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Strategies for the Production of Recombinant Protein in Escherichia coli

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Abstract

In the recent past years, a large number of proteins have been expressed in Escherichia coli with high productivity due to rapid development of genetic engineering technologies. There are many hosts used for the production of recombinant protein but the preferred choice is E. coli due to its easier culture, short life cycle, well-known genetics, and easy genetic manipulation. We often face a problem in the expression of foreign genes in E. coli. Soluble recombinant protein is a prerequisite for structural, functional and biochemical studies of a protein. Researchers often face problems producing soluble recombinant proteins for over-expression, mainly the expression and solubility of heterologous proteins. There is no universal strategy to solve these problems but there are a few methods that can improve the level of expression, non-expression, or less expression of the gene of interest in E. coli. This review addresses these issues properly. Five levels of strategies can be used to increase the expression and solubility of over-expressed protein; (1) changing the vector, (2) changing the host, (3) changing the culture parameters of the recombinant host strain, (4) co-expression of other genes and (5) changing the gene sequences, which may help increase expression and the proper folding of desired protein. Here we present the resources available for the expression of a gene in E. coli to get a substantial amount of good quality recombinant protein. The resources include different strains of E. coli, different E. coli expression vectors, different physical and chemical agents and the co expression of chaperone interacting proteins. Perhaps it would be the solutions to such problems that will finally lead to the maturity of the application of recombinant proteins. The proposed solutions to such problems will finally lead to the maturity of the application of recombinant proteins.

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Abbreviations

aa:

Amino acid

bp:

Base pairs

GST:

Glutathione S-transferase

HAT:

Histone acetyltransferase

IPTG:

Isopropylthio-β-galactoside

MBP:

Maltose binding protein

NEB:

New England Biolabs

MCS:

Multiple cloning site

Ni-NTA:

Nickel nitrilotriacetic acid

ORF:

Open reading frame

SUMO:

Small ubiquitin modifier

Trx:

Thioredoxin

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Acknowledgments

G.J.G. acknowledges the University Grant Commission (UGC), New Delhi, India for endowing him the Dr. D. S. Kothari Post Doctoral Fellowship and Prof. Rakesh Bhatnagar [School of Biotechnology, Jawaharlal Nehru University (JNU), New Delhi, India] for providing the opportunity to work in his lab. The authors are thankful to the Departments of Biotechnology, JNU, New Delhi, India and National Institute of Technology, Raipur (CG), India.

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Authors and Affiliations

  1. School of Biotechnology, Jawaharlal Nehru University, New Delhi, 110067, India

    Gopal Jee Gopal

  2. Department of Biotechnology, National Institute of Technology, Raipur, 492010, Chhattisgarh, India

    Awanish Kumar

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  1. Gopal Jee Gopal
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  2. Awanish Kumar
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Correspondence to Awanish Kumar.

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Gopal, G.J., Kumar, A. Strategies for the Production of Recombinant Protein in Escherichia coli . Protein J 32, 419–425 (2013). https://doi.org/10.1007/s10930-013-9502-5

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