Expression of a secretory β-glucosidase from Trichoderma reesei in Pichia pastoris and its characterization
| Autor: | Xiuyun Ye, Tzi Bun Ng, Fu Xianyu, Ping Chen |
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| Rok vydání: | 2011 |
| Předmět: |
Signal peptide
Saccharomyces cerevisiae Bioengineering Applied Microbiology and Biotechnology Pichia law.invention Pichia pastoris law Enzyme Stability Cloning Molecular Trichoderma reesei Trichoderma biology business.industry Chemistry beta-Glucosidase General Medicine biology.organism_classification Recombinant Proteins Alcohol oxidase Biotechnology Enzyme Activation Biochemistry Recombinant DNA business |
| Zdroj: | Biotechnology Letters. 33:2475-2479 |
| ISSN: | 1573-6776 0141-5492 |
| DOI: | 10.1007/s10529-011-0724-3 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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