Extraction And Separation Of Oligosaccharides And Ophipogon Japonicus Saponin By Enzyme

Conditions for enzymatic extraction of saponins and oligosaccharides from Ophiopogon were studied.Then Ophiopogon japonicus saponin and Ophiopogon oligosaccharides were isolated and purified.Finally,the composition、molecular weight and nature of Ophiopogon oligosaccharides were Preliminarily analysisrd.The optimal conditions for the enzyme cultivation were explored in:beginning with Absidia sp.08s,inducing by 20%Ophiopogon japonicus in the medium,and cultivating for 6 days at 30℃.Conditions on enzymatic preparation of Ophiopogon oligosaccharides were studied.Through the experiment,the optimal conditions were explored in:beginning with Ophiopogon powder soaked by 4 times volume of buffer,crude enzyme were added,Optimum temperature is 50℃,Optinum pH is 5.reacted 12 hours.80%alcohol were used to separate Oligosaccharides and Ophipogon japonicus Saponin from each other for 12h.Ophiopogon oligosaccharides were in the precipitate,dissolved in water and filter again,crud Ophiopogon oligosaccharides were obtained.Ophipogon japonicus Saponin were in alcohol.The alcohol evaporated dry as crud Ophipogon japonicus Saponin.After purified by AB-8 and D-296 Macroporous Adsorptive Resins,carbohydrate and Pigment were removed,Ophiopogon japonicus Total Saponins were obtained,The yield rate was 0.28%.Several methods were campared to remove protein from Ophiopogon oligosaccharides samle.Sevage-TCA method was found to be the best,with the protein removal rate of 81.18%and a polysaccharide loss rate of 8.39%.Isolated by Sephadex G-100,OP-Ⅰ、OP-Ⅱ、OP-Ⅲ were obtained from Ophipogon Oligosaccharides estimating by gel filtration,their relative molecular weights were approximately determined as 39kDa、7kDa、2kDa,respectively.Analyzed by HPLC,the monosaccharide compositions of OP-Ⅰ、OP-Ⅱ were fructose,while the monosaccharide compositions of OP-III were glucose and fructose.