来自土壤杆菌属KY5R菌株的尼龙寡聚体水解酶（NylC）的结晶及X射线衍射分析
Crystallization and X-ray diffraction analysis of nylon-oligomer hydrolase (NylC) from Agromyces sp. KY5R.
作者信息
Yasuhira Kengo, Shibata Naoki, Tanaka Yasuhito, Kumagai Naoya, Tanaka Yusuke, Nagai Keisuke, Kato Dai-ichiro, Takeo Masahiro, Negoro Seiji, Higuchi Yoshiki
机构信息
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo, Himeji, Hyogo, Japan.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Aug 1;67(Pt 8):892-5. doi: 10.1107/S1744309111022858. Epub 2011 Jul 19.
6-Aminohexanoate-oligomer hydrolase (NylC) from Agromyces sp. KY5R was expressed in Escherichia coli JM109 and purified by ammonium sulfate fractionation, anion-exchange column chromatography and gel-filtration chromatography. NylC was crystallized by the sitting-drop vapour-diffusion method with sodium citrate as a precipitant in 0.1 M HEPES buffer pH 7.5 containing 0.2 M NaCl. Diffraction data were collected from native and K(2)PtCl(4)-derivative crystals to resolutions of 2.00 and 2.20 Å, respectively. The obtained crystal was plate-shaped, with an I-centred orthorhombic space group and unit-cell parameters a = 155.86, b = 214.45, c = 478.80 Å. The anomalous difference Patterson map of the K(2)PtCl(4)-derivative crystal suggested that the space group was I222 rather than I2(1)2(1)2(1).
来自土壤杆菌属KY5R菌株的6-氨基己酸酯寡聚体水解酶（NylC）在大肠杆菌JM109中表达，并通过硫酸铵分级沉淀、阴离子交换柱色谱和凝胶过滤色谱进行纯化。NylC采用坐滴气相扩散法，以柠檬酸钠作为沉淀剂，在含有0.2 M NaCl的0.1 M HEPES缓冲液（pH 7.5）中结晶。分别从天然晶体和K₂PtCl₄衍生物晶体收集衍射数据，分辨率分别为2.00 Å和2.20 Å。获得的晶体呈板状，空间群为体心正交，晶胞参数a = 155.86、b = 214.45、c = 478.80 Å。K₂PtCl₄衍生物晶体的反常差值帕特森图表明，空间群为I222而非I2₁2₁2₁。
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