원문정보
초록
영어
The trehalose phosphorylase (TPase, EC 2.4.1.64) and kojibiose phosphorylase (KPase, EC 2.4.1.230) catalyze the phospholytic activities on the α1,1glucosidic linkage and α 1,2glucosidic linkage in each substrates (trehalose and kojibiose), respectively. Both enzymes also perform the reverse reaction of the phosphorylase, with which α1,1 and α 1,2- glucosidic linkages are formed with glucose and glucose-6-phosphate as substrates. In this study, the KPase and TPase genes were expressed in E. coli. Two phosphorylases, TPase and KPase from each Anaerocellum thermophilum DSM 6725 and Pyrococcus sp. ST04 were incorporated into E. coli BL21, respectively. The presence of catalytic activities of both recombinant enzymes were confirmed. The continuous reaction with TPase and KPase was carried out with trehalose as a sole substrate. The thin layer chromatography (TLC) and high performance anion exchange chromatography (HPAEC) analysis revealed the formation of an unusual trisaccharide in a reaction mixture. The structural analysis of an unusual trisaccharide showed that the final product was a rare tripolysaccharide, selaginose. The result indicated that a rare tripolysaccharide, selaginose, can be synthesized by one-step enzymatic reaction system with KPase and TPase.