两株蜡样芽胞杆菌α-淀粉酶差异性分析及基因的克隆和表达  

Difference Analysis of Two a-amylase from Bacillus cereus and Their Gene Cloning and Expression

作　　者：张琳琳[1] 王琦[1] 李荣禧[2] 王勇军[1] 梅汝鸿[1] 

机构地区：[1]中国农业大学植物病理学系,北京100094 [2]内蒙古农业大学农学院,呼和浩特010018

出　　处：《农业生物技术学报》2008年第4期676-680,共5页Journal of Agricultural Biotechnology

基　　金：国家高新技术研究与发展计划(863)项目(No.2006AA10A211);苹果轮纹病菌及其抗性资源信息库的建立(农业公益性行业科技)项目(No.nyhyzx07-055)资助。

摘　　要：蜡样芽胞杆菌(Bacillus cereus)是产生具有重要工业生产价值的耐高温α-淀粉酶的优良菌株。对2株蜡样芽胞杆菌(B905和B904)分泌的α-淀粉酶酶学性质的研究显示,淀粉酶的活性稳定温度在90～100℃,耐热性和酶活不依赖Ca^(2+),但淀粉酶的分子量大小有明显的不同。PCR方法克隆得到了两种耐高温淀粉酶的基因全长(amy905和amy904),并在大肠杆菌诱导表达。结果显示,amy904基因长度为1362bp,其蛋白分子量约为55 kD;amy905基因长度为1 761 bp,其蛋白分子量约为68 kD。Bacillus cereus can produce α-amylase which has important industrial production value and can endure high temperature. The enzymatic characteristic of two α-amylase from B. cereus （B905 and B904） were studied. And the results showed that the activity stability temperature of both α-amylase was 90-100℃, and their thermal stabilities and enzymatic activities did not depend on Ca^2＋. But their protein molecular weights were obviously different. The structural genes of amy905 and amy904 were successfully cloned by PCR and expressed in Escherichia coli. And the results showed that the amy904 and amy905 genes were 1 362 bp and 1 761 bp long and their protein molecular weights were about 55 kD and 68 kD, respectively.

关 键 词：蜡样芽胞杆菌 淀粉酶 基因克隆 表达 

分 类 号：S188[农业科学—农业基础科学]