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Appl Environ Microbiol. 2007 Nov;73(22):7246-51. Epub 2007 Sep 28.

Engineering of artificial plant cytochrome P450 enzymes for synthesis of isoflavones by Escherichia coli.

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  • 1Department of Chemical and Biological Engineering, The State University of New York, Buffalo, NY 14260, USA.

Abstract

Engineered microbes are becoming increasingly important as recombinant production platforms. However, the nonfunctionality of membrane-bound cytochrome P450 enzymes precludes the use of industrially relevant prokaryotes such as Escherichia coli for high-level in vivo synthesis of many functional plant-derived compounds. We describe the design of a series of artificial isoflavone synthases that allowed the robust production of plant estrogen pharmaceuticals by E. coli. Through this methodology, a plant P450 construct was assembled to mimic the architecture of a self-sufficient bacterial P450 and contained tailor-made membrane recognition signals. The specific in vivo production catalyzed by one identified chimera was up to 20-fold higher than that achieved by the native enzyme expressed in a eukaryotic host and up to 10-fold higher than production by plants. This novel biological device is a strategy for the utilization of laboratory bacteria to robustly manufacture high-value plant P450 products.

PMID:
17905887
[PubMed - indexed for MEDLINE]
PMCID:
PMC2168208
Free PMC Article
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