Abstract
β-Galactosidase and streptokinase expression was tested under the control of the T7 promoter in batch and fed-batch cultures. An Escherichia coli host GJ1158, which contained the T7 RNA polymerase gene under the osmo-responsive proUp promoter, was used for expression studies. β-Galactosidase expression was enhanced from 26 mg l−1 to 127 mg l−1 in batch culture when a combination of sucrose and sorbitol was used instead of salt as an inducer. Similarly in fed-batch cultures 140 mg streptokinase l−1 was formed with sucrose and sorbitol induction which was higher than that achieved with IPTG induced cultures.
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References
Bhandari P, Gowrishankar J (1997) An E. coli host strain useful for efficient overproduction of cloned gene products with NaCl as the inducer. J. Bacteriol. 179: 4403–4406.
Gowrishankar J, Manna D (1996) How is osmotic regulation of transcription of the E. coli proU operon achieved? A review and a model. Genetica 97: 363–378.
Grossman TH, Kawasaki ES, Punreddy SR, Osburne MS (1998) Spontaneous cAMP-dependent derepression of gene expression in stationary phase plays a role in recombinant expression instability. Gene 209: 95–103.
Gupta JC, Jaisani M, Pandey G, Mukherjee KJ (1999) Enhancing recombinant protein yields in E. coli using the T7 system under the control of heat inducible γP L promoter. J. Biotechnol. 68: 125–134.
Imanaka T, Aiba S (1981) A perspective on the application of genetic engineering stability of recombinant plasmid. Ann. N.Y. Acad. Sci. 369: 1–14.
Kim JY, Ryu DDY (1991) The effects of plasmid content, transcription efficiency on the productivity of a cloned gene protein in E. coli. Biotechnol. Bioeng. 38: 1271–1279.
Miller JH (1972) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press, pp. 352–355.
Studier W, Moffait BA (1986) Use of bacteriophage T7 RNA polymerase to direct selective high level expression of specific genes. J. Mol. Biol. 189: 115–130.
Studier FW, Rosenberg AH, Dunn JJ, Dubendorff JW (1990) Use of T7 polymerase to direct expression of cloned genes. Meth. Enzymol. 185: 60–89.
Tabor S, Richardson CC (1985) A bacteriophage T7 RNA polymerase/ promoter system for controlled exclusive expression of specific genes. Proc. Natl. Acad. Sci. USA 82: 1074–1078.
Yazdani SS, Mukherjee KJ (1998) Overexpression of streptokinase using a fed-batch strategy. Biotechnol. Lett. 20: 923–927.
Yoon SK, Kang WK, Park TH (1994) Fed-batch operation of recombinant E. coli containing trp promoter with controlled specific growth rate. Biotechnol. Bioeng. 43: 995–999.
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Pal, Y., Gupta, J.C. & Mukherjee, K. Optimizing recombinant protein expression in the T7 system under the control of the proUp promoter. Biotechnology Letters 23, 41–46 (2001). https://doi.org/10.1023/A:1026712310154
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DOI: https://doi.org/10.1023/A:1026712310154
- β-galactosidase
- proUp promoter
- streptokinase
- T7 promoter