重组大肠杆菌高密度发酵表达胰高血糖素样肽-1衍生物(GP62)的培养基优化(英文)  

High cell-density production of glucagon-like peptide-1 derivate(GP62) in recombinant E.coli by optimization of culture medium

作　　者：蒋德旗[1] 周天鸿[1] 冉艳红[1] 陈英[1] 桑延霞[1] 李弘剑[1] 

机构地区：[1]暨南大学生命科学技术学院生物工程学系,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2011年第3期334-338,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：the Ministry of Science and Technology of China(2009ZX09103-606);Guangzhou Scientific and Technology Commission(2009Z1-E151)

摘　　要：对重组E.coli BL21(DE3)高密度表达胰高血糖素样肽-1衍生物(GP62)的发酵培养基进行了优化.通过单因素实验与正交实验相结合的方法,依次对基础培养基,发酵培养基中碳源、氮源、无机盐等成份进行优化.在2YT培养基基础上,获得优化的发酵培养基2YT-GP62.在5L全自动发酵罐中利用终浓度0.5 mmol/L IPTG诱导,可获得的细胞密度为细胞干质量6.82 g/L发酵液(OD600=21.3),为LB培养基中的4.4倍,目的蛋白表达率由27.5%上升到31.2%,融合蛋白得率为4.40 mg/g干细胞.结果证明优化后的发酵培养基可用于重组GLP-1衍生物分批发酵生产.This study aims to get high expression level of GLP-1 derivate （GP62） in high cell-density recombinant Escherichia coli BI21 （ DE3 ） by optimizing its fed-batch culture medium. The effects of different basal medium, yeast extract and inorganic salt concentration on cell growth and expression of GLP-1 derivate （GP62） in recombinant E. coli BI21 （DE3） were investigated by single factor test and orthogonal array. Optimized 2YT-GP62 medium was obtained by improving the 2YT medium. The highest productivity of 4. 40 mg fusion protein per gram of dry cells and the final cell density of 21.3 （ OD600 ） were obtained in 5L fermentor induced by O. 5 mmol/L IPTG. The final cell density was 4. 4 times of that in LB culture me- dium and the expression level of GP62 was improved from 27.5% to 31.2%. The results show that 2YT- GP62 medium can be used for batch cultivation for production of recombinant GLP-1 derivate.

关 键 词：大肠杆菌 培养基 IPTG GLP-1 

分 类 号：Q939.9[生物学—微生物学]