Open AccessCrystallization and preliminary X‐ray analysis of an alanine dehydrogenase from Bacillus megaterium WSH‐002
Author(s) -
Lu Xiaoyun,
Yi Qiufen,
Zhang Guofang,
Zhu Xianming,
Zhou Honggang,
Dong Hui
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113019672
Subject(s) - bacillus megaterium , polyethylene glycol , crystallization , ethylene glycol , escherichia coli , dehydrogenase , alanine , nad+ kinase , chemistry , enzyme , nuclear chemistry , biochemistry , biology , amino acid , bacteria , organic chemistry , gene , genetics
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 His 6 tag by Ni 2+ ‐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 R 32, with unit‐cell parameters a = b = 125.918, c = 144.698 Å.