Characterization of the thermophilic xlanase Fsa272 from Candidatus Fervidibacter sacchari belonging to glycoside hydrolase family GH10
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Candidatus Fervidibacter sacchari is a novel, facultatively anaerobic, hyperthermophilic bacterium found in terrestrial geothermal springs globally. Its genome encodes 115 putative glycoside hydrolase enzymes that are predicted to hydrolyze glycosidic bonds between carbohydrates. Fsa272, a member of the glycoside hydrolase family 10, was synthesized and cloned into Escherichia coli strain T7 Express. The transformed E. coli was grown with LB broth and ampicillin at 37°C. Fsa272 expression was induced with isopropylthio-beta-galactoside (IPTG), and the lysate was heat purified for 15 minutes at 80° C. The 3,5-dinitrosalicylic acid assay identified xylanase activity with a pH range of 4.5 to 10.5 (pHopt 5.5) and a temperature range of 60 to 90°C (Topt 80-90°C). The para-nitrophenol assay was used to determine the Michaelis-Menten kinetic parameters of Fsa272, resulting in KM of 1.8 mM and V max of 232.6 μM/min. The characterization of Fsa272 provides critical information on Ca. F. sacchari and its potential application in converting polysaccharide waste to biofuels.
Las Vegas (Nev.)
University of Nevada, Las Vegas
Waste products as fuel
Cell and Developmental Biology | Environmental Health | Integrative Biology
Torosian, Nicole; Covington, Jonathan K.; Cook, Allison M.; Nou, Nancy O.; and Cheng, Jan-Fang, "Characterization of the thermophilic xlanase Fsa272 from Candidatus Fervidibacter sacchari belonging to glycoside hydrolase family GH10" (2022). Undergraduate Research Symposium Posters. 126.
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