Biochemical property of an endo-glucanase-like enzyme from Clostridium sp. Z-7026
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Title Biochemical property of an endo-glucanase-like enzyme from Clostridium sp. Z-7026
Creator Paripok Phitsuwan
Contributor Sengthong Lee, Salita Eiamboonsert, Khanok Ratanakhanokchai
Publisher The Thai Society for Biotechnology
Publication Year 2562
Keyword Cellulose, Cello-Oligosaccharide, Clostridium species, Endoglucanase
Abstract 1. Clostridium sp. Z-7026 is a cellulose-degrading anaerobic bacterium, and its genome was recently sequenced. By Blast analysis against the NCBI protein database, one gene encoding a cellulase-like protein was identified. This deduced protein was named Cel_2759, and the Cel_2759 encoding gene was cloned, expressed in Escherichia coli, and purified. The purified recombinant protein had an estimated size of 107 kDa. To reveal its hydrolyzing ability, five substrates, namely carboxymethylcellulose (CMC), regenerate amorphous cellulose (RAC), crystalline cellulose Avicel beechwood xylan (BWX), and pretreated rice straw were used for activity assay. Results showed that, with in 15 min, Cel_2759 actively hydrolyzed CMC, yielding a reducing sugar concentration of 689.45 ?g/mL. However, activities against RAC, BWX and rice straw required pronged incubation time up to 16 h, yielding reducing sugar concentrations of 73.32 ?g/mL, 1393.05 ?g/mL, 875.704 ?g/mL, respectively. At 24 h, activity against Avicel was not observed, suggesting that Cel_2759 was unable to hydrolyze crystalline cellulose
2. by contrast, it preferred soluble long chain substrate CMC for hydrolysis. Using CMC as the substrate, Cel_2759 had its own optimal pH and temperature for its biological function at pH 7.0-8.0 and 55 ?C. The hydrolysis products from CMC by Cel_2759 were a mixture of cellooligomers, suggesting that Cel_2759 was an endo-acting enzyme.
Language EN
URL Website http://tsb2019.com/
Website title The 31st Annual Meeting of the Thai Society for Biotechnology and International Conference (TSB 2019)
Thai Society for Biotechnology

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