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Preliminary Studies on Microbial Conversion of Isoflavones

Author: MiaoHui
Tutor: ZhaoHai
School: Chengdu Institute of Biology
Course: Botany
Keywords: Genistein Daidzein Enzymatic β - glucosidase
CLC: Q939.97
Type: Master's thesis
Year: 2005
Downloads: 276
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Abstract


In this paper, the extraction and separation of daidzein and genistein, and quantitative methods. Soybean meal defatted, defatted soybean meal first with n-hexane and then 70% ethanol extraction, and concentrated ethanol extract to alcohol-free, the same volume of ethyl acetate and then with a water phase extracted three times were combined and ethyl acetate phase was concentrated under reduced pressure. GF 254 chromatography plate, toluene - chloroform - acetone (4:1.8:1.8) eluent ethyl acetate phase thin-layer chromatography, determined through experiments R f = 0.50 substance genistein daidzein, the R f = 0.63 substance by UV - visible spectrophotometry quantitative. Under our experimental conditions, the relative standard deviation of daidzein and genistein extraction rate were 1.40% and 0.87% of the content of the relative deviation of the measured values ??of 3.69% and 3.52%, respectively. This indicates that the extraction and separation of daidzein and genistein The established method is stable and effective, accurate and reliable quantitative method. By judging the extent of whether or not hydrolyzed or hydrolyzed soy isoflavone glucosides to filter the soybean isoflavone glycoside decomposing bacteria, to be measured before and after hydrolysis aglycones content changes, the entire process is more complicated. R f = 0.82 Chu found in the thin layer chromatography samples daidzein and genistein outside, the third fluorescence spot, the HPLC-PDA, mass spectrometry and the color reaction analysis to determine soybean fine alcohol, and confirm the presence or absence of fluorescence intensity of the soybean refined alcohol can be used as to determine whether soy isoflavone glucosides hydrolysis or hydrolysis sign greatly simplified screening soybean isoflavone glucosides decomposing bacteria process. Also studied the separated and purified higher rates of hydrolysis of soybean isoflavone glucosides the black Aspergillus N 3 soy isoflavone glucosides, p-NPG and geniposide hydrolysis situation by orthogonal experimentally determined the black Aspergillus N 3 on of genistin the best hydrolysis conditions for temperature = 52 ° C, time = 12h, pH = 4.5, Ca 2 concentration = 30 mmol · L -1 , was 41.42% enzymatic hydrolysis conditions on of daidzin the best temperature = 52 ° C, time = 12h, pH = 5.0 Ca 2 concentration = 60 mmol · L -1 , enzymatic hydrolysis rate of 95.71%. N 3 the The Aspergillus enzyme production enzymolysis p-NPG good effect, but only 30.82% of the enzymatic hydrolysis rate gardenoside. The the aspergillus niger N 3 pairs the different glycosidic exhibit different enzymatic effect, this may be caused by the structural differences of the different glycoside.

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