title: Cultivation at 6-10°C is an effective strategy to overcome the insolubility of recombinant proteins in Escherichia coli.
abstract: Protein expression in Escherichia coli at 15-25°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°C without cold adapted chaperon systems. By using E. coli Rosetta-gami2(DE3), we obtained 1.8 and 0.9mg of Cryptopygus antarticus mannanase (CaMan) and cellulase (CaCel) from 1l culture grown at 6 and 10°C, respectively. Cultivation at 10°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°C is an effective strategy for overcoming a major hurdle of the inclusion body formation.
authors.affiliation: Marine Biology and Living Resources Research Department, Korea Ocean Research and Development Institute, Ansan 426-744, Republic of Korea.
authors.forename: Jung Min
authors.initials: JM
authors.lastname: Song
authors.forename: Young Jun
authors.initials: YJ
authors.lastname: An
authors.forename: Mee Hye
authors.initials: MH
authors.lastname: Kang
authors.forename: Youn-Ho
authors.initials: YH
authors.lastname: Lee
authors.forename: Sun-Shin
authors.initials: SS
authors.lastname: Cha
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry; D002480:Cellulase / Q000096:biosynthesis* / Q000737:chemistry; D003080:Cold Temperature; D046508:Culture Techniques; D004926:Escherichia coli / Q000254:growth & development* / Q000378:metabolism; D019476:Insect Proteins / Q000096:biosynthesis* / Q000737:chemistry; D008049:Lipase / Q000096:biosynthesis* / Q000737:chemistry; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D012995:Solubility
chemical_list: D001426:Bacterial Proteins; D019476:Insect Proteins; D011994:Recombinant Proteins; D008049:Lipase; D002480:Cellulase