产耐热α-葡萄糖苷酶重组大肠杆菌的构建及发酵条件优化

Construction of Recombinant E.coli for Thermostable α-Glucosidase Production and Fermentation Conditions Optimization

以非解朊栖热菌HG102基因文库筛选的重组质粒pUC18-atgly为模板PCR扩增α-葡萄糖苷酶结构基因,与表达载体pET28α(+)连接,转入大肠杆菌BL21(DE3),经异丙基硫代-β-D-半乳糖苷(IPTG)诱导表达.对重组菌的产酶条件进行了初步优化,最终酶活由4.25U/mL提高到40.85U/mL.

α-Glucosidase gene was multiplied by PCR with recombinant plasmid pUC18-atgly from genomic library of thermophilic eubacterium Thermus nonproteolyticus HG102 as template,which was ligated with plasmid pET28α（＋） and transformed into E.coli BL21.IPTG induced the expression of α-glucosidase.The α-glucosidase producing conditions were optimized preliminarily.The final enzyme activity increased from 4.25 U/mL to 40.85 U/mL.