Generation of the human metabolite piceatannol from the anticancer-preventive agent resveratrol by bacterial cytochrome P450 BM3.
In recent studies, the wild-type and mutant forms of cytochrome P450 (P450) BM3 (CYP102A1) from Bacillus megaterium were found to metabolize various drugs through reactions similar to those catalyzed by human P450 enzymes. Therefore, it was suggested that CYP102A1 can be used to produce large quantities of the metabolites of human P450-catalyzed reactions. trans-Resveratrol (3,4',5-trihydroxystilbene), an anticancer-preventive agent, is oxidized by human P450 1A2 to produce two major metabolites, piceatannol (3,5,3',4'-tetrahydroxystilbene) and another hydroxylated product. In this report, we show that the oxidation of trans-resveratrol, a human P450 1A2 substrate, is catalyzed by wild-type and a set of CYP102A1 mutants. One major hydroxylated product, piceatannol, was produced as a result of the hydroxylation reaction. Other hydroxylated products were not produced. Piceatannol formation was confirmed by high-performance liquid chromatography and gas chromatograph-mass spectrometry by comparing the metabolite with the authentic piceatannol compound. These results demonstrate that CYP102A1 mutants can be used to produce piceatannol, a human metabolite of resveratrol.
Kim
Dong-Hyun
DH
School of Biological Sciences and Technology, Chonnam National University, Gwangju 500-757, Republic of Korea.
Ahn
Taeho
T
Jung
Heung-Chae
HC
Pan
Jae-Gu
JG
Yun
Chul-Ho
CH
D016588:Anticarcinogenic Agents / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002248:Carbon Monoxide / Q000737:chemistry / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006900:Hydroxylation; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000502:physiology; D009251:NADPH-Ferrihemoprotein Reductase / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D000077185:Resveratrol; D013267:Stilbenes / Q000378:metabolism*
D016588:Anticarcinogenic Agents; D001426:Bacterial Proteins; D013267:Stilbenes; C041525:3,3',4,5'-tetrahydroxystilbene; D002248:Carbon Monoxide; D003577:Cytochrome P-450 Enzyme System; D009251:NADPH-Ferrihemoprotein Reductase; C052423:flavocytochrome P450 BM3 monoxygenases; D000077185:Resveratrol