National Engineering Laboratory for High-efficiency Enzyme Expression, Fuzhou, China.
Biotechnol Lett. 2011 Dec;33(12):2475-9. doi: 10.1007/s10529-011-0724-3. Epub 2011 Aug 9.
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.
里氏木霉的β-葡萄糖苷酶基因（bglI）被克隆到 pPIC9 载体中，并整合到毕赤酵母 GS115 的基因组中。在甲醇诱导的醇氧化酶（AOX）启动子的控制下，利用酿酒酵母分泌信号肽（α-因子），重组β-葡萄糖苷酶被表达并分泌到培养基中。最大的重组β-葡萄糖苷酶活性达到 60 U/ml，β-葡萄糖苷酶表达量达到 0.3 mg/ml。重组的 76 kDaβ-葡萄糖苷酶经纯化后，酶活回收率为 26%，比活为 197 U/mg，提高了 1.8 倍。其最适活性温度为 70°C，最适 pH 值为 5.0。
