Dos Reis Caio Vinicius, Bernardes Amanda, Polikarpov Igor
Instituto de Física de São Carlos, Universidade de São Paulo, Trabalhador São Carlense 400, 13566-590 São Carlos-SP, Brazil.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 May 1;69(Pt 5):588-91. doi: 10.1107/S174430911301110X. Epub 2013 Apr 30.
Xylose isomerase (EC 5.3.1.5) is a key enzyme in xylose metabolism which is industrially important for the transformation of glucose and xylose into fructose and xylulose, respectively. The Bifidobacterium adolescentis xylA gene (NC_008618.1) encoding xylose isomerase (XI) was cloned and the enzyme was overexpressed in Escherichia coli. Purified recombinant XI was crystallized using the sitting-drop vapour-diffusion method with polyethylene glycol 3350 as the precipitating agent. A complete native data set was collected to 1.7 Å resolution using a synchrotron-radiation source. The crystals belonged to the orthorhombic space group P21212, with unit-cell parameters a = 88.78, b = 123.98, c = 78.63 Å.
木糖异构酶(EC 5.3.1.5)是木糖代谢中的关键酶,在工业上分别将葡萄糖和木糖转化为果糖和木酮糖具有重要意义。克隆了编码木糖异构酶(XI)的青春双歧杆菌木糖A基因(NC_008618.1),并在大肠杆菌中对该酶进行了过量表达。使用聚乙二醇3350作为沉淀剂,通过坐滴气相扩散法对纯化的重组XI进行结晶。使用同步辐射源收集了完整的天然数据集,分辨率达到1.7 Å。晶体属于正交空间群P21212,晶胞参数a = 88.78,b = 123.98,c = 78.63 Å。