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Protein Expr Purif. 2012 May;83(1):30-6. doi: 10.1016/j.pep.2012.02.014. Epub 2012 Mar 8.

Galactose can be an inducer for production of therapeutic proteins by auto-induction using E. coli BL21 strains.

Author information

1
Biologics Process Science, Technical Operations, Bristol-Myers Squibb, Syracuse, NY 13057, United States.

Abstract

Recently lactose mediated auto-induction in Escherichia coli has gained a lot of interest because higher protein titer could be achieved without the need to monitor growth and add inducer at the proper time. In this study a high level therapeutic protein production by auto-induction was observed in E. coli BL21 using either T7 or tac promoters in the modified Luria Bertani (mLB) medium containing soy peptone instead of tryptone in Luria Bertani (LB) medium. Based on medium analysis and spiking experiments it was found that 0.4 mM galactose from the soy peptone caused the auto-induction. E. coli cultures induced by galactose can saturate at considerably higher density than cultures induced by IPTG. Galactose is not consumed by E. coli BL21. Finally it has been demonstrated that auto-induction can be effectively used in fed-batch fermentation for the industrial production of a therapeutic protein. The principle of galactose mediated auto-induction should be able to apply to high throughput microplates, shake flasks and fed-batch fermentors for clone screening and therapeutic protein expression in E. coli gal(-) strains such as most commonly used BL21.

PMID:
22425658
DOI:
10.1016/j.pep.2012.02.014
[Indexed for MEDLINE]

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