Chu Huy Nhat, Kobayashi Jun, Yoshikane Yu, Mikami Bunzo, Yagi Toshiharu
Faculty of Agriculture and Agricultural Science Program, Graduate School of Integral Arts and Science, Kochi University, Monobe, Nankoku, Kochi 783-8502, Japan.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Jun 1;66(Pt 6):718-20. doi: 10.1107/S1744309110015101. Epub 2010 May 29.
Pyridoxal 4-dehydrogenase from Mesorhizobium loti MAFF303099 was overexpressed in Escherichia coli. The recombinant selenomethionine-substituted enzyme was purified and crystallized by the sitting-drop vapour-diffusion method using PEG 4000 as precipitant. Crystals grew in the presence of 0.45 mM NAD(+). The crystals diffracted to 2.9 A resolution and belonged to the monoclinic space group P2(1), with unit-cell parameters a = 86.20, b = 51.11, c = 91.73 A, beta = 89.36 degrees. The calculated V(M) values suggested that the asymmetric unit contained four molecules.
来自百脉根中生根瘤菌MAFF303099的吡哆醛4-脱氢酶在大肠杆菌中过表达。使用聚乙二醇4000作为沉淀剂，通过坐滴气相扩散法对重组硒代蛋氨酸取代酶进行纯化和结晶。晶体在0.45 mM NAD(+)存在下生长。晶体衍射分辨率达到2.9 Å，属于单斜空间群P2(1)，晶胞参数为a = 86.20、b = 51.11、c = 91.73 Å，β = 89.36°。计算得到的V(M)值表明不对称单元包含四个分子。
