The Study On The Activity Of Isoflavones In Fermenting Soybean

In this experiment,the fermenting soyfood —tempe had been studied,and its activity of isoflavones,including the activities of anti-oxidation,anti-fungus and the inhibitation to the growth of cancer cells . The fermenting measure of tempe could be definited initially as that soaking soybean for 18²4h—steaming soybean for 20min by 0.08Mp—mixing 0.8% lactic acid—suppling fungal inoculum—fermenting at 36(±1)℃for 30h. The extraction of isoflavones:the tempe would be degreased by n-Hexane,added the ethanol ,circumfluence and extration, then decompressed and concentrated the liquid to the dry.This isoflavones in mix would be extracted by E.a, uniting the extration liquid . the E.a would be vaporized at the station of decompression.then got the commix of isoflavones,used it to the study of anti-oxidation,anti-fungus. The component would be analysed by HPLC. The result was :the change of the gross was not obvious of isoflavones in former and later,but the content of aglucones in tempe had been increased, the concentration of aglycon mol increased from 4.42×10^-7 mol/gto 30.9×10^-7 mol/g, the later was the 7 times to the the former,but the glucoside mol had decreased from 64.9mol/gto 48.2 mol/gwhose content was the 74.27% of the former. The isoflavones would be pured throμgh D4006 macroporous resins,and determined the content of isoflavones by UV-751 spectrophotometer,the substance of pured was confirmed as the isoflavones with UV—7500UV scanner, the purity of the isoflavones could reach 35% or so by this method. The mensuration of the activities of anti-oxidation of isoflavones: the animal fat was used as the experimental material,then measure the POV and TBA at the specified period. The method of Marklund would be used to determined the scavenging rates for the O₂^-·.The result was found:,POV was 30.8 ,38.1 and 162.7 adding tempe isoflavone ,soybean isoflavone and the contrast in lards sample on 21 days. The result of TBA was similar with the POV.Tempe isoflavones in 100 ²5times for O₂^-·.svaven lead as 45. 6%²6. 8%, the scavenging rates which was contrasted the sample were 43.1%²4%. The result indicated that the tempe was better in anti-oxidizeing activation than soybean isoflavones. The experiment of anti-fungus:one is the inhibitation experiment of Staphylococcus aureus, fermenting the soybean,at the same time,inoculated Staphylococcus aureus,separated and checked the fungus by the general means after the fermentation. The result was found: both soybean and tempe had the ability of inhibitation to bacilli in different degree, the tempe fermented 30h could not be checked the lively bacilli,but the soybean fermented 42h,the content of bacilli had reached 5.2×108 ufc/g.The effect of inhibitation of fermented soybean was better than the unfermented soybean.The other is the method of general filter paper to determine the circle of the inhibitation-fungus.The effect of inhibitaton differed with the different fungus:the diameter of the circle of the inhibitation-fungus of tempe isoflavones was 35.5mm,33.5mm,16mm for E.coli, Salmonella typhimurium and Staphylococcus aureus, but the soybean isoflavones was 20 mm,26.5 mm,13.5 mm,the effect of inhibitation for yeast and the milddew. was not obvious. MTT colorimetry was used to determine the inhibitation of tempe and soybean isoflavones extraction whose purity was 35% for the Hela,SP2/0 and CEF cells, calculated the inhibitation ratio. The result was found: the inhibitation ratio for the SP2/0 was 96.9%;for the Hela was 69.5% when the concentration of tempe isoflavones was 20μg/ml, the inhibitation ratio for the SP2/0 was 83.6%;for the Hela was 60.5% when the concentration of soybean isoflavones was 20μg/ml.With the reducement of concentration,the inhibitation would depress. at 1.25μg/ml, the effect of both tempe and soybean isoflavones did not show the inhibitation for the SP2/0. the inhibitation ratio for the Hela is 34.3% and 23.4%. The result indicated that the inhibitation of this two isoflavones for the CEF was weak, however,for the cancer cells was strong,the inbihitation of isoflavones in tempe for the cancer cells was strong obviously.