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

Jung Min Song; Young Jun An; Mee Hye Kang; Youn Ho Lee; Sun Shin Cha

doi:10.1016/j.pep.2012.01.020

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

Jung Min Song, Young Jun An, Mee Hye Kang, Youn Ho Lee, Sun Shin Cha

Department of Chemistry & Nanoscience

Research output: Contribution to journal › Article › peer-review

52 Scopus citations

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 0.9 mg of Cryptopygus antarticus mannanase (CaMan) and cellulase (CaCel) from 1 l 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.

Original language	English
Pages (from-to)	297-301
Number of pages	5
Journal	Protein Expression and Purification
Volume	82
Issue number	2
DOIs	https://doi.org/10.1016/j.pep.2012.01.020
State	Published - Apr 2012

Bibliographical note

Funding Information:
We are grateful to Mr. Jung Ho Jeon of Korea Ocean Research and Development Institute (Korea) for contributing the pET-EM3L7 expression vector. This work was supported by the grant ( PP00740 ) from the Korea Ocean Research and Development Institute and by the Marine & Extreme Genome Research Center Program from the Ministry of Land, Transport, and Maritime Affairs.

Keywords

E. coli
Extremely low temperature
Protein expression

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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 0.9 mg of Cryptopygus antarticus mannanase (CaMan) and cellulase (CaCel) from 1 l 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.",

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note = "Funding Information: We are grateful to Mr. Jung Ho Jeon of Korea Ocean Research and Development Institute (Korea) for contributing the pET-EM3L7 expression vector. This work was supported by the grant ( PP00740 ) from the Korea Ocean Research and Development Institute and by the Marine & Extreme Genome Research Center Program from the Ministry of Land, Transport, and Maritime Affairs. ",

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AU - Lee, Youn Ho

AU - Cha, Sun Shin

N1 - Funding Information: We are grateful to Mr. Jung Ho Jeon of Korea Ocean Research and Development Institute (Korea) for contributing the pET-EM3L7 expression vector. This work was supported by the grant ( PP00740 ) from the Korea Ocean Research and Development Institute and by the Marine & Extreme Genome Research Center Program from the Ministry of Land, Transport, and Maritime Affairs.

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N2 - 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 0.9 mg of Cryptopygus antarticus mannanase (CaMan) and cellulase (CaCel) from 1 l 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.

AB - 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 0.9 mg of Cryptopygus antarticus mannanase (CaMan) and cellulase (CaCel) from 1 l 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.

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

Abstract

Bibliographical note

Keywords

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