Open AccessResolution of the 4‐hydroxy‐3‐methylbenzoate hydroxylase of Pseudomonas putida into two protein components
Author(s) -
ElMansi E.M.T.,
Hopper D.J.
Publication year - 1990
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1990.tb03987.x
Subject(s) - hydroxylation , chemistry , oxygenase , pseudomonas putida , reductase , stereochemistry , enzyme , biochemistry
4‐Hydroxy‐3‐methylbenzoate hydroxylase of Pseudomonas putida was found to be inducible, rather than constitutive, extremely unstable and requires an electron donor (NADH or NADPH) and molecular oxygen for activity, suggesting a mono‐oxygenase type of enzyme. It was resolved into two protein components by ion‐exchange chromatography. The first protein component was identified as an NADH: acceptor (DCIP) oxido‐reductase, with the second catalysing the hydroxylation reaction. A mechanism illustrating the role of each reaction of this mono‐oxygenase in the hydroxylation reaction is proposed.