Study On Production Of High Purity Stachyose From Stachys Sieboldii Miq

Stachyose is a natural occurring functional oligosaccharides,with high stability,low sweetness and calorie.It possessed function of significantly promoting the proliferation of bifidobacteria,lactobacillus and other beneficial bacteria,rapidly improving human digestive tract environment,regulating microbial flora balance,known as the "natural super bifidus factor".With the rapid development of the world economy and people pay more and more attention to health,stachyose received wide attention in the food ingredients,it's demand has grown increasingly in the world market.Stachys sieboldii Miq,also known as Chinese artichoke,widely distributed in the China's Jiangsu,Shandong,Hubei,Ningxia,Gansu and Chongqing province.The moisture content of fresh Stachys sieboldii Miq about is about 78%,followed by stachyose,the content of 12%-15%,it is the preferred raw material for the preparation of stachyose.This paper used the Waters XBridge Amide chromatography column(4.6 mm×250 mm,3.5?m)with high performance liquid chromatography and evaporative light-scattering detector(HPLC-ELSD)to optimize the method for the determination of oligosaccharide in Stachys Sieboldii Miq.The optimum testing conditions were as follows:mobile phase was acetonitrile(A)-water(B),0.2%(V/V)triethylamine was added,gradient elution(A% 70%?45%?70%? 70%,the corresponding time period was 0 min?16 min?16.01 min?20 min),the flow rate was 1.0mL/min and the column temperature was 35?.The temperature of evaporative light-scattering detector drift tube was 95?,and carrier gas flow rate was 2.5L/min.The method has high sensitivity,good repeatability and recovery,can accurately and quickly detected the contents of sucrose,raffinose and stachyose in Stachys Sieboldii Miq.Analysed the basic components of Stachys Sieboldii Miq from different place of origin,the Stachys Sieboldii Miq from Gansu province has the highest stachyose content,thus to be the best raw materials of stachyose.Its dry powder contains 59.88% carbohydrates,3.76% ash,0.76% crude fat,15.37% crude protein and 11.02% water.Used the water extract stachyose from Stachys Sieboldii Miq,based on single factor experiment and response surface experiments,in the range of factors and levels,it was proved that solvent-material ratio exhibited the most important affection.The second-order polynomial model was used to express the stachyose yield(Y)as a function of independent variables by the SAS RAREG program,the equation was shown as follows:Y=42.44+0.023X1+0.24X2+1.00X3+ 0.19X1X2-0.17X1X3+0.22X2X3-0.019X12-0.29X22-2.05X32.The optimum extraction conditions were as follows: extraction temperature 75 ?,extraction time 75 min,solvent-material ratio15:1,and the yield of stachyose was 42.26% under the process conditions.Studyed the effect of fermentation purification refining of stachyose by lactic acid bacteria,three kinds of yeast,Aspergillus oryzae and Schizophyllum commune.Lactic acid bacteria fermentation under the condition of 37?after 36 h,sucrose retention rate was 18.12%,glucose content increased,stachyose cleaved into manninotriose and retention rate was 65.89%,the purity of stachyose didn't improve.Three kinds yeast have the ability of rapid degradation of sucrose,the sucrose retention rate is less than 15% after 4h and almost completely consumed after 10 h.The yeast A and yeast C can degrade sucrose in the meanwhile consume monosaccharide from degradation of sucrose.But the yeast B can not,the fructose and glucose content increased.The yeast A,yeast B and yeast C fermentation after 4 h,stachyose retention rate was only 60%.Aspergillus oryzae fermentation after 22 h,stachyose partially degraded into manninotriose,the raffinose,sucrose,glucose and fructose were depleted,stachyose retention rate was 78.85% and accounted for about 83.34% of the total sugar.Schizophyllum commune fermentation after 48 h,sucrose retention rate was 15.27% and accounted for 1.07% of the total sugar;raffinose almost was no degradation and accounted for 4.86% of the total sugar content.The stachyose was slightly degradation,retention rate was 93.31% and accounted for 89.04% of the total sugar content.