Open AccessExpression, purification, crystallization and preliminary X‐ray analysis of para ‐nitrophenol 4‐monooxygenase from Pseudomonas putida DLL‐E4
Author(s) -
Liu Weidong,
Shen Wenjing,
Zhao Xiaoli,
Cao Hui,
Cui Zhongli
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109032370
Subject(s) - nitrophenol , crystallization , monooxygenase , pseudomonas putida , chemistry , stereochemistry , escherichia coli , enzyme , biochemistry , organic chemistry , cytochrome p450 , gene , catalysis
Para ‐nitrophenol 4‐monooxygenase (PnpA) plays an important role in bacterial degradation of para ‐nitrophenol by oxidative release of the nitro group from the aromatic ring to form p ‐benzoquinone. In order to understand the structural basis of the function of this enzyme, PnpA was cloned, expressed in Escherichia coli and purified. PnpA was crystallized by the hanging‐drop vapour‐diffusion technique with PEG 4000 as precipitant. The PnpA crystals belonged to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 54.47, b = 77.56, c = 209.17 Å, and diffracted to 2.24 Å resolution.