Isolation Of Antioxidant Peptides And Its Effect On Carbofuran Damaged AGS Cells

In order to ensure food production,a large number of agricultural use carbamate pesticides to remove pests and weeds,and carbamate pesticide residues in the soil,water and relay into the human body,resulting in oxidative damage to human tissues and organs,The formation of teratogenicity,carcinogenicity.In this paper,we study the casein hydrolyzate formed by the enzymatic hydrolysis of natural casein,and the casein hydrolyzate reacts with plastein reaction protein to produce new antioxidant activity peptide.The antioxidant activity of plastein reaction product is related to casein hydrolyzate compared with a higher antioxidant activity.The plastein reaction products with high antioxidant activity were separated by Sephadex G-15,and the isolated products were collected and the antioxidant activity of the different components was determined.The protein isolates with the highest antioxidant activity and the highest antioxidant activity were treated with carbamylamine-induced AGS cells.The cell viability was detected by CCK-8 method,the changes of GSH content and the change of ROS content to study the effect of antioxidant peptides on cells damaged by carbofuran.The results are as follows:(1)Hydrolysis of casein by papain to obtain casein hydrolyzate.Through the plastein reaction,the casein hydrolyzate was introduced into the foreign amino acid to obtain the modified product.The results showed that the antioxidant activity of plastein reaction product 3 at 5 h was relatively high,which could be used as the next step for separation(2)The protein-like modification product(5 h modified product 3)was separated by dextran gel.The optimum separation efficiency was obtained by different particle size,different flow rate and different loading.The Sephadex G-15 was used at a flow rate of 0.5 mL/min and the loading was 20 mg/mL.The best separation effect was got.(3)Preparation of antioxidant peptides of high purity antioxidant peptides on the basis of the best conditions of separation.The antioxidant activity of the products of peak 2(P2)was compared with that of plastein reaction products(TFA)and methanol(TFA)were used as the mobile phases to determine the purity of the products.The results showed that the products of the two plastein reaction products 3 had a significant increase in antioxidant activity(P<0.05),The results showed a single peak,indicating high purity of antioxidant peptide.(4)The damage of AGS cells was induced by carbofuran,and a stable cell injury model was established.Plastein reaction products and the isolated products could significantly increase the cell viability,decrease the level of ROS and increase the content of GSH,thus the effect of the products could be achieved by the method of casein hydrolyzate and plastein reaction products.It was proved that the antioxidant peptides with different purity could reduce oxidation of AGS cells from damage induced by carbofuran.In summary,the plastein reaction product of casein hydrolyzate was isolated to obtain the isolated product,in which P2 had the highest antioxidant activity and the strongest effect on carbamine-induced AGS cells.