differentiate into keratinocytes upon cultivation with Keratinocyte Serum Free Medium supplemented. MSC differentiation was confirmed by morphological changes and by the expression of epidermal markers, by flow cytometry, quantitative PCR and western blot. The activity of kallikreins 5, 6 and 7 was detected using fluorogenic substra...
Journal of Cultural Heritage, 2015
This study was aimed to describe fungal communities found in biofilms growing on earthen walls (r... more This study was aimed to describe fungal communities found in biofilms growing on earthen walls (rammed earth, wattle-and-daub and adobe) of rural and semi-urban historical buildings representative of the first phase of the Brazilian coffee cycle (1820-1880), in the upper basin of the Paraiba do Sul river, São Paulo State. The relationship between substrates and such surface-associated microbial communities was also investigated. Fungal biodiversity was significantly higher in rammed earth with respect to the other two techniques. Granulometric analysis showed that rammed earth also contained a higher percentage of coarse soil fraction which is likely to favour the accumulation of water and organic matter. Cellulolytic activity tested positive for the majority of fungi and acidification test showed that fungi exhibit elevated acidifying capacity suggesting that biodeterioration may occur through acid metabolites.
Mycological Progress, 2014
Cerrado (Brazilian savanna) areas present a high number of endemic and unknown species and conseq... more Cerrado (Brazilian savanna) areas present a high number of endemic and unknown species and consequently have been included among the world's biodiversity hotspots. After a survey of zygomycetes from Cerrado areas of the "Reserva Biológica de Mogi Guaçu" (RBMG) in São Paulo State, we have found a Mucor-like fungus that produces very large sporangiospores and clusters of giant cells visible to the naked-eye. This isolate was characterized morphologically, based on partial LSU (28S) and complete ITS rDNA sequences. Maximum likelihood phylogenetic analyses support its inclusion within the recently emended genus Backusella.
Journal of Biological Chemistry, 2002
This work is dedicated to Professor Metry Bacila on his 80th birthday.
Anesthesia & Analgesia, 2003
C-fos and Fos expression, frequently used as a neural nociceptive marker, is altered by many drug... more C-fos and Fos expression, frequently used as a neural nociceptive marker, is altered by many drugs. The effects of histamine receptor antagonists on c-fos messenger (m)RNA expression are unknown. We examined the effect of local and systemic administration of pyrilamine (H 1 receptor antagonist) and cimetidine (H 2 receptor antagonist) on the nociceptive flinching behavior elicited by injection of 50 L of 1% formalin into the dorsal region of the hind paw of rats. Nociceptive flinching behavior was observed for 45 min, and the rats were then killed and lumbar spinal cord obtained for c-fos mRNA expression, measured using the Northern blot hybridization technique. Systemic administration of pyrilamine and Materials and Methods All experiments were performed on Wistar male rats (300-350 g) supplied by our own breeding facilities at the University of São Paulo School of Medicine. The rats had free access to food and water. The experiments were performed after the approval of the Bioethics Committee of the Hospital of Clinics of the University of São Paulo School of Medicine and according to the Committee for Research and Ethical
Carbohydrate Research, 2011
The isolation of a dimeric 29.8-kDa lectin (LFL) from dried Lactarius flavidulus fruit bodies is ... more The isolation of a dimeric 29.8-kDa lectin (LFL) from dried Lactarius flavidulus fruit bodies is reported herein. The chromatographic procedure utilized comprised anion-exchange chromatography on DEAE-cellulose, cation-exchange chromatography on CM-cellulose, anion-exchange chromatography on SP-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The hemagglutinating activity of LFL was inhibited by a variety of simple sugars, such as lactose, p-nitrophenyl aD -glucopyranoside, p-nitrophenyl b-D-glucopyranoside and inositol, and by the polysaccharide inulin. The activity of LFL was stable up to 40°C. There was a precipitous drop in activity when the temperature was elevated to 50°C. Hemagglutinating activity was retained when LFL was exposed to 6.25-12.5 mM HCl and NaOH. The activity was potently inhibited by Fe 2+ and Fe 3+ ions, and slightly inhibited by Al 3+ and Mn 2+ ions. LFL suppressed the proliferation of hepatoma (HepG2) and leukemic (L1210) cells with an IC 50 of 8.90 and 6.81 lM, respectively. It inhibited the activity of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) with an IC 50 of 5.68 lM. However, LFL did not exhibit antifungal activity.
International Journal of Biological Macromolecules, 2019
Epoxide hydrolases (EHs) are enzymes involved in the metabolism of endogenous and exogenous epoxi... more Epoxide hydrolases (EHs) are enzymes involved in the metabolism of endogenous and exogenous epoxides, and the development of EH inhibitors has important applications in the medicine. In humans, EH inhibitors are being tested in the treatment of cardiovascular diseases and show potent anti-inflammatory effects. EH inhibitors are also considerate promising molecules against infectious diseases. EHs are functionally very well studied, but only a few members have its three-dimensional structures characterized. Recently, a new EH from the filamentous fungi Trichoderma reseei (TrEH) was reported, and a series of urea or amide-based inhibitors were identified. In this study, we describe the crystallographic structures of TrEH in complex with five different urea or amidebased inhibitors with resolutions ranging from 2.6 to 1.7 Å. The analysis of these structures reveals the molecular basis of the inhibition of these compounds. We could also observe that these inhibitors occupy the whole extension of the active site groove and only a few conformational changes are involved. Understanding the structural basis EH interactions with different inhibitors might substantially contribute for the study of fungal metabolism and in the development of novel and more efficient antifungal drugs against pathogenic Trichoderma species.
Applied and Environmental Microbiology, 2018
The hypersaline Kebrit Deep brine pool in the Red Sea is characterized by high levels of toxic he... more The hypersaline Kebrit Deep brine pool in the Red Sea is characterized by high levels of toxic heavy metals. Here we describe two structurally related mercuric reductases (MerAs) from this site, which were expressed in Escherichia coli. Sequence similarities suggest that both genes are derived from Proteobacteria, most likely β- or γ-Proteobacteria. We show that one of the enzymes (K35NH) is strongly inhibited by NaCl, while the other (K09H) is activated in a NaCl-dependent manner. We infer from this difference that the two forms might support detoxification of mercury in bacterial microorganisms that employ the compatible solutes and salt-in strategies, respectively. Three-dimensional structure modeling shows that all amino-acid substitutions unique to each type are located outside the domain responsible for formation of the active MerA homodimer, and the vast majority of these are found on the surface of the molecule. Moreover, K09H exhibits the predominance of acidic over hydroph...
Molecular Biology Reports, 2018
Epoxide hydrolases (EHs) are present in all living organisms and catalyze the hydrolysis of epoxi... more Epoxide hydrolases (EHs) are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EH are involved in the metabolism of endogenous and exogenous epoxides, and thus have application in pharmacology and biotechnology. In this work, we describe the substrates and inhibitors selectivity of an epoxide hydrolase recently cloned from the filamentous fungus Trichoderma reesei QM9414 (TrEH). We also studied the TrEH urea-based inhibitors effects in the fungal growth. TrEH showed high activity on radioative and fluorescent surrogate and natural substrates, especially epoxides from docosahexaenoic acid. Using a fluorescent surrogate substrate, potent inhibitors of TrEH were identified. Interestingly, one of the best compounds inhibit up to 60% of T. reesei growth, indicating an endogenous role for TrEH. These data make TrEH very attractive for future studies about fungal metabolism of fatty acids and possible development of novel drugs for human diseases.
Biochimica et biophysica acta, Aug 11, 2017
Epoxide hydrolases (EHs) are enzymes that have high biotechnological interest for the fine and tr... more Epoxide hydrolases (EHs) are enzymes that have high biotechnological interest for the fine and transformation industry. Several of these enzymes have enantioselectivity, which allows their application in the separation of enantiomeric mixtures of epoxide substrates. Although two different families of EHs have been described, those that have the α/β-hidrolase fold are the most explored for biotechnological purpose. These enzymes are functionally very well studied, but only few members have three-dimensional structures characterised. Recently, a new EH from the filamentous fungi Trichoderma reseei (TrEH) has been discovered and functionally studied. This enzyme does not have high homology to any other EH structure and have an enatiopreference for (S)-(-) isomers. Herein we described the crystallographic structure of TrEH at 1.7Å resolution, which reveals features of its tertiary structure and active site. TrEH has a similar fold to the other soluble epoxide hydrolases and has the two ...
Data in Brief, 2016
The data presented in this article are related to the research article entitled "Epoxide hydrolas... more The data presented in this article are related to the research article entitled "Epoxide hydrolase of Trichoderma reesei: Biochemical properties and conformational characterization" [1]. Epoxide hydrolases (EHs) are enzymes that catalyze the hydrolysis of epoxides to the corresponding vicinal diols. This article describes the optimal parameters for the colorimetric red assay to determine the enzymatic activity, with an emphasis on the characterization of the kinetic parameters, pH optimum and thermal stability of this enzyme. The effects of reagents that are not resistant to oxidation by sodium periodate on the reactions can generate false positives and interfere with the final results of the red assay.
International journal of biological macromolecules, Jan 10, 2016
Epoxide hydrolases (EHs) are enzymes that are present in all living organisms and catalyze the hy... more Epoxide hydrolases (EHs) are enzymes that are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EHs have biotechnological potential in chiral chemistry. We report the cloning, purification, enzymatic activity, and conformational analysis of the TrEH gene from Trichoderma reesei strain QM9414 using circular dichroism spectroscopy. The EH gene has an open reading frame encoding a protein of 343 amino acid residues, resulting in a molecular mass of 38.2kDa. The enzyme presents an optimum pH of 7.2, and it is highly active at temperatures ranging from 23 to 50°C and thermally inactivated at 70°C (t1/2=7.4min). The Michaelis constants (Km) were 4.6mM for racemic substrate, 21.7mM for (R)-(+)-styrene oxide and 3.0mM for (S)-(-)-styrene oxide. The kcat/Km analysis indicated that TrEH is enantioselective and preferentially hydrolyzes (S)-(-)-styrene oxide. The conformational stability studies suggested that, despite the extreme condi...
Applied Microbiology and Biotechnology, 2016
Fungal habitats include soil, water, and extreme environments. With around 100,000 fungus species... more Fungal habitats include soil, water, and extreme environments. With around 100,000 fungus species already described, it is estimated that 5.1 million fungus species exist on our planet, making fungi one of the largest and most diverse kingdoms of eukaryotes. Fungi show remarkable metabolic features due to a sophisticated genomic network and are important for the production of biotechnological compounds that greatly impact our society in many ways. In this review, we present the current state of knowledge on fungal biodiversity, with special emphasis on filamentous fungi and the most recent discoveries in the field of identification and production of biotechnological compounds. More than 250 fungus species have been studied to produce these biotechnological compounds. This review focuses on three of the branches generally accepted in biotechnological applications, which have been identified by a color code: red, green, and white for pharmaceutical, agricultural, and industrial biotechnology, respectively. We also discuss future prospects for the use of filamentous fungi in biotechnology application.
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Papers by Felipe Chambergo