CLONING, CHARACTERIZATION AND EXPRESSION OF THE GENE ENCODING CYTOCHROME P-450(SCA-2) FROM STREPTOMYCES-CARBOPHILUS INVOLVED IN PRODUCTION OF PRAVASTATIN, A SPECIFIC HMG-COA REDUCTASE INHIBITOR

Pravastatin, a drug for treating hypercholesterolemia, is produced by hydroxylation of ML-236B . Na in Streptomyces carbophilus (Sc) catalyz ed by the cytochrome P-450(sca) (CytP-450(sca)) monooxygenase system. The gene (cytP-450(sca-2)) encoding CytP-450(sca-2) was cloned from Sc . The gene had an open reading frame of 1233 bp, encoding a 410-amino- acid protein. The partial sequencing of the purified CytP-450(sca-2) r evealed that the N-terminal Met had been removed. CytP-450(sca-2) cont ained the heme-binding HR2 region characteristic of all CytP-450, as w ell as the putative oxygen-binding site proposed in CytP-450(cam) from Pseudomonas putida. ML-236B . Na enhanced transcription of cytP-450(s ca-2), suggesting that substrate induction in Sc is transcriptionally regulated. S. lividans (Sl) transfected with cytP-450(sca-2) converted ML-236B . Na to pravastatin, indicating the cloned gene to be functio nal in Sl.