多角体蛋白对植酸酶的包埋  

Immobilization of Phytase by Polyhedra of BmCPV

作　　者：郭建军[1] 袁林[1] 曾静[1] 付锦楠[1] 魏国汶[1] 杨一兵[1] 

机构地区：[1]江西省科学院微生物研究所,南昌330096

出　　处：《江西科学》2016年第2期182-186,共5页Jiangxi Science

基　　金：江西省重大科技创新研究项目"多角体蛋白对植酸酶的包裹技术及其应用研究"(20124ACF01100);江西省科学院对外合作项目"昆虫细胞内进行猪干扰素的多角体包裹研究"(H2014004)

摘　　要：植酸酶是水解植酸及其盐类生成肌醇和磷酸的一类酶的总称。作为一种新型饲料添加剂,植酸酶在动物营养及环境保护等领域具有很大的应用潜力。为了获得高活性的植酸酶app A,首先将app A基因克隆到载体p Fast Bac Dual中,构建转移载体p Fast Bac Dual-polyh-app A;然后转化大肠杆菌Escherichia coli DH10Bac感受态细胞;通过位点特异性转座,将app A基因整合到Bacmid穿梭载体中,构建表达质粒Bacmid:Ac-polyh/app A。最后,通过脂质体将表达质粒转染Sf9昆虫细胞。Western blotting和酶活测定法对app A的表达和活性进行分析,结果表明在昆虫细胞中app A酶表达成功,植酸酶能够被包埋进多角体内部,并具有较好的植酸酶活性,为植酸酶大规模生产和应用提供了可参考利用的技术。Phytase is a kind of enzymes that stepwise hydrolyse phytic acid to release phosphate radicals and inositol derivatives. As a new type of feed additives,it has great application potential in the areas of animal nutrition and environmental protection. In this study,app A enzyme was expressed in the baculovirus system and its activity was tested. We inserted app A gene into the p Fast Bac Dual,a baculovirus transfer vector and then transformed it into DH10 Bac containing a shuttle vector of Bacmid. After co-transfecting the recombinant plasmid into Sf9 insect cells,the expression of phytase was visualized by SDS-PAGE and Western blotting,the results showed the app A protein could be incorporated into the polyhedra and has good enzymatic activitiy. High-level expression of phytase with high specific activity is an effective way to improve phytase fermentation potency and reduce its production cost.

关 键 词：植酸酶 多角体 包埋 

分 类 号：S884.5[农业科学—特种经济动物饲养]