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Description

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.

Publisher Location

Las Vegas (Nev.)

Publication Date

12-9-2022

Publisher

University of Nevada, Las Vegas

Language

English

Controlled Subject

Waste products as fuel

Disciplines

Cell and Developmental Biology | Environmental Health | Integrative Biology

File Format

pdf

File Size

376 KB

Comments

Faculty Mentors:Brian P. Hedlund, Marike Palmer

Rights

IN COPYRIGHT. For more information about this rights statement, please visit http://rightsstatements.org/vocab/InC/1.0/

Characterization of the thermophilic xlanase Fsa272 from Candidatus Fervidibacter sacchari belonging to glycoside hydrolase family GH10


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