大肠杆菌植酸酶基因appA的克隆与高效表达  

Cloning and Overexpression of Phytase Gene appA form Escherichia coli

作　　者：陈寅[1] 朱中泽[1] 张志芳[1] 何家禄[1] 

机构地区：[1]中国农业科学院蚕业研究所农业部家蚕生物技术重点开放实验室,镇江212018

出　　处：《微生物学通报》2004年第3期74-78,共5页Microbiology China

基　　金：江苏省高技术研究计划资助项目(No.BG2002313)

摘　　要：从猪粪便中分离并筛选出高效生产酸性植酸酶和磷酸酶双功酶(appA植酸酶)的大肠杆菌菌株。通过PCR方法从该菌株基因组中扩增获得了植酸酶基因appA,测序结果显示该基因编码区全长1,299个核苷酸。将该基因克隆到原核表达载体pET-28a(+)上,通过转化的大肠杆菌BL21在试管摇床培养条件下得到了高效表达,其表达量达到692U/mL。酶学特性分析表明其反应的最适pH为4.5,最适温度为60℃。A strain Escherichia coli with high production of phytase was screened from pig excreta. Phytase gene appA, with 1,299 bp coding region in full length, was cloned from its genome by polymerase chain reaction (PCR) . The gene appA was then cloned into the prokaryotic expression vector pET-28a ( + ) . In the host BL21, the phytase appA was overexpressed by shaker-cultivation (up to 692 U/mL) . The enzymatic analysis of the prokaryotic derived appA phytase revealed that its optimal pH and temperature was 4.5 and 60℃, respectively.

关 键 词：大肠杆菌 植酸酶 APPA 基因表达 

分 类 号：S816.1[农业科学—饲料科学]