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Received May 11, 2009
Accepted September 6, 2009
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Biotransformation of medium-chain alkanes using recombinant P450 monooxygenase from Alcanivorax borkumensis SK2 expressed in Escherichia coli
Department of Food Science and Human Nutrition, and Research Institute of Human Ecology, Chonbuk National University, Jeonju, Jeonbuk 561-756, Korea
baiksh@chonbuk.ac.kr
Korean Journal of Chemical Engineering, March 2010, 27(3), 905-909(5), 10.1007/s11814-010-0131-9
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Abstract
A bioconversion system for medium-chain alkanes was constructed by using a recombinant Escherichia coli whole-cell biocatalyst expressing P450 monooxygenase genes, ferredoxin, and ferredoxin reductase cloned from Alcanivorax borkumensis as an operon. The recombinant E. coli harboring the P450 gene and two related expression component enzymes, ferredoxin and ferredoxin reductase, was constructed in a single vector pET21(a) and successfully expressed in E. coli BL21(DE3) as a soluble form, showing a molecular weight of 53 kDa on 10% SDS-PAGE. When the cell-free extract of E. coli BL21 expressing p450 monooxygenase was subjected to reduced CO difference spectral analysis, a soret band near 450 nm appeared indicating that the cloned P450 was expressed as a functionally active enzyme. The E. coli cells harboring the expressed P450 gene were able to convert n-octane and 1-decene, producing approximately 450 μg/ml of n-octanol and 290 μg/ml of 1,2-epoxydecane, respectively, at pH 7.0 and 30 ℃. However, the recombinant E. coli cells were not able to convert the branched alkane, 2,6,10,14-tetramethylpentadecane (C19).
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