Purification of cytochrome P450 from filamentous fungus Rhyzopus nigricans

It has been reported in our previous studies that steroid hydroxylation sys tem of Rhizopus nigricans involves cytochrome P450 and NADPH-cytochrome P45 0 reductase in the electron transport system. Both enzymes are membrane bou nd and are located in the microsomal preparations of progesterone induced f ungal mycelia. In order to identify and characterize the cytochrome P450 co mponent of fungal monoxygenase system, microsomal proteins from induced myc elia were subjected to HIGH Q anion exchange and MONO P (FPLC) anion exchan ge chromatography. Four fractions containing cytochrome P450 have been reso lved on MONO P column. They exhibit CO difference spectra and type II diffe rence spectra with ketoconazole.