Secretory and extracellular production of recombinant proteins using Escherichia coli
- 7.6k Downloads
Escherichia coli is one of the most widely used hosts for the production of recombinant proteins. However, there are often problems in recovering substantial yields of correctly folded proteins. One approach to solve these problems is to have recombinant proteins secreted into the periplasmic space or culture medium. The secretory production of recombinant proteins has several advantages, such as simplicity of purification, avoidance of protease attack and N-terminal Met extension, and a better chance of correct protein folding. In addition to the well-established Sec system, the twin-arginine translocation (TAT) system has recently been employed for the efficient secretion of folded proteins. Various strategies for the extracellular production of recombinant proteins have also been developed. For the secretory production of complex proteins, periplasmic chaperones and protease can be manipulated to improve the yields of secreted proteins. This review discusses recent advances in secretory and extracellular production of recombinant proteins using E. coli.
KeywordsRecombinant Protein Disulfide Bond Secretory Production TMAO Periplasmic Space
This review was supported by the Korean Systems Biology Research Grant (M10309020000-03B5002-00000) from the Ministry of Science and Technology. Support from IBM through the IBM-SUR program is greatly appreciated.
- Arie J, Sassoon N, Betton J (2001) Chaperone function of FkpA, a heat shock prolyl isomerase, in the periplasm of Escherichia coli. Mol Microbiol 39:199–210Google Scholar
- Guisez Y, Fache I, Campfield LA, Smith FJ, Farid A, Plaetinck G, Van der Heyden J, Tavernier J, Fiers W, Burn P, Devos R (1998) Efficient secretion of biologically active recombinant OB protein (leptin) in Escherichia coli, purification from the periplasm and characterization. Protein Expr Purif 12:249–258CrossRefPubMedGoogle Scholar
- Kujau MJ, Hoischen C, Riesenberg D, Gumpert J (1998) Expression and secretion of functional miniantibodies McPC603scFvDhlx in cell-wall-less L-form strains of Proteus mirabilis and Escherichia coli: A comparison of the synthesis capacities of L-form strains with an E. coli producer strain. Appl Microbiol Biotechnol 49:51–58PubMedGoogle Scholar
- Nagahari K, Kanaya S, Munakata D, Aoyagi Y, Mizushima S (1985) Secretion into the culture medium of a foreign gene product from Escherichia coli: use of the ompF gene for secretion of human β-endorphin. EMBO J 16:3589–3592Google Scholar
- Pritchard MP, Ossetian R, Li DN, Henderson CJ, Burchell B, Wolf CR, Friedberg T (1997) A general strategy for the expression of recombinant human cytochrome P450 s in Escherichia coli using bacterial signal peptides: Expression of CYP3A4, CYP2A6, and CYP2E1. Arch Biochem Biophys 345:342–354CrossRefPubMedGoogle Scholar
- Rippmann JF, Klein M, Hoischen C, Brocks B, Rettig WJ, Gumpert J, Pfizenmaier K, Mattes R, Moosmayer D (1998) Procaryotic expression of single-chain variable-fragment (scFv) antibodies: secretion in L-form cells of Proteus mirabilis leads to active product and overcomes the limitations of periplasmic expression in Escherichia coli. Appl Environ Microbiol 64(12):4862-4869.PubMedGoogle Scholar
- Shokri A, Saden AM and Larsson G (2003) Cell and process design for targeting of recombinant protein into the culture medium of Escherichia coli. Appl Microbial Biotechnol 60:654–664Google Scholar
- Tanaka T, Horio T, Matuo Y (2002) Secretory production of recombinant human C-reactive protein in Escherichia coli, capable of binding with phosphorylcholine, and its characterization. Biochem Biophys Res Commun, 295(1):163–6Google Scholar
- Uchida H, Naito N, Asada N, Wada M, Ikeda M, Kobayashi H, Asanagi M, Mori K, Fujita Y, Konda K, Kusuhara N, Kamioka T, Nakashima K, Honjo M (1997) Secretion of authentic 20-kDa human growth hormone (20 K hGH) in Escherichia coli and properties of the purified product. J Biotechnol 55:101–112CrossRefPubMedGoogle Scholar
- Van der Wal FJ, Koningstein G, ten Hagen CM, Oudega B, Luirink J (1998) Optimization of bacterocin release protein (BRP)-mediated protein release by Escherichia coli: random mutagenesis of the pCloDF13-Derived BRP gene to uncouple lethality and quasi-lysis from protein release. Appl Environ Microbiol 64:392–398PubMedGoogle Scholar
- Yang J, Moyana T, Mackenzie S, Xia Q and Xiang J (1998) One hundred Seventy-fold increase in excretion of an FV frangmant tumor necrosis factor alpha fusion protein (SFV/TNF-α) from Escherichia coli caused by the synergistic effects of glycine and triton X-100. Appl Environ Microbiol 64:2669–2874Google Scholar