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Mass production of human epidermal growth factor using fed‐batch cultures of recombinant Escherichia coli
Author(s) -
Shimizu Norio,
Fukuzono Shinichi,
Harada Yoshinori,
Fujimori Kiyoshi,
Gotoh Kenichi,
Yamazaki Yoshio
Publication year - 1991
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260380106
Subject(s) - escherichia coli , plasmid , recombinant dna , microbiology and biotechnology , gene , biology , inclusion bodies , enterobacteriaceae , gene expression , cell culture , epidermal growth factor , chemistry , biochemistry , genetics
Fed‐batch cultures of recombinant E. coli HB101 harboring expression plasmid pTRLBT1 or pTREBT1, with acetate concentration monitoring, are investigated to obtain high cell density and large amounts of human epidermal growth factor (hEGF). The expression plasmid pTRlBT1 contains a synthetic hEGF gene attached downstream of the N‐terminal fragment of the trp L gene preceded by the trp promoter. The expression plasmid pTREBT1 contains the same coding sequence attached downstream of the N‐terminal fragment of the trp E gene preceded by the trp promoter, trp L gene, and attenuator region. E. coli harboring pTREBT1 produces 0.56 mg/L hEGE and immediately degrades it. On the other hand E. coli harboring pTRLBT1 produces 6.8 mg/L hEGF and does not decompose it. Prominent inclusion bodies are observed in E. coli cells harboring pTRLBT1 using an election microscope. To Cultivate E. coli harboring pTRLBT1, a fed‐batch culture system, divided into a cell growth step and an hEGF production step, is carried out. The cells grow smoothly without acetate‐induced inhibition. Cell concentration and hEGF quantity reach the high values of 21 g/L and 60 mg/L, respectively.