Bacillusa myloliquefaciens中性植酸酶基因的原核表达及蛋白纯化和性质  

Expression of Neutral Phytase Gene from Bacillus amyloliquefaciens in Escherichia coli

作　　者：陈艳[1] 孙建义[1] 赵学新 付石军 翁晓燕[3] 

机构地区：[1]浙江大学教育部动物分子营养学重点实验室,浙江杭州310029 [2]山东省潍坊市畜牧技术推广站,山东潍坊261000 [3]浙江大学生命科学学院,浙江杭州310029

出　　处：《食品与生物技术学报》2005年第2期60-64,72,共6页Journal of Food Science and Biotechnology

摘　　要：淀粉液化芽孢杆菌Bacillus amyloliquefaciens 是一种嗜热细菌,其产生的中性植酸酶具有很好的应用前景.通过TD PCR技术将Bacillus amyloliquefaciens 编码的中性植酸酶基因克隆至原核表达载体pET 30a(+)上,在大肠杆菌 BL21(DE3)中得到了高效表达,表达量占大肠杆菌可溶性蛋白的33.5%.采用金属Ni2+亲和层析对基因表达产物进行纯化,表达产物具有正常的生物学功能.Bacillus amyloliquefaciens is one of the few thermophilic bacteria. The phytase found in B. amyloliquefaciens has shown the extremely thermostablity and considerable potential in large-scale production. The gene of phytase BPhyC from B. amyloliquefaciens was amplified by TD-PCR, inserted into the plasmid pET-30a(+), and fused with the 6 His tag. The recombinant plasmid was transformed to E.coli BL21 (DE3). The recombinant protein was purified by heating and immobilizing metal affinity chromatography. The expression phytase was over 40% the total soluble protein of E.coli and has normal bioactivity.

关 键 词：Bacillus 原核表达 植酸酶基因 蛋白纯化 中性植酸酶 性质 基因表达产物 大肠杆菌 PCR技术 NI^2+ 可溶性蛋白 生物学功能 芽孢杆菌 淀粉液化 嗜热细菌 基因克隆 高效表达 亲和层析 表达量 

分 类 号：Q786[生物学—分子生物学]