Cultivation at 6-10 掳C is an effective strategy to overcome the insolubility of recombinant proteins in Escherichia coli

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• 作者：Jung Min Song^a ; Young Jun An^b ; Mee Hye Kang^a ; Youn-Ho Lee^a ; ^{ylee@kordi.re.kr} ; Sun-Shin Cha^b ; ^{chajung@kordi.re.kr}
• 关键词：Protein expression ; E. coli ; Extremely low temperature
• 刊名：Protein Expression and Purification
• 出版年：2012
• 期刊代码：184_10465928
• 类别：bio
• 出版时间：April, 2012
• 卷：82
• 期：2
• 页码：297-301
• 文件大小：492 K

摘要

Protein expression in Escherichia coli at 15-25 掳C is widely used to increase the solubility of recombinant proteins. However, many recombinant proteins are insolubly expressed even at those low temperatures. Here, we show that recombinant proteins can be expressed as soluble forms by simply lowering temperature to 6-10 掳C without cold adapted chaperon systems. By using E. coli Rosetta-gami2(DE3), we obtained 1.8 and 0.9 mg of Cryptopygus antarticus mannanase (CaMan) and cellulase (CaCel) from 1 l culture grown at 6 and 10 掳C, respectively. Cultivation at 10 掳C also led to successful expression of EM3L7 (a lipase isolated from a metagenomic library) in a soluble form in E. coli BL21(DE3). Consequently, E. coli cultivation at 6-10 掳C is an effective strategy for overcoming a major hurdle of the inclusion body formation.