| In this study,Mung bean was used as raw material,and the ACE inhibitory peptide in mung bean was extracted by ultrasonic-assisted enzymatic method.SephadexG-25 and SephadexG-15column chromatography were used to separate and purify the hydrolyzed product,and theseparation conditions were determined.The in vitro inhibition rate was determined by the three fractions purified by SephadexG-25 and the four fractions purified by SephadexG-15,and then subjected to in vitro antioxidant test on each fraction.The in vitro test results showed that mung bean ACE inhibitory peptides had certain scavenging effects on DPPH,ABTS,superoxide anion,and hydroxyl radicals.The aging model of mice was established with D-galactose,and different doses of mung bean polypeptides were given.The contents of MDA,SOD and GSH-Px inserum and liver of mice were determined.Through the gavage treatment of mice with different doses of mung bean polypeptides,the acid phosphatase content,alkaline phosphatase content,lysozyme content,half serum hemolysis value,lactate dehydrogenase content,clearing carbon clearance ability and other indicators were measuredto determine the immunomodulatory capacity of mung bean.The main findings of this experiment are as follows:?1?Purification of the extracted mung bean peptides showed that the optimum conditions for separation of SephadexG-25 were as follows:the eluent was deionized water and the sample concentration was 9 mg/mL(loading volume was 2 mL.The elution flow rate was 0.7 mL/min.Results:Three components were obtained and the inhibitory rate of these three components was determined in vitro.The inhibition rate of the V3 component was 93.67±3.43%,which is the highest among these three components.Then,the highest inhibitory rate of the V3 component was purified again.The best separation conditions were obtained by using the SephadexG-15through the above steps:The elution agent was deionized water and the sample concentration was 20 mg/mL?loading volume 2 mL?,elution flow rate was 0.8 mL/min,and the in vitro inhibition rate of these components was also determined.The inhibition rate of L3 component was the highest,reaching 95.89±2.26%.?2?F1 component was further purified by RP-HPLC using the collected L3 fraction and the ACE inhibition rate was 97.12±1.47%.?3?All of the purified components were subjected to in vitro antioxidant tests.DPPH,ABTS,superoxide anion,and hydroxyl radical scavenging rates were used as evaluation indicators.Among them,the antioxidant effect of V3 and L3 components is the best.The in vitro test results showed that when the concentration of mungbean polypeptide reached 5mg/mL,the V3 component had a hydroxyl radical scavenging rate of 57.26%,the superoxide anion clearance rate was 54.54%,the DPPH radical scavenging rate was 64.79%,ABTS free The clearance rate of the base is 94.33%.?4?The in vivo antioxidant test was performed.The results showed that the content of MDA in serum and liver of mice showed a downward trend,and the activity of SOD and the activity of GSH-Px showed an upward trend.Among them,compared with the model group,the liver MDA content of mice in the high-dose group decreased by approximately 30.16%,and the SOD activity in the serum increased by approximately 14.03%,which was similar to that of the positive?VC?group.The GSH-Px activity in the high-dose group was increased by about 66.59%compared with the model group.?5?In vivo immune indicators were measured.The results showed that the content of acid phosphatase was significantly correlated with the blank group?P<0.05?,and the high dose group reached 13.60±0.28 U/L,while other indicators showed a certain dose effect,indicating that mung bean polypeptide had certain immunoregulatory function. |
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