title: Expression of a secretory β-glucosidase from Trichoderma reesei in Pichia pastoris and its characterization.
abstract: A β-glucosidase gene (bglI) from Trichoderma reesei was cloned into the pPIC9 vector and integrated into the genome of Pichia pastoris GS115. Under the control of the methanol-inducible alcohol oxidase (AOX) promoter and using Saccharomyces cerevisiae secretory signal peptide (α-factor), the recombinant β-glucosidase was expressed and secreted into the culture medium. The maximum recombinant β-glucosidase activity achieved was 60 U/ml, and β-glucosidase expression reached 0.3 mg/ml. The recombinant 76 kDa β-glucosidase was purified 1.8-fold with 26% yield and a specific activity of 197 U/mg. It was optimally active at 70 °C and pH 5.0.
authors.affiliation: National Engineering Laboratory for High-efficiency Enzyme Expression, Fuzhou, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Chen
authors.forename: Xianyu
authors.initials: X
authors.lastname: Fu
authors.forename: Tzi Bun
authors.initials: TB
authors.lastname: Ng
authors.forename: Xiu-Yun
authors.initials: XY
authors.lastname: Ye
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D004789:Enzyme Activation; D004795:Enzyme Stability; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics; D001617:beta-Glucosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D001617:beta-Glucosidase