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Preparation of Scallop Oxidation Active Peptide and the Activity Research

Author: ZhangLiLi
Tutor: WangJunHu
School: Harbin Institute of Technology
Course: Genetics
Keywords: Chlamys farreri Antioxidant activity peptide Enzyme hydrolysis Separation and purification C. elegans lifespan
CLC: TS254.1
Type: Master's thesis
Year: 2013
Downloads: 102
Quote: 0
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Abstract


Oxidation is one of the main reasons for human disease and pathogenesis. Suchas free radicals attack protein, lipid, and nucleotides to cause cell damage andapoptosis, which can lead to tissue damage, cause the body’s aging, and induce themalignant diseases such as cancer, etc. In essence, all the cell components andspecial ingredients are vulnerable to be influenced by free radicals. It has beendemonstrated that antioxidant peptides can scavenge free radicals, chelate metal ions,and so on by large number of studies. And Scallop is one of the important marineshellfish resources produced in large quantities from north to south throughout thecoastal area in our country. Not only rich in nutrition, delicious, but also hygienicalfunction. So Chlamys farreri was chosen for the research, Chlamys farreriantioxidant active peptide was prepared by enzyme hydrolysis and its effects on thelifespan of C. elegans was further studied.Papain, flavourzyme and trypsin were selected to hydrolyze Chlamys farreriprotein through sole enzymolysis, compound enzymolysis and multi-stepenzymolysis technology and compared by hydroxyl removal capacity and peptideyield degree of hydrolysis. Consequently, trypsin digestion of the sole enzymolysishas been determined as the optimal digestion way. Chlamys farreri protein digestionconditions were optimized by Orthogonal design assistant IIV3.1. Under thisoptimized conditions, the hydrolysis of Chlamys farreri protein’s hydroxyl removalcapacity (SA) was91.09%, median inhibitory concentration (SC50) was35%, peptideyield was56.69%, degree of hydrolysis (DH) was73.25%.High purity of antioxidant activity of peptide was obtained by ultrafiltration,molecular sieve chromatography (SEC, Superdex Peptide10/300GL) and highperformance liquid chromatography (HPLC, Zorbax300SB-C184.6×2505μm). itsmolecular weight distributes in0.1-5kd, and its predicted chain length is4-14aminoacids.Select C. elegans as the research object of longevity experiment, by feedingwith three different concentrations (0.1%,1%and10%separately) of the hydrolysisof Chlamys farreri protein and higher anoxidation activity of Chlamys farreri proteinpeptide obtained by purification (<10kd), while compared with E.coli OP50ascontrol group; Chlamys farreri protein antioxidant activity peptide (<10kd) has moresignificant effect on lifespan of C. elegans by observing and recording the changes ofphysiological characteristics (lifespan, activity and reproductive capacity, etc.)during its life and by using the SPSS software to deal with the data.

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