Comparison of two vectors for functional expression of a bacterial cytochrome P450 gene in Escherichia coli using CYP153 genes.
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.
Fujita
Naoya
N
Marine Biotechnology Institute, Heita, Kamaishi, Japan.
Sumisa
Futoshi
F
Shindo
Kazutoshi
K
Kabumoto
Hiroki
H
Arisawa
Akira
A
Ikenaga
Hiroshi
H
Misawa
Norihiko
N
D000595:Amino Acid Sequence; D001572:Benzofurans / Q000378:metabolism; D002470:Cell Survival; D003510:Cyclohexanes / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D009795:Octanes / Q000378:metabolism
D001572:Benzofurans; D003510:Cyclohexanes; D009795:Octanes; C506365:Cyclohexane; D003577:Cytochrome P-450 Enzyme System; C435228:cytochrome P-450 CYP153 (Acinetobacter); C105430:benzofuran; C026728:octane