A Soluble Cytochrome P-450 Functional in Methylene Hydroxylation

Abstract

1. A methylene hydroxylase system from camphor induced Pseudomonas putida strain C1 cells has been separated into three fractions: a putidaredoxin reductase, putidaredoxin (an iron-sulfur protein), and a hydroxylase (shown to be a soluble cytochrome P-450). 2. This system of enzymes catalyzes the hydroxylation of methylene carbon 5 of camphor with reduced disphosphopyridine nucleotide as a primary electron donor and molecular oxygen as acceptor. 3. The redoxin serves as the electron carrier from the reduced pyridine nucleotide and reductase to the cytochrome P-450-substrate complex. 4. The coupling of the hydroxylase (P-450) to the reduced diphosphopyridine nucleotide dehydrogenase (reductase), and the putidaredoxin is demonstrated, and substrate is shown to be required for the over-all reaction.