Studies On The Preparation Of Peptides By Enzymatic Hydrolysis From Sheep Spleen And Their Antioxidation Activity In Vitro

In the present research papain and trypsin were used to hydrolyze spleen protein and then the antioxidant properties of the hydrolysis were analyzed. It was done to abtain the optimal condition of hydrolysis of spleen protein. The optimum hydrolysis parameters were: the enzyme application accounted for 5.0%(enzyme/substrate),reaction conditions of pH 7.0, temperature 60#, time 5.5 hours and the degree of hydrolysis being 60.59% with stronger antioxidative activities. The hydrolysate was fractionated by gel filtration on a Sephadex G-25 column.Four fractions(their serial number is F1, F2, F3, F4) were gained,and then their antioxidation activity were tested by DPPH . The result showed that F3 was the most active antioxidant compared with the other fractions .The fraction was analyzed by Tricine-SDS-PAGE ans the molecular weight was estimated by this method.Experiments about the inhibitory effect of the peptides FS on lipoxygenase, the auto-oxidizing of lipid were performed . It turned out : In different free radical generating systems: F3 was able to inhibit the OH effectively with the larger the concentration; Scavenging effects on 1,1-Diphenyl-2-picryl-hydrazyl(DPPH), the antioxidative activities of F3 were stronger with the increasing concentrations; However, FS wasnot effectively eliminated O2-. A Significant(p < 0.01) synergistic effects of citric acid plus spleen polypeptides were obtained by DPPH method .however no significant synergistic effects when employing assorbic acid plus spleen polypeptides; In the systems of swinelard and phosphatidylcholine liposome, it was found Fa had some extend of antioxidant activities, the larger the concentration, the stronger the antiooxidation activity.