PURIFICATION AND RECONSTITUTION OF THE ELECTRON-TRANSPORT COMPONENTS FOR 6-DEOXYERYTHRONOLIDE B-HYDROXYLASE, A CYTOCHROME-P-450 ENZYME OF MACROLIDE ANTIBIOTIC (ERYTHROMYCIN) BIOSYNTHESIS
PURIFICATION AND RECONSTITUTION OF THE ELECTRON-TRANSPORT COMPONENTS FOR 6-DEOXYERYTHRONOLIDE B-HYDROXYLASE, A CYTOCHROME-P-450 ENZYME OF MACROLIDE ANTIBIOTIC (ERYTHROMYCIN) BIOSYNTHESIS
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DOI：
10.1128/jb.170.4.1548-1553.1988
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发表时间：
1988-04-01
影响因子：
3.2
通讯作者：
HUTCHINSON, CR
中科院分区：
文献类型：
--
作者：
SHAFIEE, A;HUTCHINSON, CR
The hydroxylation of 6-deoxyerythronolide B (6D) to erythronolide B, a step in the biosynthesis of the 14-membered macrolide antibiotic erythromycin A by Saccharopolyspora erythraea, is catalyzed by a cytocyhrome P-450 monooxygenase that requires two electron transport proteins for the function of this terminal hydroxylase (A. Shafiee and C, R. Hutchinson, Biochemistry 26:6204-6210, 1987). Two flavoproteins and an iron-sulfur protein (erythrodoxin) were purified from S. erythaea CA340 and shown to act with 6D hydroxylase to catalyze the hydroxylation of (9R)-[9-3H]9-deoxo-9-hydroxy-6D in vitro in a sutiably reconstituted system. The flavoproteins contained flavin adenine dinucleotide and exhibited characteristic absorption maxima at 356 and 456 nm. The one with an Mr of 47,000 showed NADPH-dependent diaphorase and cytochrome c reductase activity, and the other, with an Mr of 53,000 showed NADH-dependent activities of the same two types. Erythrodoxin contained acid-labile sulfur and iron, had an Mr of 27,500, and showed a broad absorption maximum between 394 and 404 nm. The sequence of its first 15 amino acids, except for position 12, was the same as that the ferredoxin from Mycobacterium smegmatis.
