| Chlorella vulgaris is a kind of spherical or elliptical unicellular freshwater algae, 3 to 8 microns in diameter, asexual reproduction, is one of the earliest lives on earth which appeared in more than 20 billion years ago. Chlorella is highly efficient photosynthetic plants can autotrophic growth and reproduction. Because of its wide distribution, and extremely easy to obtain, it can be a kind of marine biological resources with high protein, but the practical value has not yet been developed. At present, few studies on C hlorella peptide about control diabetes, so it need to be further studied. There is a correlation between the antioxidant activity of Chlorella peptide and the treatment of diabetes mellitus. In this paper, Chlorella is used as a raw material for preparing antioxidation polypeptide, and then, explore the antioxidant and hypoglycemic effect of Chlorella peptide through the analysis of the lifespan, enzyme activity, oxidation cluster level in vivo of C.elegans to establish a theoretical foundation for subsequent trials.This paper studied on C hlorella breaking conditions in order to avoid the loss of protein by Cell wall broken. According to the single factor experiment and the orthogonal experiment results, four best C hlorella breaking conditions were confirmed: the oxidation cluster leve l3.9% the concentration of sodium hydroxide, extraction time of 26.8 min, extraction temperature of 47.3oC, the ratio of material to liquid 1: 31, under these conditions, the extraction rate of soluble protein was 5.20%. The method is simple, does not involve large instrument operation, extraction time is moderate but with high extraction rate of soluble protein.Flavourzyme is used as a protease to prepare peptide according to the results of single factor experiment. Under the condition of 50 oC, p H6 for 5 hours with flavourzyme, the soluble protein is hydrolyzed into peptide which has a high free radical scavenging rate. The IC50 of DPPH, superoxide radical and hydroxyl radical is 0.325mg/m L、0.325mg/m L and 0.139 mg/m L. Peptide can extend the lifespan of C.elegans 39.20%, meanwhile, has a significant extension of lifespan of C.elegans in high glucose environment. 77.87% and 1575.21% are upgraded on superoxide dismutase and catalase activity, and reduce ROS 13.66%. The IC50 of α-glucosidase activity, α-amylase activity, invertase activity is 1157.81 mg/m L、1.70 mg/m L and 3.38 mg/m L, extend the lifespan of C.elegans in the high glucose environment. The peptide solution purified with two different molecular weight sephadex gel, get two kinds of polypeptides with similar molecular weight about 11.2 k Da. Experiments show that, peptide with antioxidant ability can inspire the oxide hydrolase activity within the cell, especially for the promotion of catalase activity, its mechanism needs further study. Based on the elimination of excess free radicals in cell, the peptide can extend the it’s life. Polypeptides exhibit higher polysaccharide(disaccharide) enzyme inhibitory activity, can depress the polysaccharides activity, reduce the concentration of glucose, to control the absorption of glucose. Peptide Polypeptide has obvious promotion on the average lifespan of C.elegans in high glucose environment, C.elegans in high glucose environment has higher oxidation level similar to pancreatic islet β-cell, therefore, the polypeptides can protect islet β-cell cells in diabetes patients in the theory.In this paper, polypeptides have good in vitro and in vivo antioxidant capacity, at the same time showed higher hypoglycemic effect, provide the basis for the development of new anti diabetes drugs. |
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