Putidaredoxin Reductase Research Articles

Current status of the sequence studies of the Pseudomonas putida camphor hydroxylase system.

A methylene hydroxylase system from camphor induced Pseudomonas putida has been separated into a putidaredoxin reductase, putidaredoxin (an iron-sulfur protein), and a hydroxylase fraction known as soluble cytochrome P-450 (1). This mixed function oxidase catalyzes the hydroxylation of methylene carbon 5 of camphor with reduced diphosphopyridine nucleotide as the primary electron donor and with molecular oxygen as the acceptor. The physicochemical and kinetic properties of the hydroxylase system are discussed in review articles by Gunsalus et al (2,3 and see articles by Gunsalus and Sligar in this book).

A Soluble Cytochrome P-450 Functional in Methylene Hydroxylation

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.