铜绿假单胞菌脂肪酶基因在枯草芽孢杆菌中的克隆与表达  

Cloning and Expression of Pseudomonas aeruginosa Lipase Gene in Bacillus subtilis

作　　者：韩振林[1] 朱传合[1] 赵明明[1] 赵玉金[1] 路福平[1] 戚薇[1] 杜连祥[1] 

机构地区：[1]天津市工业微生物重点实验室天津科技大学生物工程学院

出　　处：《食品与发酵工业》2006年第1期11-14,共4页Food and Fermentation Industries

基　　金：天津科技大学校重点基金(20040101);天津市自然基金(05YFJMJCO2800)资助

摘　　要：利用PCR方法从分泌脂肪酶的铜绿假单胞菌基因组DNA中扩增得到了脂肪酶基因,并测定其核苷酸序列,利用基因重组技术构建了脂肪酶基因的分泌表达载体,并在枯草芽孢杆菌中进行了分泌表达。SDS聚丙烯酰胺凝胶电泳表明,表达蛋白占发酵液中蛋白的25%,采用NaOH碱滴定法测定其发酵液酶活为15.26U/mL。DNA fragment encoding lipase was amplified by PCR from genomic DNA , and cloned into pWb980 vector. The secretion/expression vector with lipase gene was constructed by gene recombination technology and high level expression of lipase in Bacillus subtilis DB104 was obtained. It was seen that the expression protein was about 25 % of the total proteins by SDS-PAGE and the lipase in supernatant had the activity of hydrolyzing olive oil, which was 15.26u/mL assayed by NaOH alkaline titration method.

关 键 词：脂肪酶 枯草芽孢杆菌 克隆 表达 

分 类 号：Q78[生物学—分子生物学]