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Expression of a secretory β-glucosidase from Trichoderma reesei in Pichia pastoris and its characterization

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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.

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Authors and Affiliations

  1. National Engineering Laboratory for High-efficiency Enzyme Expression, Fuzhou, China

    Ping Chen, Xianyu Fu & Xiu-Yun Ye

  2. College of Biological Science and Technology, Fuzhou University, Fuzhou, China

    Xiu-Yun Ye

  3. School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China

    Tzi Bun Ng

  4. 3/f, Building No. 3, No. 7, JinZhou Road North, JinShan Industrial Zone, Fuzhou, Fujian, 350002, People’s Republic of China

    Xiu-Yun Ye

Authors
  1. Ping Chen
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  2. Xianyu Fu
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  3. Tzi Bun Ng
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  4. Xiu-Yun Ye
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Correspondence to Xiu-Yun Ye.

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Chen, P., Fu, X., Ng, T.B. et al. Expression of a secretory β-glucosidase from Trichoderma reesei in Pichia pastoris and its characterization. Biotechnol Lett 33, 2475–2479 (2011). https://doi.org/10.1007/s10529-011-0724-3

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  • DOI: https://doi.org/10.1007/s10529-011-0724-3

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