Open AccessCrystallization and preliminary X‐ray studies of azoreductases from Bacillus sp. B29
Author(s) -
Ogata Daiki,
Ooi Toshihiko,
Fujiwara Takaaki,
Taguchi Seiichi,
Tanaka Isao,
Yao Min
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110007785
Subject(s) - flavin mononucleotide , crystallization , chemistry , crystallography , resolution (logic) , stereochemistry , enzyme , flavin group , biochemistry , organic chemistry , artificial intelligence , computer science
Azoreductases from Bacillus sp. B29 are NADH‐dependent flavoenzymes which contain a flavin mononucleotide (FMN) as a prosthetic group and exist as homodimers composed of 23 kDa subunits. These enzymes catalyze the reductive degradation of various azo compounds by a ping‐pong mechanism. In order to determine the structure–function relationship of the azo‐dye reduction mechanism, an X‐ray crystallographic study of azoreductases was performed. Selenomethionine‐labelled AzrA (SeMet‐AzrA) and AzrC were crystallized by the hanging‐drop vapour‐diffusion method. A crystal of SeMet‐AzrA diffracted to 2.0 Å resolution and was determined to belong to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 56.9, b = 69.0, c = 105.4 Å. The native crystals of AzrC belonged to space group C 2, with unit‐cell parameters a = 192.0, b = 56.6, c = 105.5 Å, β = 115.7°, and diffracted to 2.21 Å resolution.