巨大芽孢杆菌WSH-002丙氨酸脱氢酶的结晶及初步X射线分析
Crystallization and preliminary X-ray analysis of an alanine dehydrogenase from Bacillus megaterium WSH-002.
作者信息
Lu Xiaoyun, Yi Qiufen, Zhang Guofang, Zhu Xianming, Zhou Honggang, Dong Hui
机构信息
Tianjin University of Science and Technology, Tianjin 300222, People's Republic of China.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Aug;69(Pt 8):934-6. doi: 10.1107/S1744309113019672. Epub 2013 Jul 27.
Alanine dehydrogenase (L-AlaDH) from Bacillus megaterium WSH-002 catalyses the NAD⁺-dependent interconversion of L-alanine and pyruvate. The enzyme was expressed in Escherichia coli BL21 (DE3) cells and purified with a His6 tag by Ni²⁺-chelating affinity chromatography for X-ray crystallographic analysis. Crystals were grown in a solution consisting of 0.1 M HEPES pH 8.0, 12%(w/v) polyethylene glycol 8000, 8%(v/v) ethylene glycol at a concentration of 15 mg ml⁻¹ purified protein. The crystal diffracted to 2.35 Å resolution and belonged to the trigonal space group R32, with unit-cell parameters a = b = 125.918, c = 144.698 Å.
巨大芽孢杆菌WSH-002的丙氨酸脱氢酶（L-AlaDH）催化L-丙氨酸和丙酮酸之间依赖NAD⁺的相互转化。该酶在大肠杆菌BL21（DE3）细胞中表达，并通过Ni²⁺螯合亲和色谱法用His6标签进行纯化，用于X射线晶体学分析。晶体在由0.1 M HEPES pH 8.0、12%（w/v）聚乙二醇8000、8%（v/v）乙二醇组成的溶液中生长，纯化蛋白浓度为15 mg ml⁻¹。晶体衍射分辨率达到2.35 Å，属于三方晶系空间群R32，晶胞参数a = b = 125.918，c = 144.698 Å。
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