酸性α-淀粉酶基因在枯草芽孢杆菌中的高效表达  

High-level Expression of Acid-stable α-amylase Gene of Bacillus amyloliquefaciens in Bacillus subtilis WB600

作　　者：王彩澜[1] 刘新育[1] 李学琴[1] 刘亮伟[1] 陈红歌[1] 

机构地区：[1]河南农业大学生命科学学院农业部农业微生物酶工程重点实验室,河南郑州450002

出　　处：《食品与发酵工业》2012年第5期52-55,共4页Food and Fermentation Industries

基　　金：河南省教育厅自然科学研究计划(2010A180006)

摘　　要：酸性α-淀粉酶是发酵行业用量最大的酶类,为了实现酸性α-淀粉酶基因的高效表达,将本实验室已经克隆得到的去信号肽的酸性α-淀粉酶基因在枯草芽孢杆菌WB600宿主中进行表达,成功的构建了表达菌株pHT43-amy/WB600。在初始菌浓度OD600为0.8时加入终浓度为0.9 mmol/L的IPTG,诱导6 h的条件下测得酶活力为1 230 U/mL,又由于宿主菌WB600外分泌蛋白较少,因此具有明显的生产优势。Acid-stable α-amylase is widely used in fermentation industry. In order to highly express acid-stable or-amylase gene, a recombinant vector containing no signal peptide of acid-stable or-amylase gene from Bacillus amyloliquefaciens was constructed followed by transformation into Bacillus subtilis WB6oo host. The highest activity of recombinant acid-stable α-amylase was 1230 U/mL under the condition as follows： OD600 of the initial bacteria was 0.8 and final concentration of IPTG was 0.9 mmol/L, and the cell suspension was cultured for 6 hours after addition of IPTG. As the recombinant Bacillus subtilis WB600 secreted ylase, it showed obvious advantages in amylase production. few other proteins except the targeted acid-stableα-am-ylase, it showed obvious advantages in amylase production.

关 键 词：酸性α-淀粉酶基因 高效表达 枯草芽孢杆菌 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]