Characterization Of Three Glycosidases

This paper mainly concerns the purification and reaction characters of three glycosidases from microorganism Absidia sp.R9g?Absidia sp.G3g and Absidia sp.E9r,and ascertains molecular weight,speciality of purified enzyme and kinetics of enzyme reaction.The there glycosidases were pured using DEAE-cellulose column.The pure enzyme purity and molecular weights of the there glycosidases were determined using SDS-polyacrylamide gel electrophoresis to obtain one spot and M.W.of rutin-a-rhamnosidase?ginsenoside-a-rhamnosidase and icariin glycosidase were 66kDa'58kDa?65kDa.The optimum reaction characters of enzymes shows that optimum condition:rutin-a-rhamnosidase:pH5.0,the range of stabilization is pH 3.0?7.0;temperature,50?,the range of stabilization is 30?60 ?;The rutin-?-rhamnosidase isn't effected by Na+?K+?Ca2+?Mg2+?Zn2+?inhibited by Fe3+?Cu2+;The Vmax is 6.849mmol/L-h,the Km is 123.904mmol/L.ginsenoside-a-rhamnosidase:pH5.0,the range of stabilization is pH 3.0?6.0;temperature,50 ?,the range of stabilization is 30?60 ?;The ginsenoside-a-rhamnosidase isn't effected by Na+?K+?Mg2+?Ca2+,inhibited by Zn2+?Fe3+?Cue+;The Vmax is 0.691mmol/L-h,the Km is 8.918mmol/L.icariin glycosidase:pH4.0,the range of stabilization is pH 3.0-6.0;temperature,50 ?,the range of stabilization is 30?60 ?;The icariin glycosidase isn't effected by Na+?K+?Ca2+?Mg2+?Zn2+?inhibited by Fe3+?Cu2+;The Vmax is 3.012mmol/L·h,the Km is 58.59mmol/L.The enzyme reaction characteristics of the three glycosidases show that rutin-?-rhamnosidase is capable of hydrolyzing the ?-rhamnoside of rutin?hesperidin and naringin?ginsenoside-a-rhamnosidase is capable of hydrolyzing the 6-C,a-rhamnoside of ginsenoside Re and icariin glycosidase is capable of hydrolyzing the glucoside.