Crystallization and preliminary X-ray studies of azoreductases from Bacillus sp. B29.
Crystallization and preliminary X-ray studies of azoreductases from Bacillus sp. B29.
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DOI：
10.1107/s1744309110007785
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发表时间：
2010-05
期刊：
影响因子：
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通讯作者：
D. Ogata;T. Ooi;T. Fujiwara;S. Taguchi;I. Tanaka;M. Yao
中科院分区：
文献类型：
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作者：
D. Ogata;T. Ooi;T. Fujiwara;S. Taguchi;I. Tanaka;M. Yao
来自芽孢杆菌B29的偶氮还原酶是nadh依赖的黄酶，它含有一个黄素单核苷酸（FMN）作为假基，以23 kDa亚基组成的同型二聚体存在。这些酶通过乒乓机制催化各种偶氮化合物的还原性降解。为了确定偶氮染料还原机理的结构-功能关系，对偶氮还原酶进行了x射线晶体学研究。硒代蛋氨酸标记AzrA （SeMet-AzrA）和AzrC采用悬垂气相扩散法结晶。该SeMet-AzrA晶体经2.0 A衍射后归属于P2(1)2(1)2(1)空间群，晶胞参数A = 56.9, b = 69.0, c = 105.4 A。AzrC的原生晶体属于C2空间群，晶胞参数a = 192.0, b = 56.6, c = 105.5 a, β = 115.7°，衍射分辨率为2.21 a。
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 A resolution and was determined to belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 56.9, b = 69.0, c = 105.4 A. The native crystals of AzrC belonged to space group C2, with unit-cell parameters a = 192.0, b = 56.6, c = 105.5 A, beta = 115.7 degrees , and diffracted to 2.21 A resolution.