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OALib Journal期刊
ISSN: 2333-9721
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植物乳杆菌bbe7的胆盐水解酶基因在毕赤酵母中的分泌表达
DOI: 10.3724/SP.J.1145.2014.00107, PP. 107-111
Keywords: 植物乳杆菌bbe7,胆盐水解酶,毕赤酵母,分泌表达,正交试验设计
Abstract:
由于用大肠杆菌对植物乳杆菌来源的bsh基因进行胞内和胞外表达时，它们很容易形成包涵体，而且大肠杆菌的内毒素会显限制bsh的应用，所以利用毕赤酵母这一高效的表达系统对其进行了分泌表达.首先用融合pcr的方法将信号肽α-factor与bsh连接，然后再克隆到质粒ppic9k上，得到重组质粒ppic9k-α-factor-bsh.重组质粒经过salⅰ线性化后转化至毕赤酵母gs115.表达产物经sds-page分析和酶活测定发现，重组bsh的相对分子质量（mr）和胞外酶活分别为37.0×103和1.08uml-1.通过正交实验[l9（34）]对发酵条件优化后，胞外总酶活达到2.69uml-1，比原来提高了1.49倍.研究为胆盐水解酶的分离纯化、酶学性质研究以及大规模生产奠定了一定的基础.
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