A two component-type cytochrome P-450 monooxygenase system in a prokaryote that catalyzes hydroxylation of ML-236B to pravastatin, a tissue-selective inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase.
Pravastatin (CS-514) is a tissue selective inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase (EC 1.1.1.34), a key enzyme in cholesterol biosynthesis. This compound is obtained by hydroxylation of ML-236B (mevastatin) in Streptomyces carbophilus catalyzed by a cytochrome P-450sca monooxygenase system. NADH-cytochrome P-450 reductase was purified to homogeneity from S. carbophilus as a single polypeptide chain with a molecular weight of 51 kDa, and reconstituted the hydroxylation in vitro with cytochrome P-450sca, NADH and O2. This protein contained FAD and FMN molecule. The FMN molecule was easily dissociated from the reductase, and had a Kd value of 5 x 10(-5) M. The cytochrome P-450sca monooxygenase system was present in the soluble fraction and consisted of only two components, cytochrome P-450sca and flavoprotein. Our results constitute the demonstration of a two component-type cytochrome P-450 system in a prokaryote.
Serizawa
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N
Fermentation Research Laboratories, Sankyo Co., Ltd., Tokyo, Japan.
Matsuoka
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T
D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification* / Q000378:metabolism; D004795:Enzyme Stability; D005420:Flavoproteins / Q000302:isolation & purification; D006538:Heptanoic Acids / Q000378:metabolism*; D006900:Hydroxylation; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D008148:Lovastatin / Q000031:analogs & derivatives* / Q000494:pharmacology; D006899:Mixed Function Oxygenases / Q000302:isolation & purification; D008954:Models, Biological; D009247:NADH, NADPH Oxidoreductases / Q000302:isolation & purification; D009281:Naphthalenes / Q000378:metabolism*; D009711:Nucleotides / Q000494:pharmacology; D010105:Oxygenases / Q000302:isolation & purification* / Q000378:metabolism; D017035:Pravastatin; D013302:Streptomyces / Q000201:enzymology*
D005420:Flavoproteins; D006538:Heptanoic Acids; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D009281:Naphthalenes; D009711:Nucleotides; C012258:mevastatin; D003577:Cytochrome P-450 Enzyme System; D008148:Lovastatin; D006899:Mixed Function Oxygenases; D010105:Oxygenases; C069505:mevastatin hydroxylase; D009247:NADH, NADPH Oxidoreductases; C069506:NADH-cytochrome P-450 reductase; D017035:Pravastatin