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Title 

Functional expression in Bacillus subtilis of mammalian NADPH-cytochrome P450 oxidoreductase and its spore-display

Authors 

Sung-Kun YimHeung Chae JungC H YunJae Gu Pan

Publisher 

Elsevier

Issue Date 

2009

Citation 

Protein Expression & Purification, vol. 63, no. 1, pp. 5-11

Keywords 

bacillus expressionCPRNADPH-cytochrome P450 reductasespore-displaywhole-cell biocatalystcytochrome ccytochrome P450 1A2bacillus subtilisbacterial sporegene expression

Abstract 

The technology for over-expressing NADPH-cytochrome P450 reductase (CPR), a diflavin-containing enzyme, offers the opportunity to develop enzymatic systems for environmental detoxication and bioconversions of drugs, pesticides and fine chemicals. In this study, Bacillus subtilis was chosen to express rat CPR (rCPR) because of its capacities for high protein production and spore formation. rCPR was expressed in B. subtilis DB104 under the transcriptional control of an IPTG-inducible fusion promoter of PgroE and Ptac. The expressed rCPR was released into the culture medium after sporulation by autolysis of the host cell. It was associated with and displayed on the spore surfaces; this was confirmed by measuring rCPR activity in purified spores and analyzing its accessibility to anti-rCPR antibodies using flow cytometry. The spore-displayed rCPR was able to reduce cytochrome c and ferricyanide, and also assisted in the O-deethylation of 7-ethoxyresorufin and 7-ethoxy-4-trifluoromethylcoumarin (EFC) by human cytochrome P450 1A2, indicating that it was functionally active. Spore surface display of rCPR in B. subtilis appears to be useful for preparing cytochrome P450-related enzymes, and spore biocatalysts of rCPR are likely to have wide biotechnological applications.

ISSN 

1046-5928

Link 

http://dx.doi.org/10.1016/j.pep.2008.07.004

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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