Study On The Preparing Isoflavoues Aglycone By Composite Biological Method

Soy isoflavones powder of commercially available as a sample,Increased the percent conversion of Isoflavoues Aglycone by Composite Biological method then complex biological process conditions were optimized.Frying,microwave and oven heating were used to preprocess the soy isoflavones materials commercially.The influence on the absolute quality of Isoflavoues Aglycone which was conversed enzymatic soy isoflavones was discussed through the preprocess method.The results showed that:frying was significantly better than the oven and microwave method(p<0.05),the absolute quality of fried Isoflavoues Aglycone was higher than the oven,microwave 1.238,1.240mg/g diferently.Pretreatment conditions were optimized using single factor and response surface methodology to obtain results:flying time 107.68 s,frying temperature 131.29 ℃,the amount of material 13 g that Isoflavoues Aglycone conversion rate was 71.4%.The corrected value were frying time 108 s,frying temperature 132℃ and the amount of material 13 g that Isoflavoues Aglycone conversion rate was 72.23%.The Aspergillus niger β-glucosidase as a research object,entrapped immobilization method was used,the enzyme activity recovery rate was evaluation index.Sodium alginate,chitosan and sodium alginate-chitosan chitosan were selected as the carriers,the effects of the entrapped immobilization was compared.The Results showed that sodium alginate chitosan was the best supporter.Immobilization conditions of single factor and response surface experiment were taked,the optimal conditions were obtained:the alginatemass fraction of the mixed carriers was 1.91%,the chitosanmass fraction of the mixed carriers was 1.94%,rhe temperature was 40.20 ℃,the crosslinking time was 0.98h.At this time,the enzyme activity was 112400IU/mL,the recovery of the enzyme activity was 66.18%.After the three recovery experiments the recoveries of the enzyme activity were 65.61%,64.48%and 64.16%.β-glucosidase producing-capacity in LJ-G1,LJ-Q2 of vegetarian intestinal bacteria compared to Aspergillus niger with Nitrophenol-β-D-glucoside(pNPG).Then β-glucosidase producing-condition was optimizated by a single factor and response.surface test,the results showed:β-glucosidase could be produced by LJ-G1,LJ-Q2,enzyme producing-ability of LJQ2 was higher than LJ-G1.enzyme producing-ability of LJ-G1,LJ-Q2 was higher than Aspergillus niger in 64 h fermentation.Optimal conditions for enzyme production were that medium was pH 8.04,culture temperature was 38.31℃,culture time was 38.21 h.Validate test was under the optimum conditions,the enzyme activity reached 1.7 IU/mL.conversion of Soybena Isoflavones Aglycon was 40%in optimum condition.Pretreatment soy isoflavones by recombination fermentation of immobilized enzymes and bacteria were used then detected the absolute quality of the soy isoflavones by HPLC.The condition of recombination fermentation was optimizated by a single factor and response surface test,the results showed:temperature of recombination fermentation was 54.03℃,time of recombination fermentation was 3h,the initial ph was 7,enzyme dosage was 7.29%.Through the proven test,the absolute quality of soy isoflavone aglycone was 13.76 mg,the conversion rate of soy isoflavone aglycone was 76.8%.Macroporous resin purified fermentation broth then detected the absolute quality of the soy isoflavones.The condition of purifying was optimizated by a single factor and response surface test,the results showed:solid-liquid ratio was 1.82:1,pH was 4.01,resolution time was 6.03 h,through the proven test,obtained the absolute quality of soy isoflavone aglycone was 3.45mg,analysis rate was 33.5%.Then freeze-dried to prepare soy isoflavones powder of which purity of soy isoflavone aglycone was 47.3%.