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Comparison of Two Vectors for Functional Expression of a Bacterial Cytochrome P450 Gene in Escherichia coli Using CYP153 Genes

2009; Oxford University Press; Volume: 73; Issue: 8 Linguagem: Inglês

10.1271/bbb.90199

1347-6947

Naoya FUJITA, Futoshi Sumisa, Kazutoshi Shindo, Hiroki Kabumoto, Akira Arisawa, Hiroshi Ikenaga, Norihiko Misawa,

Computational Drug Discovery Methods

Two vectors, pT7NScamAB and pRED, have been used for the functional expression of bacterial class I cytochrome P450 (P450) genes in Escherichia coli, which utilize putidaredoxin reductase (CamA) and putidaredoxin (CamB), and the reductase domain of a self-sufficient P450RhF respectively, for electron transfer from NAD(P)H to a P450 protein. We here compared the efficiency of bioconversion with the two vectors towards n-octane, cyclohexane, n-butylbenzene, and 2-n-butylbenzofuran using two well-characterized CYP153A genes, aciA and CYP153A13a (P450balk). As for n-octane bioconversion, aciA and pT7camAB was the best combination for the production of 1-octanol and 1,8-octanediol. As for the bioconversion of cyclohexane, n-butylbenzene and 2-n-butylbenzofuran, CYP153A13a with pRED achieved the most efficient bioconversion, as compared by conversion ratio per active CYP153A protein content. It was also found that 2-n-butylbenzofuran is biotransformed into 4-benzofuran-2-yl-butyric acid via 4-benzofuran-2-yl-butan-1-ol with E. coli cells expressing CYP153A.