Open AccessThe purification, crystallization and preliminary structural characterization of FAD‐dependent monooxygenase PhzS, a phenazine‐modifying enzyme from Pseudomonas aeruginosa
Author(s) -
Gohain Neelakshi,
Thomashow Linda S.,
Mavrodi Dmitri V.,
Blankenfeldt Wulf
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106034464
Subject(s) - phenazine , pyocyanin , derepression , monooxygenase , chemistry , pseudomonas aeruginosa , stereochemistry , enzyme , biochemistry , bacteria , biology , virulence , quorum sensing , psychological repression , gene expression , genetics , cytochrome p450 , gene
The blue chloroform‐soluble bacterial metabolite pyocyanin (1‐hydroxy‐5‐methyl‐phenazine) contributes to the survival and virulence of Pseudomonas aeruginosa , an important Gram‐negative opportunistic pathogen of humans and animals. Little is known about the two enzymes, designated PhzM and PhzS, that function in the synthesis of pyocyanin from phenazine‐1‐carboxylic acid. In this study, the FAD‐dependent monooxygenase PhzS was purified and crystallized from lithium sulfate/ammonium sulfate/sodium citrate pH 5.5. Native crystals belong to space group C 2, with unit‐cell parameters a = 144.2, b = 96.2, c = 71.7 Å, α = γ = 90, β = 110.5°. They contain two monomers of PhzS in the asymmetric unit and diffract to a resolution of 2.4 Å. Seleno‐ l ‐methionine‐labelled PhzS also crystallizes in space group C 2, but the unit‐cell parameters change to a = 70.6, b = 76.2, c = 80.2 Å, α = γ = 90, β = 110.5° and the diffraction limit is 2.7 Å.