摘要
小分子伴侣GroEL(191-345)是分子伴侣GroEL顶端区域氨基酸残基191-345的片断,它能显著提高基因工程蛋白蝎毒Cn5、亲环蛋白A和吲哚3-甘油磷酸合成酶等的复性效率,具有广阔的应用前景。为制得大量的小分子伴侣进行蛋白质复性研究,本文对其培养条件进行了优化。结果表明携带小分子伴侣基因的工程菌在含有无机盐和碳源的M9培养基中培养能够显著提高小分子伴侣的表达量,在确定M9为基本培养基之后,对其主要成分进行了优化;同时,考察了发酵温度、供氧量、诱导条件对表达量的影响,将发酵产量提高到556.3mg/L。
The apical domain of GroEL (residues 191-345) is expressed in E.coli to give a functional mini-chaperone, and the refolding yields of scorpion toxin Cn5, cyclophilin A and IGPS assisted by mini-chaperone were improved remarkably. Owing to its potentially broad application, the optimum culture condition of mini-chaperone was studied in detail. M9 medium with carbon source and inorganic salts was found to be suitable for the expression of mini-chaperone GroEL (191-345) in E. coli. M9 medium was then reformulated and the optimum temperature, oxygen demand and induction conditions were carefully chosen to improve the mini-chaperone production to 556.3 mg/L.
基金
浙江省自然科学基金资助项目(201099)
国家教育部留学回国人员科研启动基金资助项目.
