Mugo Andrew Njagi, Kobayashi Jun, Mikami Bunzo, Ohnishi Kouhei, Yagi Toshiharu
Faculty of Agriculture and Agricultural Science Program, Graduate School of Integral Arts and Science, Kochi University, Nankoku, Kochi 783-8502, Japan.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Jan 1;68(Pt 1):66-8. doi: 10.1107/S1744309111048688. Epub 2011 Dec 24.
Vitamin B(6)-degradation pathway I has recently been identified in Mesorhizobium loti MAFF303099. Pyridoxine 4-oxidase, an FAD-dependent enzyme, is the first enzyme in this pathway and catalyzes the irreversible oxidation of pyridoxine to pyridoxal. The enzyme was overexpressed in Escherichia coli with a His(6) tag and purified. The recombinant enzyme was crystallized at 277 K by the sitting-drop vapour-diffusion method using PEG 4000 as the precipitant. The crystal, which belonged to space group P2(1)2(1)2(1) with unit-cell parameters a = 62.38, b = 79.44, c = 136.43 Å, diffracted to 2.2 Å resolution. The calculated V(M) value (3.19 Å(3) Da(-1)) suggested that the asymmetric unit contained one molecule.
维生素B6降解途径I最近在百脉根中生根瘤菌MAFF303099中被发现。吡哆醇4-氧化酶是一种依赖黄素腺嘌呤二核苷酸的酶，是该途径中的第一种酶，催化吡哆醇不可逆地氧化为吡哆醛。该酶在带有His(6)标签的大肠杆菌中过表达并纯化。使用聚乙二醇4000作为沉淀剂，通过坐滴气相扩散法在277 K下使重组酶结晶。该晶体属于空间群P2(1)2(1)2(1)，晶胞参数a = 62.38、b = 79.44、c = 136.43 Å，衍射分辨率达到2.2 Å。计算得到的V(M)值（3.19 Å(3) Da(-1)）表明不对称单元包含一个分子。
