IngentaConnect Enhanced Expression of Cytochrome P450s from lac-Based Plasmids U...

Authors: Woyski D.; Cupp-Vickery J.R.

Source: Archives of Biochemistry and Biophysics, Volume 388, Number 2, April 2001 , pp. 276-280(5)

Abstract:

The cytochrome P450 expression systems used in Escherichia coli are highly regulated and involve the use of the lac repressor to control expression. Induction in these systems utilizes the nonmetabolizable analog of lactose, isopropyl- beta -d-thiogalactopyranoside (IPTG), which is the most expensive compound required for an E. coli expression system. To determine if the natural inducer lactose could be used to induce cytochrome P450 expression we examined the expression of three P450 enzymes in E. coli using two different expression systems, pTrc99A and the T7-based PET22b vector. For both systems lactose was found to induce expression of active P450 to concentrations that exceeded the levels achieved with IPTG. A 20-liter fermentation of a P450 expression system in the pTrc plasmid in which lactose was used as the inducer resulted in 2.4 mol P450/liter, with a total yield of 2 g of cytochrome P450. The use of lactose for protein expression in E. coli should be broadly useful for the inexpensive, large-scale production of heterologous proteins in E. coli. Copyright 2001 Academic Press.

Keywords: cytochrome P450s; expression; lactose; IPTG

Language: English

Document Type: Research article

Affiliations: Department of Chemistry and Biochemistry, California State University, Fullerton, 800 N. State College Boulevard, Fullerton, California, 92834:

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Enhanced Expression of Cytochrome P450s from lac-Based Plasmids Using Lactose as the Inducer