title: A phthalate family oxygenase reductase supports terpene alcohol oxidation by CYP238A1 from Pseudomonas putida KT2440.
abstract: CYP238A1, one of the two P450 enzymes in the genome of Pseudomonas putida KT2440, has been produced heterologously in Escherichia coli, purified, and found to bind acyclic and cyclic terpene alcohols such as farnesol, nerolidol, linalool, and terpineol. The other P450 enzyme in this organism (gene locus: PP1950) was also produced in E. coli but no substrate has been identified from a limited screen. A phthalate family oxygenase reductase (PFOR) encoded by the PP1957 gene, just downstream of the PP1955 gene for CYP238A1, accepts electrons from the reduced form of both nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate and is able to support monooxygenase activity of CYP238A1, both in vitro and in E. coli, in which both enzymes are produced. CYP238A1 oxidizes cis- and trans-nerolidol to the 9-hydroxy product, with no evidence of attack at the olefinic double bonds. The NADH turnover rate of 170 nmol(nmol-P450)⁻¹ Min⁻¹ for CYP238A1 with cis-nerolidol as substrate at a PP1957:CYP238A1 concentration ratio of 8:1 suggests that this PFOR could function as the physiological redox partner for CYP238A1. The physiological role of CYP238A1 may be related to the PP1955 gene being part of an island/cluster of inducible genes associated with energy metabolism and response to xenobiotics.
authors.affiliation: School of Chemistry and Physics, The University of Adelaide, Adelaide, SA, Australia. stephen.bell@adelaide.edu.au
authors.forename: Stephen Graham
authors.initials: SG
authors.lastname: Bell
authors.forename: Laura
authors.initials: L
authors.lastname: French
authors.forename: Nicholas Huw
authors.initials: NH
authors.lastname: Rees
authors.forename: Sophia Shuyi
authors.initials: SS
authors.lastname: Cheng
authors.forename: Gail
authors.initials: G
authors.lastname: Preston
authors.forename: Luet-Lok
authors.initials: LL
authors.lastname: Wong
mesh_terms: D000438:Alcohols / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D016958:Pseudomonas putida / Q000201:enzymology*; D013729:Terpenes / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000438:Alcohols; D013729:Terpenes; D010088:Oxidoreductases; C049135:phthalate oxygenase reductase