Induction Of Phytoalexins From Soybean And Kidney Bean And The Anti-tumor Activity Of Phaseollin

The family of phytoalexins play an important role in plant defense system, and it as functional foodingredient had received more attention.Phytoalexins had a variety of biological activities,such asantioxidation,estrogenic and estrogenic activity effect on human health and disease.In the present study,we selected soybeans and kidney beans as raw materials, and used analyticalHPLC, preparative HPLC, UPLC/MS and MTT as technologies.The aim of this study was to analyzeglyceollins induced by silver nitrate in soybeans,and compared the glyceollins production of differentvarieties of soybeans, as well as analyze the mechanism of glyceollins generation.On the other hand,thestudy on the phaseollin which was induced by oligosaccharides from kidney beans, and analyze thephaseollin on the growth inhibitory effect on breast cancer cells and uterine cancer cells. Theconclusions were as follows:1.The research focus on the production of glyceollins which was induced by10mM silvernitrate,and compared the glyceollins production of9different kinds of soybeans.The results showed thatglyceollins production was different for soybeans, green beans and black beans on the condition oftemperature of25℃, humidity of50%, and cultured in dark for3days. In this experiment, glyceollinsproduction from high to low were green beans, black beans and soybeans.The average glyceollinsproduction of3green beans was980μg/g dry bean weight,the average glyceollins production of3blackbeans was750μg/g dry bean weight, the average glyceollins production of3soybeans was470μg/g drybean weight.The content of glyceollins induced from9different beans as follow: Liuyuebai was1720μg/g drybean weight, Qingrenwu was1360μg/g dry bean weight, Daliqing was1030μg/g dry bean weight,Zhonghuang30was780μg/g dry bean weight, Peking was590μg/g dry bean weight, Zhongpin662was390μg/g dry bean weight, black soybean was310μg/g dry bean weight, Zhonghuang56was240μg/gdry bean weight, Guanqingdou was180μg/g dry bean weight.2. The experiment used Zhonghuang30and Qingrenwu as raw material to clarify the relationshipbetween glyceollins, daidzin and daidzein, and analyze the causes of glyceollin production ofQingrenwu higher than Zhonghuang30.As daidzein was the precursors of glyceollins synthesis, theresults showed that, on the one hand, because the content of daidzein in Qingrenwu was higher thanZhonghuang30, so it produced the content of glyceollins was higher.on the other hand, the daidzin inQingrenwu change to daidzein was stronger than Zhonghuang30on the condition induced by10mMsilver nitrate. The experiment proved that Qingrenwu was more suitable as raw materials for inducingthe production of glyceollins than Zhonghuang30.3. Application of chitosan oligosaccharide, pectin oligosaccharide, fiber oligosaccharide andalginic acid oligosaccharide as inducer, generated phaseolin from kidney beans. The contents ofphaseollin was determined by UPLC-MS/MS and HPLC. In the condition of5concentrations of oligosaccharide (1%,2%,4%,6%and8%), the result showed that for chitosan oligosaccharides,themost suitable concentration showing the highest level of phaseollin was4%,and phaseollinaccumulation was221.32μg/g.For alginic acid oligosaccharide,the most suitable concentration showingthe highest level of phaseollin was4%,and phaseollin accumulation was149.50μg/g.For pecticoligosaccharides,the most suitable concentration showing the highest level of phaseollin was6%,andphaseollin accumulation was265.86μg/g. For fiber oligosaccharide,the most suitable concentrationshowing the highest level of phaseollin was8%,and phaseollin accumulation was211.30μg/g.4.The research focus on phaseollin as phytoalexins possessed both estrogenic and antiestrogenicactivities on breast cells and uterine cancer cells.The experiment used Cal-51, Caski, Hela and Mcf-7asmaterials.The results showed that phaseolin had obvious inhibiting effect on the growth of4kinds ofcells. On the condition of50ug/ml phaseollin, the inhibitory effect from strong to weak were Cal-51was93.01%，Caski was89.68%，Hela was81.90%，Mcf-7was66.49%. In addition, the IC50of phaseolinon Hela and Mcf-7were respectively36.31μg/ml and46.73μg/ml.