来源于大肠杆菌的6-磷酸己酮糖合成酶和6-磷酸果糖异构酶的原核表达与活性测定

Prokaryotic Expression and Activity Measurement of 3-hexulose-6-p hosphate Synthase and 6-phospho-3-hexuloisomerase from Escherichia coli

同源性搜索显示大肠杆菌中存在核酮糖单磷酸途径关键酶6-磷酸己酮糖合成酶(HPS)和6-磷酸果糖异构酶(PHI)基因的同源序列,但没有其相关活性和功能方面的报道。本研究利用PCR方法从大肠杆菌的基因组中扩增hps和phi的同源序列(分别简称为Ehps和Ephi),构建了大肠杆菌Ehps和Ephi基因的原核表达载体pDEST17-Ehps和pDEST17-Ephi,表达和纯化了重组蛋白EHPS和EPHI,酶活性检测结果表明EHPS和EPHI具有活性。

Homology search shows that there are homologous sequences of 3-hexulose-6-phosphate synthase （HPS） and 6-phos- pho-3-hexuloisomerase （PHI） , both of which involve in Ribulose monophosphate pathway in Escherichia coli, but there is no report on the activities and functions of the enzymes. The hps and phi homologous sequences designated as Ehps and Ephi, were amplified from E. coli genome by PCR to construct the prokaryotic expression vectors pDEST17-Ehps and pDEST17-Ephi, respectively. The recombi- nant proteins encoded by the two genes were expressed and partially purified, Further analysis indicated that the recombinant protein exhibited enzyme activities.