与L-山梨糖或NADPH复合的费氏葡萄糖杆菌L-山梨糖还原酶的纯化、结晶及初步X射线分析。
Purification, crystallization and preliminary X-ray analysis of L-sorbose reductase from Gluconobacter frateurii complexed with L-sorbose or NADPH.
作者信息
Kubota Keiko, Nagata Koji, Miyazono Ken-ichi, Toyama Hirohide, Matsushita Kazunobu, Tanokura Masaru
机构信息
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, 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. 2009 Jun 1;65(Pt 6):562-4. doi: 10.1107/S1744309109014687. Epub 2009 May 22.
NADPH-dependent L-sorbose reductase (SR) from Gluconobacter frateurii was expressed in Escherichia coli, purified and crystallized with L-sorbose or NADPH using the sitting-drop vapour-diffusion method at 293 K. Crystals of the SR-L-sorbose complex and the SR-NADPH complex were obtained using reservoir solutions containing PEG 2000 or PEG 400 as precipitants and diffracted X-rays to 2.38 and 1.90 A resolution, respectively. The crystal of the SR-L-sorbose complex belonged to space group C222(1), with unit-cell parameters a = 124.2, b = 124.1, c = 60.8 A. The crystal of the SR-NADPH complex belonged to space group P2(1), with unit-cell parameters a = 124.3, b = 61.0, c = 124.5 A, beta = 89.99 degrees . The crystals contained two and eight molecules, respectively, in the asymmetric unit.
从弗氏葡萄糖杆菌中提取的依赖烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的L-山梨糖还原酶（SR）在大肠杆菌中表达，经纯化后，于293K使用坐滴气相扩散法，与L-山梨糖或NADPH一起进行结晶。使用含有聚乙二醇2000（PEG 2000）或聚乙二醇400（PEG 400）作为沉淀剂的储液，分别获得了SR-L-山梨糖复合物和SR-NADPH复合物的晶体，其X射线衍射分辨率分别为2.38 Å和1.90 Å。SR-L-山梨糖复合物的晶体属于空间群C222(1)，晶胞参数为a = 124.2 Å，b = 124.1 Å，c = 60.8 Å。SR-NADPH复合物的晶体属于空间群P2(1)，晶胞参数为a = 124.3 Å，b = 61.0 Å，c = 124.5 Å，β = 89.99°。非对称单元中分别包含两个和八个分子。
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