Recombinant gene expression in Escherichia coli cultivation using lactose as inducer

被引:61
作者
Gombert, AK [1 ]
Kilikian, BV [1 ]
机构
[1] Univ Sao Paulo, Escola Politecn, Dept Engn Quim, BR-05424970 Sao Paulo, SP, Brazil
基金
巴西圣保罗研究基金会;
关键词
recombinant Escherichia coli; lac UV5; induction; lactose; troponin C;
D O I
10.1016/S0168-1656(97)00185-5
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The use of lactose as inducer of foreign gene expression under control of the lac UV5 promoter was investigated in recombinant Escherichia coli. Chicken muscle troponin C (TnC) gene was transcripted by T7 RNA polymerase and expressed in bioreactor cultivations after a feed-forward controlled fed-batch growth phase. Cell concentrations of 22 g(-1) dry cell weight (DCW)-before induction starred-were used to achieve a TnC content of 19.5% of total cell protein through an induction strategy that combined the addition of a specific lactose amount of 4.7 g g(-1) DCW divided into three pulses and the addition of yeast extract (1 g1(-1)) together with the second and the third lactose pulses. The results presented suggest that the residual lactose concentration play:; an important role on the production of the heterologous protein. (C) 1998 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:47 / 54
页数:8
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