果胶酶对覆盆子和黑加仑果汁饮料产量及其花青素可萃取性的影响

Effectofpectinolyticjuiceproductionontheextractabilityandfateofbilberryandblackcurrantanthocyanins果胶酶对覆盆子和黑加仑果汁饮料产量及其花青素可萃取性的影响汇报内容IntroductionMaterialsandmethodsResultsanddiscussionConclusionAbstractBilberriesandblackcurrantsdarkblueberriesrichinanthocyanins,wereprocessedwithanaidofcommercialpectinolyticenzymepreparations,andtheeffectofprocessingonberryanthocyaninswasinvestigated.Theenzymepreparationsweredosedbasedontheirpolygalacturonaseactivityfrom1to100nkat/gofberrymash.Thejuiceyieldsweredeterminedbyweighing,andanthocyaninanalyseswereperformedwithHPLC.Thebilberryandblackcurrantjuiceyieldsincreasedsignificantlyinenzyme-aidedtreatmentswithcomparisontocontrol,evenwiththelowest(1nkat/g)polygalacturonasedosage.Theanthocyaninyieldincreasedbyupto83%forbilberriesandupto58%forblackcurrantsinenzymeaidedtreatmentsascomparedtocontrol.Theresultsshowedthathigherpolygalacturonasedosagewasneededforblackcurranttoachievethemaximaljuiceandanthocyaninyieldsthanforbilberries.Thestabilityandtheprofileofextractedanthocyaninsweregreatlyaffectedbytheglycosidasesideactivitiespresentintheenzymepreparations,whichwereabletohydrolyzecertainanthocyaninstothecorrespondingaglycones.Inaddition,thedataindicatethatanthocyanidinrutinosidesweremoreeasilyextractedthanthoseofglucosides,whichprevailedoverthearabinosidesandgalactosides.Thus,priortoprocessingitisimportanttoknowtheintactanthocyaninstructuresoftherawmaterial,andtheactivityprofileoftheenzymepreparationtoobtainoptimalanthocyaninextractabilityandenzymedosage.摘要富含花青素的覆盆子及黑加仑果在通过添加某种商业果胶酶制剂进行处理后，对其中花青素所受的影响进行调查分析。酶制剂将根据它们自身的聚半乳糖醛酸酶活性从1-100nkat/g分别添加加入到果泥中。重量决定果汁的产量，对花青素的分析将被高效色谱分析法所呈现。最低剂量的聚半乳糖醛酸酶试剂（1nkat/g）都可以使覆盆子和黑加仑果汁的产量相比于空白组来说有明显的增加。经过酶制剂处理的覆盆子和黑加仑果中的花青素产量分别提高到了83%和58%。结果表明，为了获取最大的果汁与花青素产量，黑加仑所需的酶制剂剂量要多于覆盆子。花青素提取的标准和简况很大程度的被酶制剂中糖苷酶的活动所影响，这些酶的活动可以将某些花青素水解为所对应的糖苷配基。另外，数据表明芸香糖苷相比于葡萄糖苷、阿拉伯糖苷和半乳糖苷而言更容易被提取。因此，在处理之前知道原料中完整的花青素结构和酶活动的基本规律从而过得最佳的花青素提取量还有酶的添加剂量是很关键的。Introduction黄酮类物质（Flavonoid）是一群来自于水果、蔬菜、茶、葡萄酒、种子或是植物根的化合物，属于生理活性物质。黄酮类在许多植物和蔬菜中都有发现。在西方的日程饮食中，它们的日均消费量约为1克。有大约超过4000种复合物属于黄酮类，它们被分为5个亚类：黄酮、黄埦醇、总黄酮、黄酮醇、和花青盐。黄酮类具有抗氧化、抗癌、抗血管增生、消炎、抗变应性、和抗病毒的功效。花青素是一种植物的次生代谢产物，其属于类黄酮类物质，存在着典型的环苯甲酰和羟基肉桂的结构。超过4000种植物类黄酮已鉴定,其中大约600个拥有天然花青素结构。花青素一般较多的存在在红和蓝色的水果与花朵中，因此它们常常被当作食物的着色剂使用。另外，花青素拥有很强的生物活性，它们的抗氧化性、消除自由基能力、抗诱变性、抗炎性与抗癌性经过研究后，对人类的健康很有益处。果胶酶常用在工业浆果的加工中促进果汁的提取。它可以对植物细胞壁进行破坏使汁液流出，大大增加果汁的产量。覆盆子和黑加仑果中富含着花青素，本文研究酶制剂对覆盆子和黑加仑果中花青素提取的影响，目的在于使用最少剂量的酶，产生最小的负面效果，以获得最大的花青素提取率。Materialsandmethods酶制剂：EconaseCE、BiopectinaseCCM、PectinexSmashXXL、PectinexBE-3LJuiceprocessingThedetailsofalaboratory-scalejuiceproductionhavebeenpresentedearlier,andareonlybrieflydescribedhere.Priortoincubationwithenzymepreparations,bilberriesandblackcurrantswerethawedandmashed.A50gsampleoftheberrymashwasweighed,and5.0mLofwater–enzymesolutionwasadded.Theenzymepreparationsweredosedat1,10and100nkat/gofmash(EconaseCE,BiopectinaseCCMandPectinexBE3-L)or1,10and20nkat/gofmash(PectinexSmashXXL)basedontheirpolygalacturonase(PG)activityrepresentingenzymepreparationvolumesfrom0.03to92mL/kgberrymash.Aftertheprocessingjuiceyieldwasdeterminedbyweighingthepressedjuice,whichwascomparedtotheinitialsampleweight.Thetreatmentswerecarriedoutinfourreplicates.Controltreatmentwascarriedoutcorrespondinglybutomittingtheenzymes.Thesampleswerefrozenandstoredat–20℃untilanthocyaninanalysis.MaterialsandmethodsAnalysisofanthocyaninsFrozenjuiceswerethawed,anda4.7mLsamplewasweighedfortheanalysis.AfteradjustingthepHbelow1byadding50μLofHCl(c(HCl)=12mol/L),thesamplevolumewasadjustedto5mLwithmethanol.PriortoHPLCanalysis,thesampleswerefilteredthrougha0.45μmGHpolypropylenesyringefilter.Fortheanthocyaninspresentinunprocessedberriesthedetailsoftheextractionprocedurehavebeendescribedelsewhere.Theanalysisofanthocyaninswasperformedusing8.5%(v/v)aqueousformicacidaseluentAandHPLCgradeacetonitrile:methanol(85:15,v/v)aseluentB.Theflowratesofthemobilephasewere0.85mL/minfor0–12minand0.7mL/minfor13–90min.Thegradientusedwasasfollows:0–2min,4–6%B;2–4min,6–8%B;4–13min,8–9%B;13–20min,9–10%B;20–40min,10–11%B;40–53min,11%B;53–65min,11–19%B;65–81min19–35%B;81–84min,35–80%B,followedbyanisocraticelutionfor4minandthenreturningtotheinitialconditionsfor6minbeforethenextinjection.Anthocyaninsweredetectedat520nm.ResultsanddiscussionActivityprofileofenzymepreparationsResultsanddiscussionEffectofenzymaticprocessingonconcentrationofanthocyaninsResultsanddiscussionEffectofenzymaticprocessingonyieldofanthocyaninsResultsanddiscussionConclusion总而言之，覆盆子与黑加仑果汁进过果胶酶制剂的处理后花青素的提取率都有增加。对覆盆子而言，花青素的提取率在较高酶剂量的情况下有所提升，但100nkat/g剂量的样品相比较于10nkat/g剂量的样品而言，花青素的提取率并没有明显的增加。对黑加仑而言，花青素的最大提取率在最高的酶剂量处获得，这表明黑加仑最佳的果胶酶处理剂量在100nkat/g附近出现。因此，果胶酶的添加剂量至少应该在10nkat/g以上。谢谢！