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1、桃仁多肽的酶法制備及其抗氧化活性分析楊玉蓉1李安平1,*鐘政昌2李剛1(中南林業(yè)科技大學(xué)食品科學(xué)與工程學(xué)院1,長(zhǎng)沙410004)(西藏農(nóng)牧學(xué)院食品科學(xué)學(xué)院2,林芝860000))摘要:以桃仁蛋白為原料,對(duì)響應(yīng)面法優(yōu)化多肽制備條件和酶解多肽各組分的抗氧化活性進(jìn)行了研究。結(jié)果表明:以底物濃度3.5%、pH10.4、加酶量4200U/g、酶解時(shí)間4.9h條件下進(jìn)行堿性蛋白酶酶解,可獲得桃仁蛋白的最高水解度(61.12±0.7)%和多肽得率(67.91±0.5)%。桃仁蛋白及其酶解物的SDS-PAGE凝膠電泳圖表明,酶解后的多肽分子質(zhì)量大部分小于12000u。酶解液
2、(PPH)及其超濾分離所得4組分分別采用DPPH自由基清除能力、ABTS自由基清除能力、亞硝酸根離子清除能力和總抗氧化能力等4個(gè)體外抗氧化活性測(cè)試,結(jié)果顯示不同分子質(zhì)量組分間的抗氧化活性存在顯著性差異(P﹤0.05),分子質(zhì)量越小,其抗氧活性越強(qiáng)。RP-HPLC分析表明,抗氧化活性最強(qiáng)的P-IV組主要由9種具有一定疏水性的多肽組成。關(guān)鍵詞:桃仁多肽酶解抗氧化活性SDS-PAGE反相高效液相色譜中圖分類號(hào):TS209文獻(xiàn)標(biāo)志碼:A文章編號(hào):PreparationofPeptidesfromPeachKernelProteinbyEnzymolysisandIt
3、sAntioxidantCapacityinvitroYangYurong1LIAnping1*ZhongZhengchang2LiGang1(CollegeofFoodScienceandEngineering,CentralSouthUniversityofForestryandTechnology1.Changsha410004)(CollegeofFoodScience,XizangAgricultureandAnimalHusbandryCollege2.Linzhi860000)Abstract:Thestudyaimstooptimizethe
4、enzymatichydrolysisconditionanddetectinvitroantioxidantcapacitiesofhydrolyticPeachkernelpeptides.Thedegreeofhydrolysis(DH)andextractionrateofpeptide,asresponses,wereoptimizedbyusingResponseSurfaceMethodology(RSM).Theoptimizationresultsweresubstrateconcentrationof3.5%,pHvalue10.4,en
5、zymedosage4200U/gandhydrolysistimeof4.9h.Underthisoptimumcondition,theDHandextractionrateofpeptidewere(61.12±0.7)%and(67.91±0.5)%,respectively.TheSDS-PAGEanalysisshowedthatthemolecularweightofpeachkernelhydrolyticpeptideswasunder12000u.4componentswithdifferentmolecularweightweresep
6、aratedfrompeachkernelproteinhydrolysate(PPH)byultra-filtration.ThescavengingcapacitiesofDPPHradical,ABTSradicalandnitriteaswellastotalantioxidantcapacitytestwereadoptedtoevaluatetheantioxidantcapacityof4componentsandPPH.Resultssuggestedthattherewasasignificantdifferenceinantioxidan
7、tcapacitybetweendifferentmolecularweightcomponents(P﹤0.05).Thesmallerthemolecularweight,thestrongertheantioxidantactivityis.Finally,thehighestantioxidantcapacitypeptideP-IV(﹤3000u)wasanalyzedbyReversedPhaseHighPerformanceLiquidChromatography(RP-HPLC).TheReversedPhaseHighPerformance
8、LiquidChromatogramofP-IVin