Purification and characterization of a soybean flour-induced cytochrome P-450 from Streptomyces griseus.
A soybean flour-induced, soluble cytochrome P-450 (P-450soy) was purified 130-fold to homogeneity from Streptomyces griseus. Native cytochrome P-450soy is a single polypeptide, with a molecular weight of 47,500, in association with one ferriprotoporphyrin IX prosthetic group. Oxidized P-450soy exhibited visible absorption maxima at 394, 514, and 646 nm, characteristic of a high-spin cytochrome P-450. The CO-reduced difference spectrum of P-450soy had a Soret maximum at 448 nm. When reconstituted with spinach ferredoxin and spinach ferredoxin:NADP+ oxidoreductase, purified cytochrome P-450soy catalyzed the NADPH-dependent oxidation of the xenobiotic substrates precocene II and 7-ethoxycoumarin. In vitro proteolysis of cytochrome P-450soy generated a stable and catalytically active cytochrome P-450, designated P-450soy delta.
Trower
M K
MK
E. I. du Pont de Nemours and Company, Central Research and Development Department, Wilmington, Delaware 19880-0228.
Sariaslani
F S
FS
O'Keefe
D P
DP
D000596:Amino Acids / Q000032:analysis; D001711:Biotransformation; D015153:Blotting, Western; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification* / Q000378:metabolism; D004790:Enzyme Induction; D006418:Heme / Q000032:analysis; D007526:Isoelectric Point; D008970:Molecular Weight; D010449:Peptide Mapping; D013025:Glycine max; D013057:Spectrum Analysis; D013305:Streptomyces griseus / Q000201:enzymology*; D015262:Xenobiotics / Q000378:metabolism
D000596:Amino Acids; D015262:Xenobiotics; D006418:Heme; D003577:Cytochrome P-450 Enzyme System