Sucrose phosphorylase(SPase)catalyzes the conversion of sucrose to α-Glucose 1-phosphate(G-1-P)and fructose.G-1-P functions as a glycosyl donor and is an initial substrate for biosynthesis cascades.This study charact...Sucrose phosphorylase(SPase)catalyzes the conversion of sucrose to α-Glucose 1-phosphate(G-1-P)and fructose.G-1-P functions as a glycosyl donor and is an initial substrate for biosynthesis cascades.This study characterized the enzymatic properties of a novel SPase from Bifidobacterium dentium(BdSP)and explored its potential as an initial enzyme in cascade reactions.BdSP displayed 91-93%homology with the SPase from Bifidobacterium species.Recombinant BdSP was characterized as a dimer with optimal temperatures of 55℃and 60℃and pH of 7.0 and 5.0 for the phosphorolysis and synthesis reactions,respectively.The melting temperature of BdSP was 68.91℃,and its half-life was 235.1 min at 65℃.Kinetic studies demonstrated that BdSP can produce G-1-P from sucrose more efficiently than other SPases,with a production rate of 68%at 55℃ for 2 h.Furthermore,the cascade reaction of BdSP and cellobiose phosphorylase from Clostridium thermocellum YM4(CtCBP)reached an effective production of cellobiose(80.4%)at 55℃for 18 h.展开更多
基金supported by a National Research Foundation of Korea(NRF)grant funded by the Korean government(MSIT,Ministry of Science and ICT)(grant no.2021R1A4A1023437)the Ministry of Education(RS-2023-00239622).
文摘Sucrose phosphorylase(SPase)catalyzes the conversion of sucrose to α-Glucose 1-phosphate(G-1-P)and fructose.G-1-P functions as a glycosyl donor and is an initial substrate for biosynthesis cascades.This study characterized the enzymatic properties of a novel SPase from Bifidobacterium dentium(BdSP)and explored its potential as an initial enzyme in cascade reactions.BdSP displayed 91-93%homology with the SPase from Bifidobacterium species.Recombinant BdSP was characterized as a dimer with optimal temperatures of 55℃and 60℃and pH of 7.0 and 5.0 for the phosphorolysis and synthesis reactions,respectively.The melting temperature of BdSP was 68.91℃,and its half-life was 235.1 min at 65℃.Kinetic studies demonstrated that BdSP can produce G-1-P from sucrose more efficiently than other SPases,with a production rate of 68%at 55℃ for 2 h.Furthermore,the cascade reaction of BdSP and cellobiose phosphorylase from Clostridium thermocellum YM4(CtCBP)reached an effective production of cellobiose(80.4%)at 55℃for 18 h.
