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Title: Production of excreted human epidermal growth factor (hEGF) by an efficient recombinant Escherichia coli system
Authors: Sivakesava, S.
Xu, Z. N.
Chen, Y. H.
Hackett, James A.
Huang, R. C.
Lam, E.
Lam, T. L.
Siu, K. L.
Wong, R. S. C.
Wong, Wan-Keung R.
Keywords: hEGF
Escherichia coli
Human epidermal growth factor
Issue Date: Oct-1999
Citation: Process biochemistry, v. 34, no. 9, October 1999, p. 893-900
Abstract: Recombinant Escherichia coli JM101 strains harboring plasmids pWKW2 or lacUV5par8EGF, both encoding human epidermal growth factor (hEGF), were used in fermentations to optimize levels of excreted hEGF. Medium composition, inducer level, growth stage at induction, and culture conditions, were optimized with respect to volumetric production of the recombinant protein. MMBL medium, with glucose at 5g/l, and tryptone as nitrogen source, was chosen. Isopropyl-β-D-thiogalactopyranoside(IPTG) concentrations of 0.1mM for E. coli JM101 [pWKW2] and 0.2mM for E. coli K-12 JM101 [lacUV5par8EGF], were found to give the best hEGF production levels. The volumetric yields of hEGF were maximal when the cultures were induced in the mid-logarithmic phase. Growth temperature had a significant effect on hEGF yield. A simple continuous fed-batch process for cultivation of E. coli JM101[pWKW2] was developed. The maximum attained concentration of excreted hEGF in continuous fed-batch cultivation was 325mg/l, as compared to 175mg/l in batch cultivation. The hEGF produced from the continuous fed-batch cultivation was substantiated by SDS-PAGE and immunoblotting.
Rights: Process Biochemistry © copyright (1999) Elsevier. The Journal's web site is located at
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