Please use this identifier to cite or link to this item: http://hdl.handle.net/1783.1/2148

Production of excreted human epidermal growth factor (hEGF) by an efficient recombinant Escherichia coli system

Authors Sivakesava, S
Xu, ZN
Chen, YH
Hackett, J
Huang, RC
Lam, E
Lam, TL
Siu, KL
Wong, RSC
Wong, WKR
Issue Date 1999
Source Process biochemistry , v. 34, (9), 1999, OCT, p. 893-900
Summary Recombinant Escherichia coli JM101 strains harbouring 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 5 g/l and tryptone as nitrogen source, was chosen. Isopropyl-beta-D-thiogalactopyranoside(IPTG) concentrations of 0.1 mM for E. coli JM101[pWKW2] and 0.2 mM 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. call JM101[pWKW2] was developed. The maximum concentration of excreted hEGF attained in continuous fed-batch cultivation was 325 mg/l, as compared to 175 mg/l, in batch cultivation. The hEGF produced from the continuous fed-batch cultivation was substantiated by SDS-PAGE and immunoblotting. (C) 1999 Elsevier Science Ltd. All rights reserved.
Subjects
ISSN 0032-9592
Rights Process Biochemistry © copyright (1999) Elsevier. The Journal's web site is located at http://www.sciencedirect.com/
Language English
Format Article
Access View full-text via DOI
View full-text via Web of Science
View full-text via Scopus
Find@HKUST
Files in this item:
File Description Size Format
1_2prod.pdf 1894410 B Adobe PDF