多粘芽孢杆菌 (Bacillus polymyxa) β-葡萄糖苷酶基因在大肠杆菌中的表达、纯化及酶学性质分析  

Expression,Purification and Enzymatic Characterization of Bacillus polymyxa β-glucosidase Gene(bglA) in Escherichia coli

作　　者：赵云[1] 刘伟丰[1] 毛爱军[1] 江宁[1] 董志扬[1] 

机构地区：[1]中国科学院微生物研究所,北京100080

出　　处：《生物工程学报》2004年第5期741-744,共4页Chinese Journal of Biotechnology

基　　金：国家高技术研究发展计划项目 (No .2 0 0 1AA2 14 15 1);中科院知识创新方向性课题 (No .KJCX2-SW-2 0 6-1)~~

摘　　要：从多粘芽孢杆菌 (Bacilluspolymyxa 1 794 )中克隆得到 β_葡萄糖苷酶基因bglA。将其构建在大肠杆菌 (Es cherichiacoli)表达载体pET2 8a(+)上 ,转化E .coliBL2 1,获得重组工程菌BL1979。重组表达的 β_葡萄糖苷酶的酶活力达到 2 4 7IU mL ,经镍柱纯化后的 β_葡萄糖苷酶最适温度为 37℃ ,最适pH值为 7 0 ,该酶经纯化后纯度可达92 7%。用非变性梯度聚丙烯凝胶电泳发现该酶具有多种寡聚体形式 ,经荧光底物活性染色表明这些寡聚体均具有The β-glucosidase encoding gene bglA was cloned from Bacillus polymyxa 1 794 The bglA gene was inserted in expression vector pET28a(+) and transformed into Escherichia coli BL21 (DE3), finally the recombinant strain BL1979 was obtained. Induced by IPTG, the expression β-glucosidase activity reached to 24 7 IU/mL. The optimum temperature and optimum pH of the recombinant expression β-glucosidase in BL1979 were 37℃ and 7 0 respectively,the purity can reach to 92 7%. Analysis of the fusion protein by nondenaturing gradient gel electrophoresis,we found the fusion protein exists in dimmer,tetramer,hexamer and octamer, they all have hydrolase activity.

关 键 词：Β-葡萄糖苷酶 多粘芽孢杆菌 重组 表达 克隆 

分 类 号：Q786[生物学—分子生物学]