短双歧杆菌木酮糖-5-磷酸/果糖-6-磷酸磷酸酮醇酶的过表达、结晶及初步X射线分析
Overexpression, crystallization and preliminary X-ray analysis of xylulose-5-phosphate/fructose-6-phosphate phosphoketolase from Bifidobacterium breve.
作者信息
Suzuki Ryuichiro, Kim Byung-Jun, Shibata Tsuyoshi, Iwamoto Yuki, Katayama Takane, Ashida Hisashi, Wakagi Takayoshi, Shoun Hirofumi, Fushinobu Shinya, Yamamoto Kenji
机构信息
Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Aug 1;66(Pt 8):941-3. doi: 10.1107/S1744309110023845. Epub 2010 Jul 29.
Abstract
The xylulose-5-phosphate/fructose-6-phosphate phosphoketolase gene from Bifidobacterium breve was cloned and overexpressed in Escherichia coli. The enzyme was purified to homogeneity and crystallized by the sitting-drop vapour-diffusion method. Crystals were obtained at 293 K using 0.05 mM thiamine diphosphate, 0.25 mM MgCl2, 24%(w/v) PEG 6000 and 0.1 M Bicine pH 9.0. The crystals belonged to the tetragonal space group I422, with unit-cell parameters a=b=174.8, c=163.8 A, and diffracted to beyond 1.7 A resolution.
摘要
短双歧杆菌的木酮糖-5-磷酸/果糖-6-磷酸磷酸酮醇酶基因被克隆并在大肠杆菌中过表达。该酶被纯化至同质，并通过坐滴气相扩散法结晶。在293 K下，使用0.05 mM硫胺二磷酸、0.25 mM氯化镁、24%（w/v）聚乙二醇6000和0.1 M二（2-羟乙基）氨基甘氨酸pH 9.0获得晶体。晶体属于四方晶系空间群I422，晶胞参数a=b=174.8，c=163.8 Å，并衍射至超过1.7 Å分辨率。
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