Preparation And Antioxidant Activity Research Of Peptide From Apostichopus Japonnicus Intestinal Tract Tissue

Intestinal tissue was byproducts of processed Apostichopus japonicus, which were selected as materials to determinate the basic nutrition firstly, and then sole factor test and response surface test analysis were chosen to optimize the technology of hydrolysis of the protease of papain, protease of compound and, protease of flavor. Hydrolysates were graded by ultrafiltration membranes with various MWCO(molecular weights of cut-off).Chemical antioxidant activity and cell protection of cascade peptide were studied. Finally, yield, activity and ability of protection were selected as directions and purpose for peptide separation and purification.Then as follows was the main research content:1. Nutrients detection and inspection of the Apostichopus japonicus intestinal tissueAs byproducts of processed Apostichopus japonicus intestinal tissue, which nutrients were adundanted and balanced, 63.056g/100 g crude protein content.Such a many protein content, so that it could be used as raw material for preparing the peptide.Optimization of the Enzymatic Hydrolysis Condition of Apostichopus japonicus Intestinal tissueFirstly, considered the product feasibility, fresh factors and safety to choose appropriate protease, optimized the enzyme hydrolysis condition by considering the temperature of enzyme solution, the reaction time and the concentration of each other protease. Then, determined the optimal process by the used of the response surface method. Finally, we determined the optimal conditions of hydrolysis of papain enzymatic as follows: with the enzyme dosage 26 390 u, emperature 50.10 ℃,enzymatic hydrolyzed 2.84 h, the degree of hydrolysis reached 41.82%.This experiment adopted exogenous neutral enzymes, included there kinds,coupledwithautolyticenzymesinapostichopusjaponicusintestinaltracttissue,puttedupavarietyofenzymatichydrolysissystem.theresultsofthisexperimentshowedthattheconditionsofoptimizationofenzymaticcatalyzedhydrolysisisreasonableandeffective.basedonthesecurity,itnotonlyimprovedthedegreeofhydrolysis,butalsothehydrolysatewhichwasfresh,brightcolor,deliciousclear,anduniform,whichcouldbeusedasauniqueflavortodevelopfunctionalfoods.3.isolationandpreparationandactiveresearchofcascadepeptidethisexperiment,gradientdilutionmethodwereusedtooptimizatetheultrafiltration,whichusedtoseparatecascadepeptidefromenzymesolutionofapostichopusjaponicusintestinaltissue.theresultsshowedthatafterfourultrafiltration,peptideswhichmolecularweightlessthanthefilmpackagehadbeenintotheliquidfiltration.sothat,itachievedthepurposeoftheseparationofpeptides.fourcascadepeptideswithdifferentmolecularweightrangewereobtainedbytheultrafiltrationseparation,whichwenamedt3withmolecularweightlessthan650 u,t2withmolecularweight650-1000 u,t2withmolecularweight650-1000,t1withmolecularweight1000-10000 u,t0withmolecularweightmorethan10000u.afterdetectedtheyieldsoffourcascadepeptideswere95.32%、2.57%、1.70%、0.41%。theresultsofthechemicaloxidationresistanceoft1,t2andt3showedthatthet1,t2andt3wererevealedastrongabilitytoremovingdpph·,superoxideanionandhydroxylfreeradical,withinacertainrange,ithadacertainscavengingeffectinadose-dependentmannermeannedthatwerepositivelyrelatedtothesampleconcentration.inwhich,theabilitytoremovedpph·andsuperoxideanionoft3wasbetterthant1andt2.atthesametime,almosttheabilitytoremovehydroxylfreeradicaloft2andt3,whichbetterthant1.isolatedcascadedpeptidesofjaponicuswhichdetectedtheabilitytoprotecttheprimarymacrophagesfromthedamageofhydrogenperoxide.beenestablishedthroughinjuryofhydrogenperoxideofprimarymacrophagemodel,selectedtheconcentrationofh2o2was160μmol/l.detectedtheabilityofprotectiveeffectsofeachsampleofcascadepeptidesbytheuseofmtt.theresultsshowedthatthesampleofeachcascadepeptideshadasignificantcelldamageprotection,theprotectiveeffectoft3 peptidewhichisbetter.4.purificationandphysicochemicalanalysisofapostichopusjaponnicusintestinaltracttissuethis experimentusedchromatographyofmacroporousadsorptiveresinstoseparatet3forfurtherseparationandpurification.andthendetectedtheantioxidantofisolatedcomponents,whichincloudedtheabilitytoresistsuperoxideanionanddpph·.thefivecomponentsoft3 separatedthroughchromatographyofmacroporousresin,whichnamedoff1,f2,f3,f4andf5,andeachcontentswere5.657%,29.032%,16.543%,32.862%and15.682%.bymeasuredtheyieldoffreeradicalscavengingabilityandcomprehensiveanalysis,usedf2 asafurtherseparationandpurification.this experimentusedhsccctoseparatef2,whichsubjectedfromchromatographyofmacroporousadsorptive,forfurtherseparationandpurification.byselectingthepropersolventsystems(methyltert-butylether,n-butanol:acetonitrile:1%trifluoroaceticacid=2:1:1:5),obtainedfourpeaksoff2 peptide,whichnamedf2-1、...