具有辅助降解纤维素功能的大斑刚毛座腔菌糖苷水解酶GH61的鉴定、异源表达及功能分析

马玉倩, 孙东辉, 岳浩峰, 辛佳瑜, 刘宁, 曹志艳

Identification, Heterologous Expression and Functional Analysis of a GH61 Family Glycoside Hydrolase from Setosphaeria turcica with the Assisting Function in Degrading Cellulose

MA Yu-qian, SUN Dong-hui, YUE Hao-feng, XIN Jia-yu, LIU Ning, CAO Zhi-yan

图3 StGH61-11重组蛋白异源表达及优化诱导条件 A：目的片段StGH61-11的PCR扩增产物琼脂糖凝胶电泳结果, 1-3：StGH61-11的cDNA片段, M：DNA marker；B：PCR验证重组转化子的琼脂糖凝胶电泳结果, 1-3：Pet32a-StGH61-11 重组质粒, M：DNA marker；C：SDS-PAGE 检测pET32a-StGH61-11（BL21）重组菌表达蛋白, 0：Pet32a-StGH61-11-BL21（DE3）重组菌未诱导对照, 1 ：Pet32a-StGH61-11-BL21（DE3）重组菌经IPTG诱导后, M ：预染蛋白分子量标准；D：诱导时间对重组蛋白StGH61-11酶活的影响；E：IPTG浓度对重组蛋白StGH61-11酶活的影响

Fig. 3 Heterologous expression of StGH61-11 recombinant protein and optimized induction conditions A: Agarose gel electrophoresis results of the target gene StGH61-11 PCR products, 1-3: GH61-11 cDNA fragment. M：DNA marker. B: Agarose gel electrophoresis results of PCR for recombinant transformants, 1-3: Pet32a-StGH61-11 recombinant plasmid, M：DNA marker. C: SDS-PAGE detection of whole bacterial lysate expressed by recombinant pET32a-StGH61-11（BL21）, 0 : Pet32a-StGH61-11-BL21（DE3）not induce control, 1 : Pet32a-StGH61-11-BL21（DE3）the recombinant bacteria were induced by IPTG, M : Standard for molecular weight of pre-dyed protein. D: Effect of induction time on the enzymatic activity of recombinant protein StGH61-11. E: Effect of IPTG concentration on the enzymatic activity of recombinant protein StGH61-11