Purification and characterization of cytochrome P450RR1 from Rhodococcus rhodochrous

摘要: A soluble cytochrome P450 whose synthesis is induced by and that binds 2-ethoxyphenol was purified to apparent homogeneity from Rhodococcus rhodochrous strain 116. The enzyme had a subunit molecular mass of 44.5 kDa as determined SDS/PAGE pI 5.2. electronic absorption spectrum indicates the native in absence substrate predominantly low-spin state (13% high-spin 50 mM Mops, pH 7.0 25 degrees C). 2-Methoxyphenol with macroscopic dissociation constant 0.53 +/- 0.03 microM (50 7.0, C) induces 99.7% transition heme iron pentacoordinate form. Using reconstituted in-vitro activity assay, it demonstrated P450RR1 catalyzed O-dealkylation 2-methoxyphenol produce catechol. other ortho-substituted phenols, including 2-ethoxyphenol, 2-methylphenol (o-cresol) 2-chlorophenol. affinity for these compounds lower than they are less effective at inducing state. Para-substituted meta-substituted ether phenols did not induce spin transition.