Recombinant gene expression in Escherichia coli cultivation using lactose as inducer

• Published in: Journal of biotechnology 1998-02, Vol.60 (1-2), p.47-54
• Main Authors: Gombert, A.K, Kilikian, B.V
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Biotechnology; Chickens; DNA-Directed RNA Polymerases - metabolism; Escherichia coli - genetics; Escherichia coli - growth & development; Fermentation; Fundamental and applied biological sciences. Psychology; Galactose - metabolism; Gene Expression - drug effects; Induction; Lac UV5; Lactose; Lactose - pharmacology; Methods. Procedures. Technologies; Microbial engineering. Fermentation and microbial culture technology; Muscles - chemistry; Promoter Regions, Genetic; Recombinant Escherichia coli; Recombinant Proteins; Troponin - genetics; Troponin C; Troponin T; Viral Proteins
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/S0168-1656(97)00185-5

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 l−1 dry cell weight (DCW)—before induction started—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 g l−1) together with the second and the third lactose pulses. The results presented suggest that the residual lactose concentration plays an important role on the production of the heterologous protein.