Separation And Purification Of Gypenoside From Gynostemma Pentaphyllum

This thesis mainly from the Chinese herbal medicine extraction of gynostemma pentaphyllum gynostemma pentaphyllum saponins,and the column chromatography total saponins of gynostemma pentaphyllum was isolated and purified.And then to microbial enzymes gynostemma pentaphyllum saponins transformation for the purpose,and screened gynostemma pentaphyllum saponins indican the generation of enzyme strains,and optimize the fermentation conditions.The last of gynostemma pentaphyllum saponins glycosidase enzymes properties were studied.The single factor experiment method optimization gynostemma pentaphyllum saponins of water extraction process,determine the boiled law to extract gynostemma pentaphyllum thick saponins is the best technological conditions of material liquid proportion of 20:1,three hours and boiled water three times,in the condition of the total saponins gynostemma pentaphyllum extraction rate is 5.4%.With AB-8 macroporous adsorption resin to take off the sugar,D-296 ion exchange resin column decoloring and silica gel column chromatography of gynostemma pentaphyllum thick saponins separation and purification.Gynostemma pentaphyllum thick saponins after AB-8 macroporous adsorption resin to take off the sugar,D-296 ion exchange resin column after bleaching,the total saponins of gynostemma pentaphyllum,the rate is 52.4%.Silica gel column chromatography separation gynostemma pentaphyllum total saponins,in proportion with V(chloroform):V(methanol)= 8.5:1.5 comes saponins sample A and B,rates were 1.2%,2.5%;In proportion with V(chloroform):V(methanol)= "were saponins samples C,yield is 16.3%.With the polysaccharide gynostemma pentaphyllum saponins samples C as the substrate,from 7 kinds of bacteria can produce were gynostemma pentaphyllum saponins indican strains enzymes,and finally determined that Absidia sp.GYP4r strains for enzyme production strains,the bacteria in 30 ? fermentation cultivation under the condition of six days have been enzyme hydrolysis of liquid to substrate ability is the best;The best thing for 20%gynostemma pentaphyllum induction.Gynostemma pentaphyllum saponins glycosidase enzymes that the optimal pH and temperature are pH5 and 50 ? respectively,this enzyme in pH4?6 range in 60 ? below and stable.Na+?K+?Mg2+?Zn2+?Ca2+ five ion to little effect on the enzyme,Fe3+?Cu2+ two ion to effect of the enzyme of vigor has obvious inhibition;The enzyme reaction dynamics research shows that Vmax is 3.3 mmol/(L·h)and Km is 1.36 mmol/L.The relative molecular mass 75.9 kDa.