不同表达模式组合对谷氨酸棒状杆菌中脑钠肽蛋白表达的影响

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  • 1(江南大学 生物工程学院,江苏 无锡,214122) 2(江南大学,粮食发酵工艺与技术国家工程实验室,江苏 无锡,214122)3(江南大学,工业生物技术教育部重点实验室,江苏 无锡,214122)

网络出版日期: 2018-09-13

基金资助

国家自然科学基金(31570034);江苏省自然科学基金(BK20150148);江苏省大学生创新训练计划(2017102950 46X)

The effect of different expression patterns on B-type natriuretic peptide expression in Corynebacterium glutamicum

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  • 1(School of Biotechnology, Jiangnan University, Wuxi 214122, China)2(National Engineering Laboratory of Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China)3(Key Laboratory of Industrial Biotechnology, Jiangnan University, Wuxi 214122, China)

Online published: 2018-09-13

摘要

外源蛋白表达系统按照启动子作用方式可分为诱导型和组成型,按照蛋白表达后的定位可分为胞内表达和分泌表达。为了研究外源蛋白在谷氨酸棒状杆菌中最适的表达模式组合,以绿色荧光蛋白(green fluorescent protein, GFP)为模式蛋白构建用于研究谷氨酸棒状杆菌不同表达模式的快速表达体系,并利用这一体系研究脑钠肽(B-type natriuretic peptide, BNP)的最适表达模式。结果表明,相比于诱导型胞内表达、诱导型分泌表达、组成型分泌表达模式,BNP通过组成型胞内表达可达到最高的表达量。表达产物经过NI-NTA亲和层析,可得到纯化的BNP产量为235.5 mg/L,纯度可达90%。该研究构建了一套谷氨酸棒状杆菌中外源蛋白不同表达模式组合的快速检测方法,并成功实现了BNP的表达,这对以谷氨酸棒状杆菌作为宿主表达外源蛋白具有重要的指导意义。

本文引用格式

彭枫, 刘秀霞, 陈静, 等 . 不同表达模式组合对谷氨酸棒状杆菌中脑钠肽蛋白表达的影响[J]. 食品与发酵工业, 2018 , 44(8) : 1 -7 . DOI: 10.13995/j.cnki.11-1802/ts.016910

Abstract

For researching the expression model of B-type natriuretic peptide (BNP) in Corynebacterium glutamicum, a fast protein expression system was constructed using GFP as a model protein. The results showed that constitutive intracellular patterns were the best way for BNP expression compared with inducible intracellular, constitutive secretory and inducible secretory patterns. The BNP was purified and its production was about 235.5 mg/L with high purity (>90%). Conclusively, a fast protein expression system with different expression patterns was constructed in C. glutamicum, and BNP was successfully expressed using this system. This will greatly facilitate the application of this important platform organism.
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