Crystallization and preliminary X-ray diffraction analysis of tetrathionate hydrolase from Acidithiobacillus ferrooxidans.
Crystallization and preliminary X-ray diffraction analysis of tetrathionate hydrolase from Acidithiobacillus ferrooxidans.
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DOI：
10.1107/s1744309113013419
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发表时间：
2013-06
期刊：
影响因子：
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通讯作者：
T. Kanao;M. Kosaka;K. Yoshida;Hisayuki Nakayama;T. Tamada;R. Kuroki;H. Yamada;J. Takada;K. Kamim
中科院分区：
文献类型：
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作者：
T. Kanao;M. Kosaka;K. Yoshida;Hisayuki Nakayama;T. Tamada;R. Kuroki;H. Yamada;J. Takada;K. Kamim
来自铁和硫氧化细菌Acidithiobacillusferrooxidans的连四硫酸盐水解酶（4THase）催化连四硫酸盐不成比例水解为元素硫、硫代硫酸盐和硫酸盐。将编码4TH酶的基因（Af-tth）在大肠杆菌中以包涵体形式表达。重组Af-Tth通过在酸性条件下重折叠而活化，然后纯化至均一。使用悬滴气相扩散法在含有50 mM氯化钠和33%（v/v）PEG 1000的20 mM甘氨酸缓冲液（pH 10）中结晶重组蛋白。晶体为六方柱，尺寸为0.2 × 0.05 × 0.05 mm，X射线衍射分辨率为2.15 nm，属P3（1）或P3（2）空间群，晶胞参数a = B = 92.1，c = 232.6 nm。
Tetrathionate hydrolase (4THase) from the iron- and sulfur-oxidizing bacterium Acidithiobacillus ferrooxidans catalyses the disproportionate hydrolysis of tetrathionate to elemental sulfur, thiosulfate and sulfate. The gene encoding 4THase (Af-tth) was expressed as inclusion bodies in recombinant Escherichia coli. Recombinant Af-Tth was activated by refolding under acidic conditions and was then purified to homogeneity. The recombinant protein was crystallized in 20 mM glycine buffer pH 10 containing 50 mM sodium chloride and 33%(v/v) PEG 1000 using the hanging-drop vapour-diffusion method. The crystal was a hexagonal cylinder with dimensions of 0.2 × 0.05 × 0.05 mm. X-ray crystallographic analysis showed that the crystal diffracted to 2.15 Å resolution and belongs to space group P3(1) or P3(2), with unit-cell parameters a = b = 92.1, c = 232.6 Å.