Study On Extraction Of Active Ingredients And Preparation Of Functional Peptides From Whey Water Of Small Black Soybean

In this study, the active materials in small black bean whey were extracted; Whey protein were extracted and purificated,The optimum conditions of whey protein hydrolysis were find out,and then study of the antioxidant capacity of hydrolysis polypeptide in vitro;The functions of small black bean whey protein(SBWP) and small black bean whey peptides(SBWPE) in preventing alcohol-induced liver injury and acute Liver Injury were systematic studied through animal experiments. The results showed:1. The extraction of flavonoids and saponins:The result showed that extraction of saponins and flavonoids in the best conditions were:ethanol concentration of60%and70%, than1:30and1:40of material liquid ratio for2h, extraction temperature is70℃.under these conditions,the extraction rate of saponins was6.21%, of flavonoids was1.84%.2. According the determination, the PI of small black beans globulin was4.3; The highest content in the dry Crude extract of small black bean whey water was ash(37.84%),and then was total sugar (24.33%), and then was protein (17.81%); There were seven bands in SDS-PAGE electrophoresis clearly,the molecular weight of them were101.8kDa,62.4KD,61.8KD,25.3kDa,20.1kDa,18.4kDa,8.0kDa. Among them,the molecular weight of20.1kDa,which was trypsin inhibitor, had higher content.3.SBWP refined:flocculation, reduced sugar, microfiltration, ultrafiltration, vacuum drying, after these sequences, the purity of SBWP could reached more than70%.4.Use different proteases to hydrolysis SBWP under the most suitable condition of themselves. The results show that when the amount of bacterial acid protease enzyme was1500u/g protein and under the condition of pH=3,40℃to hydrolysis for4h, the effect was best, which peptide rate could up to91.153%; use bacterial acid protease to optimizate hydrolysis conditions, the best hydrolysis conditions were:pH=2, enzyme amount was2000u/g protein, hydrolyzed under45℃for4h. The total antioxidant capacity (0.835mg/mL) of SBWPE under the conditions was the highest;The tricine-SDS-PAGE electrophoresis results showed that the band of bacterial acid protease hydrolysis of the peptide distribution was relatively concentrated,80%were below1000Da.5. SBWP and SBWPE were fed to study the effect on preventing chronic alcohol-induced liver injury and CC14acute hepatic injury of mice. Results show that both SBWP and SBWPE could protect liver injury significantly.6.Skin moisture test showed that15%of SBWPE had better skin moisturizing effect, and it could significantly reduced the MDA content, improve the HYP content and CAT activity in the skin tissue, which suggested that SBWPE had the protective effect on the skin of long-term ultraviolet irradiated.