Vorgias C E, Kingswell A J, Dauter Z, Oppenheim A B
European Molecular Biology Laboratory, DESY, Hamburg, Germany.
J Mol Biol. 1992 Aug 5;226(3):897-8. doi: 10.1016/0022-2836(92)90640-6.
The chiA gene encoding for the chitinase enzyme from Serratia marcescens was efficiently overexpressed under the pL promoter and the enzyme was secreted into the growth medium. The chitinase was purified to homogeneity using affinity chromatography on a Phenyl-Sepharose column and the protein was successfully crystallized. The crystals are presently in the form of small needles in space group C222(1) and have unit cell dimensions a = 204(+/- 0.5) A, b = 134(+/- 0.5) A, c = 60(+/- 0.5) A. The crystals diffract X-rays to about 3 A resolution and are suitable for three-dimensional structural analysis.
编码粘质沙雷氏菌几丁质酶的chiA基因在pL启动子的作用下高效过表达，且该酶分泌到生长培养基中。使用苯基琼脂糖柱上的亲和色谱法将几丁质酶纯化至同质，该蛋白质成功结晶。目前晶体呈C222(1)空间群中的小针状，晶胞参数为a = 204(±0.5) Å，b = 134(±0.5) Å，c = 60(±0.5) Å。这些晶体的X射线衍射分辨率约为3 Å，适用于三维结构分析。
