Tricistronic overexpression of cytochrome P450cam, putidaredoxin, and putidaredoxin reductase provides a useful cell-based catalytic system.
The catalytic turnover of cytochrome P450( cam ) from Pseudomonas putida requires two auxiliary reduction partners, putidaredoxin (Pd) and putidaredoxin reductase (PdR). We report the functional expression in Escherichia coli of tricistronic constructs consisting of P450( cam ) encoded by the first cistron and the auxiliary proteins, Pd and PdR by the second and the third. Transformed bacterial whole cells efficiently oxidized (1R)-(+)-camphor to 5-exo-hydroxycamphor and, interestingly, limonene to (-)-perillyl alcohol. These bioengineered E. coli cells possess a heterologous self-sufficient P450 catalytic system that may have advantages in terms of low cost and high yield for the production of fine chemicals.
Kim
Donghak
D
Department of Biological Sciences, Konkuk University, 1 Hwayang-dong, Gwangjn-gu, Seoul 143-701, Korea. donghak@konkuk.ac.kr
Ortiz de Montellano
Paul R
PR
D001711:Biotransformation; D002164:Camphor / Q000378:metabolism; D019475:Camphor 5-Monooxygenase / Q000096:biosynthesis* / Q000235:genetics; D053138:Cyclohexenes / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005288:Ferredoxins / Q000096:biosynthesis* / Q000235:genetics; D018628:Gene Dosage; D015870:Gene Expression; D000077222:Limonene; D039821:Monoterpenes / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics; D016958:Pseudomonas putida / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013729:Terpenes / Q000378:metabolism
D053138:Cyclohexenes; D005288:Ferredoxins; D039821:Monoterpenes; D011994:Recombinant Proteins; D013729:Terpenes; C032208:perillyl alcohol; C100137:putidaredoxin; D002164:Camphor; D000077222:Limonene; D019475:Camphor 5-Monooxygenase; D009247:NADH, NADPH Oxidoreductases; C022486:putidaredoxin reductase