Xing Keke, Gan Weiqiong, Jia Minze, Gao Feng, Gong Weimin
Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Beijing 100101, People's Republic of China.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Jun;69(Pt 6):666-8. doi: 10.1107/S1744309113012128. Epub 2013 May 25.
The flavoenzyme fructosyl peptide oxidase (FPOX) catalyses the oxidative deglycation of fructosyl amino acids or fructosyl dipeptides to produce amino acids, glucosone and hydrogen peroxide. In this study, FPOX protein from Eupenicillium terrenum sp. (EtFPOX) was expressed in Escherichia coli and purified by Ni-affinity and gel-filtration chromatography. EtFPOX crystals were obtained using the sitting-drop vapour-diffusion method with polyethylene glycol 3350 as precipitant. X-ray diffraction data were collected to 1.90 Å resolution using a synchrotron-radiation source. The crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 65.6, b = 80.0, c = 83.4 Å, and contained one molecule in the asymmetric unit. The calculated Matthews coefficient and solvent content were 2.22 Å(3) Da(-1) and 44.62%, respectively.
黄素酶果糖基肽氧化酶（FPOX）催化果糖基氨基酸或果糖基二肽的氧化脱糖基化反应，生成氨基酸、葡糖酮和过氧化氢。在本研究中，来自土生正青霉（EtFPOX）的FPOX蛋白在大肠杆菌中表达，并通过镍亲和层析和凝胶过滤层析进行纯化。使用聚乙二醇3350作为沉淀剂，通过坐滴气相扩散法获得了EtFPOX晶体。使用同步辐射源收集了分辨率为1.90 Å的X射线衍射数据。晶体属于空间群P2(1)2(1)2(1)，晶胞参数a = 65.6、b = 80.0、c = 83.4 Å，不对称单元中含有一个分子。计算得到的马修斯系数和溶剂含量分别为2.22 Å(3) Da(-1)和44.62%。
