Production of an oligosaccharide-specific cellobiohydrolase from the thermophilic fungus Thielavia terrestris |
| |
| Authors: | James Sy-Keen Woon Mukram Mohamed Mackeen Amirul Husna bin Sudin Nor Muhammad Mahadi Rosli Md Illias Abdul Munir Abdul Murad Farah Diba Abu Bakar |
| |
| Affiliation: | 1.School of Biosciences and Biotechnology, Faculty of Science and Technology,Universiti Kebangsaan Malaysia,Bangi,Malaysia;2.School of Chemical Sciences and Food Technology, Faculty of Science and Technology,Universiti Kebangsaan Malaysia,Bangi,Malaysia;3.Institute of Systems Biology (INBIOSIS),Universiti Kebangsaan Malaysia,Bangi,Malaysia;4.Malaysia Genome Institute,Kajang,Malaysia;5.Department of Bioprocess Engineering, Faculty of Chemical Engineering,Universiti Teknologi Malaysia,Skudai,Malaysia |
| |
| Abstract: | ObjectivesTo express and determine the hydrolytic activity of a cellobiohydrolase (TTCBH6B) from the thermophilic fungus Thielavia terrestris in Pichia pastoris.Results Ttcbh6B encodes a protein of 507 amino acid residues with a predicted molecular mass of 54 kDa. TTCBH6B contains a familial 6-glycosyl hydrolase catalytic domain and a type I carbohydrate-binding module. TTCBH6B was expressed and purified to homogeneity but the purified enzyme was inactive against Avicel. It could, however, digest Celluclast-treated Avicel producing cellobiose (0.27 μmol min?1 mg?1). To determine the substrate preferences of TTCBH6B, oligosaccharides of varying numbers of subunits were generated by acid hydrolysis of Avicel and fluorescently tagged. Peaks corresponding to oligosaccharides containing three to six glucose units were reduced to cellobiose after addition of TTCBH6B.ConclusionTTCBH6B is active against shorter oligosaccharides rather than polysaccharides. |
| |
| Keywords: | |
| 本文献已被 SpringerLink 等数据库收录！ |
|
