title: High activity cellulase production by recombinant Trichoderma reesei ZU-02 with the enhanced cellobiohydrolase production.
abstract: The cbh1 strong promoter was employed to over-express the cbh2 gene for enhancing cellobiohydrolase (CBH) production in Trichoderma reesei because CBH II component has higher specific activity than CBH I and is an important component in cellulase. The recombinant plasmid pCAMBIA1300-hph-PsCT containing strong expression cassette was constructed and transformed into T. reesei via optimized Agrobacterium-mediated transformation, producing 324 positive T. reesei transformants for the two steps of screening. Ten fast-growing T. reesei transformants were selected, amongst which C10 was found to have the highest filter paper activity 28.92±2.45 IU/mL, 4.3-fold higher than that of ZU-02, 6.71±0.79 IU/mL. C10 also has the highest cellobiohydrolase activity 122.44±7.42 U/mL, 5.4 times higher than that of ZU-02, 22.49±2.27 U/mL. The cellulase from C10 performed better (93.06±2.83%) than the one from ZU-02 in enzymatic hydrolysis because the exo-exo-synergism played a role.
authors.affiliation: Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Fang
authors.forename: Liming
authors.initials: L
authors.lastname: Xia
mesh_terms: D060054:Agrobacterium / Q000378:metabolism; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000096:biosynthesis*; D006868:Hydrolysis; D010957:Plasmids / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics*; D014170:Transformation, Genetic; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics*
chemical_list: D043366:Cellulose 1,4-beta-Cellobiosidase
keywords: Cellobiohydrolase; Cellulase; Enhanced expression; Enzymatic hydrolysis of corn stover; Recombinant Trichoderma reesei