Name: Abordagens de Biologia Estrutural e Química Analítica para Caracterização de Enzimas Metabólicas de C-Glicosídeos na Microbiota Intestinal Humana (Vídeo) | JoVE Journal
Uploaded: 2025-05-23T12:50:00.000+00:00
Duration: 13 min 35 s
Description: Aprenda a caracterizar enzimas metabólicas de C-glicosídeos da microbiota intestinal humana usando cristalografia de proteínas, LC-MS/MS e RNM. Este método integra biologia estrutural e química analítica.

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5 minutes

<ol><li>Gomaa, E. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Human+gut+microbiota%2Fmicrobiome+in+health+and+diseases%3A+a+review%5BTitle%5D)+AND+%22Antonie+Van+Leeuwenhoek%22%5BJournal%5D)">Human gut microbiota/microbiome in health and diseases: a review.</a> Antonie Van Leeuwenhoek. 113 (12), 2019-2040 (2020).</li>
<li>Thursby, E., Juge, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Introduction+to+the+human+gut+microbiota%5BTitle%5D)+AND+%22Biochem+J%22%5BJournal%5D)">Introduction to the human gut microbiota.</a> Biochem J. 474 (11), 1823-1836 (2017).</li>
<li>Nie, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Gut+symbionts+alleviate+MASH+through+a+secondary+bile+acid+biosynthetic+pathway%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">Gut symbionts alleviate MASH through a secondary bile acid biosynthetic pathway.</a> Cell. 187 (11), 2717-2734.e33 (2024).</li>
<li>Xie, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Comparison+of+flavonoid+O-glycoside%2C+C-glycoside+and+their+aglycones+on+antioxidant+capacity+and+metabolism+during+in+vitro+digestion+and+in+vivo%5BTitle%5D)+AND+%22Foods%22%5BJournal%5D)">Comparison of flavonoid O-glycoside, C-glycoside and their aglycones on antioxidant capacity and metabolism during in vitro digestion and in vivo.</a> Foods. 11 (6), (2022).</li>
<li>Zhang, S. -J., Guo, J. -R., Kang, A., Di, L. -Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Advance+in+studies+on+gut+microbiota+in+de-glycosylation+of+traditional+Chinese+medicine+glycosides%5BTitle%5D)+AND+%22Zhongguo+Zhong+yao+za+zhi%3D+Zhongguo+Zhongyao+ZazhiChina+Journal+of+Chinese+Materia+Medica%22%5BJournal%5D)">Advance in studies on gut microbiota in de-glycosylation of traditional Chinese medicine glycosides.</a> Zhongguo Zhong yao za zhi= Zhongguo Zhongyao ZazhiChina Journal of Chinese Materia Medica. 38 (10), 1459-1466 (2013).</li>
<li>Xie, G. -Z., Hui, H. -Y., Peng, M. -J., Tan, Z. -J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Biotransformation+of+glycosides+in+herbal+medicine+by+gut+microbiota%5BTitle%5D)+AND+%22World+Chinese+Journal+of+Digestology%22%5BJournal%5D)">Biotransformation of glycosides in herbal medicine by gut microbiota.</a> World Chinese Journal of Digestology. 26 (4), 221(2018).</li>
<li>Vuong, T. V., Wilson, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Glycoside+hydrolases%3A+catalytic+base%2Fnucleophile+diversity%5BTitle%5D)+AND+%22Biotechnol+Bioeng%22%5BJournal%5D)">Glycoside hydrolases: catalytic base/nucleophile diversity.</a> Biotechnol Bioeng. 107 (2), 195-205 (2010).</li>
<li>Davies, G., Henrissat, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Structures+and+mechanisms+of+glycosyl+hydrolases%5BTitle%5D)+AND+%22Structure%22%5BJournal%5D)">Structures and mechanisms of glycosyl hydrolases.</a> Structure. 3 (9), 853-859 (1995).</li>
<li>Ito, T., Fujimoto, S., Shimosaka, M., Taguchi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Production+of+C-glucosides+of+flavonoids+and+related+compounds+by+Escherichia+coli+expressing+buckwheat+C-glucosyltransferase%5BTitle%5D)+AND+%22Plant+Biotechnology%22%5BJournal%5D)">Production of C-glucosides of flavonoids and related compounds by Escherichia coli expressing buckwheat C-glucosyltransferase.</a> Plant Biotechnology. 31 (5), 519-524 (2014).</li>
<li>Wei, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Discovery+and+mechanism+of+intestinal+bacteria+in+enzymatic+cleavage+of+C-C+glycosidic+bonds%5BTitle%5D)+AND+%22Appl+Microbiol+Biotechnol%22%5BJournal%5D)">Discovery and mechanism of intestinal bacteria in enzymatic cleavage of C-C glycosidic bonds.</a> Appl Microbiol Biotechnol. 104, 1883-1890 (2020).</li>
<li>Che, Q. M., Akao, T., Hattori, M., Kobashi, K., Namba, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Metabolism+of+aloesin+and+related+compounds+by+human+intestinal+bacteria%3A+a+bacterial+cleavage+of+the+C-glucosyl+bond+and+the+subsequent+reduction+of+the+acetonyl+side+chain%5BTitle%5D)+AND+%22Chem+Pharm+Bull+%28Tokyo%29%22%5BJournal%5D)">Metabolism of aloesin and related compounds by human intestinal bacteria: a bacterial cleavage of the C-glucosyl bond and the subsequent reduction of the acetonyl side chain.</a> Chem Pharm Bull (Tokyo). 39 (3), 704-708 (1991).</li>
<li>Wei, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Discovery+and+mechanism+of+intestinal+bacteria+in+enzymatic+cleavage+of+C-C+glycosidic+bonds%5BTitle%5D)+AND+%22Appl+Microbiol+Biotechnol%22%5BJournal%5D)">Discovery and mechanism of intestinal bacteria in enzymatic cleavage of C-C glycosidic bonds.</a> Appl Microbiol Biotechnol. 104 (5), 1883-1890 (2020).</li>
<li>HATTORI, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Metabolism+of+barbaloin+by+intestinal+bacteria%5BTitle%5D)+AND+%22Chem+Pharm+Bull+%28Tokyo%29%22%5BJournal%5D)">Metabolism of barbaloin by intestinal bacteria.</a> Chem Pharm Bull (Tokyo). 36 (11), 4462-4466 (1988).</li>
<li>Che, Q. -M., Akao, T., Hattori, M., Kobashi, K., Namba, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Isolation+of+a+human+intestinal+bacterium+capable+of+transforming+barbaloin+to+aloe-emodin+anthrone1%5BTitle%5D)+AND+%22Planta+medica%22%5BJournal%5D)">Isolation of a human intestinal bacterium capable of transforming barbaloin to aloe-emodin anthrone1.</a> Planta medica. 57 (01), 15-19 (1991).</li>
<li>Braune, A., Blaut, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Deglycosylation+of+puerarin+and+other+aromatic+C-glucosides+by+a+newly+isolated+human+intestinal+bacterium%5BTitle%5D)+AND+%22Environ+Microbiol%22%5BJournal%5D)">Deglycosylation of puerarin and other aromatic C-glucosides by a newly isolated human intestinal bacterium.</a> Environ Microbiol. 13 (2), 482-494 (2011).</li>
<li>Braune, A., Blaut, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Intestinal+bacterium+Eubacterium+cellulosolvens+deglycosylates+flavonoid+C-and+O-glucosides%5BTitle%5D)+AND+%22Appl+Environ+Microbiol%22%5BJournal%5D)">Intestinal bacterium Eubacterium cellulosolvens deglycosylates flavonoid C-and O-glucosides.</a> Appl Environ Microbiol. 78 (22), 8151-8153 (2012).</li>
<li>Hasanah, U., Miki, K., Nitoda, T., Kanzaki, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Aerobic+bioconversion+of+C-glycoside+mangiferin+into+its+aglycone+norathyriol+by+an+isolated+mouse+intestinal+bacterium%5BTitle%5D)+AND+%22Biosci+Biotechnol+Biochem%22%5BJournal%5D)">Aerobic bioconversion of C-glycoside mangiferin into its aglycone norathyriol by an isolated mouse intestinal bacterium.</a> Biosci Biotechnol Biochem. 85 (4), 989-997 (2021).</li>
<li>Kim, D. -H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Intestinal+bacterial+metabolism+of+flavonoids+and+its+relation+to+some+biological+activities%5BTitle%5D)+AND+%22Arch+Pharm+Res%22%5BJournal%5D)">Intestinal bacterial metabolism of flavonoids and its relation to some biological activities.</a> Arch Pharm Res. 21, 17-23 (1998).</li>
<li>Kim, M., Lee, J., Han, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Deglycosylation+of+isoflavone+C-glycosides+by+newly+isolated+human+intestinal+bacteria%5BTitle%5D)+AND+%22J+Sci+Food+Agric%22%5BJournal%5D)">Deglycosylation of isoflavone C-glycosides by newly isolated human intestinal bacteria.</a> J Sci Food Agric. 95 (9), 1925-1931 (2015).</li>
<li>Tao, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Isolation+and+identification+of+Enterococcus+gallinarum+P581a%2C+a+strain+of+intestinal+bacteria+deglycosylating+flavone+C-glycosides%5BTitle%5D)+AND+%22J+Gen+Appl+Microbiol%22%5BJournal%5D)">Isolation and identification of Enterococcus gallinarum P581a, a strain of intestinal bacteria deglycosylating flavone C-glycosides.</a> J Gen Appl Microbiol. 68 (3), 125-133 (2022).</li>
<li>Wang, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+newly+isolated+human+intestinal+strain+deglycosylating+flavonoid+C-glycosides%5BTitle%5D)+AND+%22Arch+Microbiol%22%5BJournal%5D)">A newly isolated human intestinal strain deglycosylating flavonoid C-glycosides.</a> Arch Microbiol. 204 (6), 310(2022).</li>
<li>Xu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Application+of+ultra-performance+liquid+chromatography+coupled+with+quadrupole+time-of-flight+mass+spectrometry+to+determine+the+metabolites+of+orientin+produced+by+human+intestinal+bacteria%5BTitle%5D)+AND+%22J+Chromatogr+B+Analyt+Technol+Biomed+Life+Sci%22%5BJournal%5D)">Application of ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry to determine the metabolites of orientin produced by human intestinal bacteria.</a> J Chromatogr B Analyt Technol Biomed Life Sci. 944, 123-127 (2014).</li>
<li>Zheng, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+newly+isolated+human+intestinal+bacterium+strain+capable+of+deglycosylating+flavone+C-glycosides+and+its+functional+properties%5BTitle%5D)+AND+%22Microb+Cell+Fact%22%5BJournal%5D)">A newly isolated human intestinal bacterium strain capable of deglycosylating flavone C-glycosides and its functional properties.</a> Microb Cell Fact. 18, 1-10 (2019).</li>
<li>Braune, A., Engst, W., Blaut, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Identification+and+functional+expression+of+genes+encoding+flavonoid+O-and+C-glycosidases+in+intestinal+bacteria%5BTitle%5D)+AND+%22Environ+Microbiol%22%5BJournal%5D)">Identification and functional expression of genes encoding flavonoid O-and C-glycosidases in intestinal bacteria.</a> Environ Microbiol. 18 (7), 2117-2129 (2016).</li>
<li>Kumano, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((FAD-dependent+C-glycoside-metabolizing+enzymes+in+microorganisms%3A+Screening%2C+characterization%2C+and+crystal+structure+analysis%5BTitle%5D)+AND+%22Proc.+Natl.+Acad.+Sci.+U.S.A%22%5BJournal%5D)">FAD-dependent C-glycoside-metabolizing enzymes in microorganisms: Screening, characterization, and crystal structure analysis.</a> Proc. Natl. Acad. Sci. U.S.A. 118 (40), e2106580118(2021).</li>
<li>Nakamura, K., Zhu, S., Komatsu, K., Hattori, M., Iwashima, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Deglycosylation+of+the+isoflavone+C-glucoside+puerarin+by+a+combination+of+two+recombinant+bacterial+enzymes+and+3-oxo-glucose%5BTitle%5D)+AND+%22Appl+Environ+Microbiol%22%5BJournal%5D)">Deglycosylation of the isoflavone C-glucoside puerarin by a combination of two recombinant bacterial enzymes and 3-oxo-glucose.</a> Appl Environ Microbiol. 86 (14), e00607-e00620 (2020).</li>
<li>He, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Structural+mechanism+of+a+dual-functional+enzyme+DgpA%2FB%2FC+as+both+a+C-glycoside+cleaving+enzyme+and+an+O-+to+C-glycoside+isomerase%5BTitle%5D)+AND+%22Acta+Pharm+Sin+B%22%5BJournal%5D)">Structural mechanism of a dual-functional enzyme DgpA/B/C as both a C-glycoside cleaving enzyme and an O- to C-glycoside isomerase.</a> Acta Pharm Sin B. 13 (1), 246-255 (2023).</li>
<li>Wang, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pro+isomerization+in+MLL1+PHD3-bromo+cassette+connects+H3K4me+readout+to+CyP33+and+HDAC-mediated+repression%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">Pro isomerization in MLL1 PHD3-bromo cassette connects H3K4me readout to CyP33 and HDAC-mediated repression.</a> Cell. 141 (7), 1183-1194 (2010).</li>
<li>Mossessova, E., Lima, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ulp1-SUMO+crystal+structure+and+genetic+analysis+reveal+conserved+interactions+and+a+regulatory+element+essential+for+cell+growth+in+yeast%5BTitle%5D)+AND+%22Mol+Cell%22%5BJournal%5D)">Ulp1-SUMO crystal structure and genetic analysis reveal conserved interactions and a regulatory element essential for cell growth in yeast.</a> Mol Cell. 5 (5), 865-876 (2000).</li>
<li>Delucas, L. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Protein+crystallization%3A+virtual+screening+and+optimization%5BTitle%5D)+AND+%22Prog+Biophys+Mol+Biol%22%5BJournal%5D)">Protein crystallization: virtual screening and optimization.</a> Prog Biophys Mol Biol. 88 (3), 285-309 (2005).</li>
<li>Minor, W., Cymborowski, M., Otwinowski, Z., Chruszcz, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((HKL-3000%3A+the+integration+of+data+reduction+and+structure+solution--from+diffraction+images+to+an+initial+model+in+minutes%5BTitle%5D)+AND+%22Acta+Crystallogr+D+Biol+Crystallogr%22%5BJournal%5D)">HKL-3000: the integration of data reduction and structure solution--from diffraction images to an initial model in minutes.</a> Acta Crystallogr D Biol Crystallogr. 62 (Pt 8), 859-866 (2006).</li>
<li>Li, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+efficient+C-glycoside+production+platform+enabled+by+rationally+tuning+the+chemoselectivity+of+glycosyltransferases%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">An efficient C-glycoside production platform enabled by rationally tuning the chemoselectivity of glycosyltransferases.</a> Nat Commun. 15 (1), 8893(2024).</li>
<li>Peters, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Characterization+of+membrane-bound+dehydrogenases+from+Gluconobacter+oxydans+621H+via+whole-cell+activity+assays+using+multideletion+strains%5BTitle%5D)+AND+%22Appl+Microbiol+Biotechnol%22%5BJournal%5D)">Characterization of membrane-bound dehydrogenases from Gluconobacter oxydans 621H via whole-cell activity assays using multideletion strains.</a> Appl Microbiol Biotechnol. 97 (14), 6397-6412 (2013).</li>
<li>Nasseri, S. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+alternative+broad-specificity+pathway+for+glycan+breakdown+in+bacteria%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">An alternative broad-specificity pathway for glycan breakdown in bacteria.</a> Nature. 631 (8019), 199-206 (2024).</li>
<li>Rose, J. P., Wang, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((SAD+phasing%3A+History%2C+current+impact+and+future+opportunities%5BTitle%5D)+AND+%22Arch+Biochem+Biophys%22%5BJournal%5D)">SAD phasing: History, current impact and future opportunities.</a> Arch Biochem Biophys. 602, 80-94 (2016).</li>
<li>Abramson, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Accurate+structure+prediction+of+biomolecular+interactions+with+AlphaFold+3%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Accurate structure prediction of biomolecular interactions with AlphaFold 3.</a> Nature. 630 (8016), 493-500 (2024).</li>
<li>Taberman, H., Parkkinen, T., Rouvinen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Structural+and+functional+features+of+the+NAD%28P%29+dependent+Gfo%2FIdh%2FMocA+protein+family+oxidoreductases%5BTitle%5D)+AND+%22Protein+Sci%22%5BJournal%5D)">Structural and functional features of the NAD(P) dependent Gfo/Idh/MocA protein family oxidoreductases.</a> Protein Sci. 25 (4), 778-786 (2016).</li>
<li>Jahn, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Understanding+the+chemistry+of+the+artificial+electron+acceptors+PES%2C+PMS%2C+DCPIP+and+Wurster%27s+Blue+in+methanol+dehydrogenase+assays%5BTitle%5D)+AND+%22J.+Biol.+Inorg.+Chem%22%5BJournal%5D)">Understanding the chemistry of the artificial electron acceptors PES, PMS, DCPIP and Wurster's Blue in methanol dehydrogenase assays.</a> J. Biol. Inorg. Chem. 25, 199-212 (2020).</li>
<li>Yu, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Biotransformation+of+puerarin+into+puerarin-6+%E2%80%B3-O-phosphate+by+Bacillus+cereus%5BTitle%5D)+AND+%22J+Ind+Microbiol+Biotechnol%22%5BJournal%5D)">Biotransformation of puerarin into puerarin-6 ″-O-phosphate by Bacillus cereus.</a> J Ind Microbiol Biotechnol. 39 (2), 299-305 (2012).</li>
<li>Lin, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Lysineless+HiBiT+and+NanoLuc+tagging+systems+as+alternative+tools+for+monitoring+targeted+protein+degradation%5BTitle%5D)+AND+%22ACS+Med+Chem+Lett%22%5BJournal%5D)">Lysineless HiBiT and NanoLuc tagging systems as alternative tools for monitoring targeted protein degradation.</a> ACS Med Chem Lett. 15 (8), 1367-1375 (2024).</li>
<li>Ma, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+Widespread+Radical-Mediated+Glycolysis+Pathway%5BTitle%5D)+AND+%22J+Am+Chem+Soc%22%5BJournal%5D)">A Widespread Radical-Mediated Glycolysis Pathway.</a> J Am Chem Soc. 146 (38), 26187-26197 (2024).</li>
<li>Yang, L., Lei, S., Xu, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rising+above%3A+exploring+the+therapeutic+potential+of+natural+product-based+compounds+in+human+cancer+treatment%5BTitle%5D)+AND+%22Tradit+Med+Res%22%5BJournal%5D)">Rising above: exploring the therapeutic potential of natural product-based compounds in human cancer treatment.</a> Tradit Med Res. 10 (3), 18(2025).</li>
<li>Zhu, C., Cook, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+concise+synthesis+of+%28%2B%29-artemisinin%5BTitle%5D)+AND+%22J+Am+Chem+Soc%22%5BJournal%5D)">A concise synthesis of (+)-artemisinin.</a> J Am Chem Soc. 134 (33), 13577-13579 (2012).</li>
<li>Schade, A. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((AKT+and+EZH2+inhibitors+kill+TNBCs+by+hijacking+mechanisms+of+involution%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">AKT and EZH2 inhibitors kill TNBCs by hijacking mechanisms of involution.</a> Nature. , 1-9 (2024).</li>
<li>Karpinska, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Selective+degradation+of+MLK3+by+a+novel+CEP1347-VHL-02+PROTAC+compound+limits+the+oncogenic+potential+of+TNBC%5BTitle%5D)+AND+%22J+Med+Chem%22%5BJournal%5D)">Selective degradation of MLK3 by a novel CEP1347-VHL-02 PROTAC compound limits the oncogenic potential of TNBC.</a> J Med Chem. 67 (17), 15012-15028 (2024).</li>
</ol>

<table><tbody><tr><td>2,6-dicloroindofenol (DCPIP)</td><td>Solarbio Ciência e Tecnologia</td><td>D6350</td><td></td></tr><tr><td>2-Mercaptoetanol(&beta;-ME)</td><td>Sigma-Aldrich</td><td>08168</td><td></td></tr><tr><td>&nbsp; 5 &vezes; Corante de Carga de Proteína</td><td>Sangon Biotech</td><td>C508320</td><td></td></tr><tr><td>Ácido Acético</td><td>Shanghai MACKLIN</td><td>A111247</td><td></td></tr><tr><td>Acetonitrila</td><td>Thermo Fisher Scientific&trade;? Invitrogen</td><td>A12147</td><td></td></tr><tr><td>Bruker Avance III HD 700 MHz Espectrômetro NMR</td><td>Bruker</td><td>AVANCE III HD 700MHz</td><td></td></tr><tr><td>Bruker Ultrashield 400 Plus 400MHz</td><td>Bruker</td><td>AVANCE III Centrífuga 400 MHz</td><td></td></tr><tr><td>5424R</td><td>Eppendorf</td><td>022620601</td><td></td></tr><tr><td>Chirascan&trade; plus CD espectrômetro</td><td>Applied Photophysics</td><td>Chirascan&trade; &nbsp;</td><td></td></tr><tr><td>Daidzein</td><td>Shanghai Yuanye Bio-Technology</td><td>B20227</td><td></td></tr><tr><td>Daidzin</td><td>Shanghai Yuanye Bio-Technology</td><td>B20226</td><td></td></tr><tr><td>Dithiothreitol (DTT)</td> <td>Solarbio Science and Technology</td><td>D8220</td><td></td></tr><tr><td>E. coli BL21 (DE3) células competentes</td><td>Thermo Fisher Scientific&trade;? Invitrogen</td><td>EC0114</td><td></td></tr><tr><td>Genisteína</td><td>Xangai Yuanye Bio-Tecnologia</td><td>S31565</td><td></td></tr><tr><td>Genisteína-8-C-glicosídeo</td><td>Xangai Yuanye Bio-Tecnologia</td><td>BP2095</td><td></td></tr><tr><td>Genistin</td><td>Xangai Yuanye Bio-Tecnologia</td><td>S31591</td><td></td></tr><tr><td>Glicose</td><td>Sigma-Aldrich</td><td>G8270</td><td></td></tr><tr><td>GraphPad Prisma 9</td><td>GraphPad Software</td><td>Versão? Prisma 9</td><td>Calcule os parâmetros cinéticos Km e kcat</td></tr><tr><td>HiLoad Superdex 200 pg colunas SEC preparativas</td><td>Cytiva</td><td>28989335</td><td></td></tr><tr><td>HisTrap HP Colunas de purificação de proteínas de etiqueta Cytiva</td><td></td><td>17524802</td><td></td></tr><tr><td>HiTrap Q FF coluna de cromatografia de troca aniônica</td><td>Cytiva</td><td>17515601</td><td></td></tr><tr><td>HKL-3000</td><td>HKL research Inc</td><td>&nbsp; versão 723</td><td></td></tr><tr><td>HPLC Coluna TC-C18(2), 170&Aring;, 5 &micro; m, 4,6 x 250 mm</td><td>Agilent Technologies</td><td>518925-902</td><td></td></tr><tr><td>Isopropil &beta;-D-1-tiogalactopiranosídeo? IPTG)</td><td>Sigma-Aldrich</td><td>PHG0010</td><td></td></tr><tr><td>Lactose</td><td>Sigma-Aldrich</td><td>17814</td><td></td></tr><tr><td>LC-20AT, cromatografia líquida de alta eficiência</td><td>Shimadzu</td><td>LC20AT</td><td></td></tr><tr><td>Maltose</td><td>Sigma-Aldrich</td><td>47288</td><td></td></tr><tr><td>Maltotriose</td><td>Sigma-Aldrich</td><td>443713</td><td></td></tr><tr><td>Cloreto de manganês</td><td>Shanghai Yuanye Bio-Technology</td><td>S31112</td><td></td></tr><tr><td>Metanol</td><td>Thermo Fisher Scientific&trade;? Invitrogen</td><td>M23142</td><td></td></tr><tr><td>NAD+</td><td>Solarbio Ciência e Tecnologia</td><td>N8110</td><td></td></tr><tr><td>Microscópio Estéreo Olympus SZX12</td><td>OLYMPUS&nbsp;</td><td>SZX7</td><td></td></tr><tr><td>fluoreto de fenilmetilsulfonil (PMSF)</td> <td>Sigma-Aldrich</td><td>P7626</td><td></td></tr><tr><td>Phenix</td><td>A equipe de desenvolvimento do Phenix</td><td>1.21.2</td><td>O Phenix é um pacote de software abrangente para determinação da estrutura cristalina de proteínas, incorporando métodos SAD (Dispersão Anômala de Comprimento de Onda Único) e MR (Substituição Molecular) com o software Phaser.</td></tr><tr><td>Cromatógrafo líquido preparativo de alto desempenho</td><td>Kit</td> de Triagem de Cristais de Proteína Waters <td>2545/2998</td><td></td></tr><tr><td></td><td>Hampton Research</td><td>HR2-110/HR2-112/HR2-144/HR2-126/HR2-098</td><td></td></tr><tr><td>Puerarin</td><td>Shanghai Yuanye Bio-Technology</td><td>B20446</td><td></td></tr><tr><td>Incubadora de agitação</td><td>Shanghai Zhichu Instrument Co.,Ltd</td><td>&nbsp; ZQZY-AF8</td><td></td></tr><tr><td>Pele de Cobra&trade; Membrana de diálise, 10 kDa MWCO, 35 mm</td><td>Thermo Fisher Scientific&trade;? Invitrogen</td><td>88245</td><td></td></tr><tr><td>Amido solúvel</td><td>Sigma-Aldrich</td><td>S9765</td><td></td></tr><tr><td>Sorvall LYNX 6000 Superspeed Centrifuge</td><td>Thermo Fisher Scientific&trade;</td><td></td><td></td>75006590</tr><tr><td>Leitor de Microplacas Multimodo SpectraMax i3x</td><td>DISPOSITIVOS MOLECULARES</td><td>SpectraMax i3x&nbsp;</td><td></td></tr><tr><td>Especificações do Espectrofotômetro de Microvolume SpectraMax QuickDrop</td><td>DISPOSITIVOS MOLECULARES</td><td>QuickDrop</td><td></td></tr><tr><td>Sacarose</td><td>Sigma-Aldrich</td><td>S5016</td><td></td></tr><tr><td>SunFire C18 OBD Prep Column</td><td>Waters</td><td>186002741</td><td></td></tr><tr><td>Thermo Fisher Scientific Q EXACTIVE</td><td>Thermo Fisher Scientific&trade;? Invitrogen</td><td>IQLAAEGAAPFALGMBDK</td><td></td></tr><tr><td>Protease 1 semelhante à ubiquitina</td><td>Thermo Fisher Scientific&trade;? Invitrogen</td><td>12588018</td><td></td></tr><tr><td>Freezer de temperatura ultra baixa</td><td>Thermo Fisher Scientific</td><td>905GP-ULTS</td><td></td></tr><tr><td>Concentrador centrífugo a vácuo</td><td>Beijing JM Technology</td><td>CV600</td><td></td></tr><tr><td>Vitexin</td><td>Sigma-Aldrich</td><td>49513</td><td></td></tr></tbody></table>

abordagens de biologia estrutural e química analítica para caracterização de enzimas metabólicas de c-glicosídeos na microbiota intestinal humana

Structural Biology and Analytical Chemistry Approaches for Characterizing C-Glycoside Metabolic Enzymes in Human Gut Microbiota

Seguindo os procedimentos descritos em Figura 1, primeiro expressamos os genes DgpA, DgpB e DgpC em E. coli BL21(DE3). Uma vez que DgpB e DgpC formam um complexo enzimático estável (complexo DgpB/C)10,26,27,34, as células que expressam essas duas proteínas foram combinadas para co-purificação. Após a lise celular, o sobrena...

Abordagens de Biologia Estrutural e Química Analítica para Caracterização de Enzimas Metabólicas de C-Glicosídeos na Microbiota Intestinal Humana

The gut microbiota has emerged as a key modulator of host health, particularly because of its capacity to metabolize dietary glycosides. While ...

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Biochemistry

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Um método específico e sensível para obter informações sobre o perfil de expressão de antígenos glicoesfingolipídeos em órgãos e células do sistema imunológico é descrito. O método tira partido da espectrometria de massa de iões armadilha permitindo passo a passo de fragmentação de moléculas de glicoesfingolipídeos para análise estrutural em comparação com padrões de síntese química.

Análise por espectrometria de massa de Antígenos glicoesfingolipídeos

Fonte: Hu, Y. et.al., Análise das Interações entre Endobióticos e Microbiota Intestinal Humana Usando Sistemas de Fermentação em Banhos In Vitro. J. Vis. Exp. (2019)Este vídeo demonstra a análise da produção de ácidos graxos de cadeia curta pela microbiota intestinal humana em resposta a exopolissacarídeos. Isso destaca a separação e quantificação dos produtos de fermentação microbiana usando cromatografia gasosa. Os resultados revelam que exopolissacarídeos modulam metabólitos microbianos que...

Investigando a Atividade Microbiana Intestinal Humana em Exopolissacarídeos

A comunidade bacteriana activa associada ao intestino de Spodoptera littoralis, foi determinada por-estável-sondagem isótopo (SIP) acoplado a pyrosequencing. Usando essa metodologia, a identificação das espécies de bactérias metabolicamente ativas dentro da comunidade foi feito com alta resolução e precisão.

Identificação de bactérias metabolicamente ativas no intestino do Generalista<em&gt; Spodoptera littoralis</em&gt; Via DNA Isótopos Estáveis ​​Sondagem Usando<sup&gt; 13</sup&gt; C-Glicose

Um ensaio de alto rendimento para triagem enzimática é descrito. Este kit de ensaio multiplexado pronto para uso é composto por substratos Chromogênicos Polymer Hydrogel (CPH) pré-escolhidos e substratos complexos de Chromogênico B iomassa (ICB). As enzimas-alvo são endoenzimas e proteases degradadoras de polissacarídeos.

High-throughput screening de enzimas Carboidratos-degradantes Usando Novel Insolúvel substrato cromogénico Kits de Ensaio

Aqui, apresentamos um protocolo para cultivo a microbiota do intestino do cólon in vitro, usando uma série de biorreatores que simulam as condições fisiológicas do tracto gastro-intestinal.

Aplicação avançada em Vitro cultivo tecnologia para estudar a Microbiota do intestino humano

Descrevemos aqui a elevado-throughput detecção e quantificação de oligossacarídeos de leite humano de fucosilado (HMOs) usando um biossensor de células inteiras. Também demonstramos aqui, a adaptação desta plataforma para análise de cepas de produção de HMO, com foco na melhoria da relação sinal / ruído.

Análise de fucosilado humanos leite trissacarídeos biotecnológicas contexto usando geneticamente codificado biosensores

Descrito aqui é um protocolo para investigar as interações entre endobióticos e microbiota intestinal humana usando sistemas in vitro de fermentação em lote.

Análise de interações entre endobióticos e microbiota intestinal humana usando sistemas de fermentação de banho in vitro

É descrito aqui um procedimento de funcionamento padrão simplificado para o perfilamento do microbioma usando o sequenciação metagenômica 16s rRNA e a análise usando ferramentas livremente disponíveis. Este protocolo ajudará os investigadores que são novos ao campo do microbioma assim como aqueles que exigem actualizações em métodos para conseguir o perfilamento bacteriano em uma definição mais elevada.

Análise de microbiota usando PCR de duas etapas e sequenciamento do gene 16S rRNA de próxima geração

A busca aberta permite identificar glycopeptídeos decorados com composições glican até então desconhecidas. Dentro deste artigo, uma abordagem simplificada para realizar buscas abertas e subsequentes pesquisas de gliocopeptídio focadas em glycan são apresentadas para amostras bacterianas usando Acinetobacter baumannii como modelo.

A aplicação de abordagens baseadas em pesquisa aberta para a identificação de Glycopeptides ligados ao Acinetobacter baumannii O

Este estudo estabelece um conjunto de procedimentos operacionais padrão (SOPs) para rastrear e isolar com eficiência as bactérias intestinais capazes de clivar os glicosídeos C.

Triagem e isolamento de bactérias intestinais clivadas com glicosídeo C

Aprenda a caracterizar enzimas metabólicas de C-glicosídeos da microbiota intestinal humana usando cristalografia de proteínas, LC-MS/MS e RNM. Este método integra biologia estrutural e química analítica.

Abordagens de Biologia Estrutural e Química Analítica para Caracterização de Enzimas Metabólicas de C-Glicosídeos na Microbiota Intestinal Humana (Vídeo) | JoVE Journal

Abordagens de Biologia Estrutural e Química Analítica para Caracterização de Enzimas Metabólicas de C-Glicosídeos na Microbiota Intestinal Humana

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