TheRaffinose/D-Galactosetestkitisspecificandarapid measurementandanalysisofraffinoseandD-galactoseinplantmaterialsandfoodproducts.Measurementoftotalstarchincerealproductsbyamyloglucosidase-alpha-amylasemethod:collaborativestudy.McCleary,B.V.,Gibson,T.S.&Mugford,D.C.(1997).JournalofAOACInternational,80,571-579.LinktoArticleReadAbstractAnAmericanAssociationofCerealChemists/AOACcollaborativestudywasconductedtoevaluatetheaccuracyandreliABIlityofanenzymeassaykitprocedureformeasurementoftotalstarchinarangeofcerealgrainsandproducts.Thefloursampleisincubatedat95degreesCwithThermostablealpha-amylasetocatalyzethehydrolysisofstarchtomaltodextrins,thepHoftheslurryisadjusted,andtheslurryistreatedwithahighlypurifiedamyloglucosidasetoquantitativelyhydrolyzethedextrinstoglucose.Glucoseismeasuredwithglucoseoxidase-peroxidasereagent.Thirty-twocollaboratorsweresent16homogeneoustestsamplesas8blindduplicates.Thesesamplesincludedchickenfeedpellets,whitebread,greenpeas,high-amylosemaizestarch,whitewheatflour,wheatstarch,oatbran,andspaghetti.Allsampleswereanalyzedbythestandardprocedureasdetailedabove;4samples(high-amylosemaizestarchandwheatstarch)werealsoanalyzedbyamethodthatrequiresthesamplestobecookedfirstindimethylsulfoxide(DMSO).Relativestandarddeviationsforrepeatability(RSD(r))rangedfrom2.1to3.9%,andrelativestandarddeviationsforreproducibility(RSD(R))rangedfrom2.9to5.7%.TheRSD(R)valueforhighamylosemaizestarchanalyzedbythestandard(non-DMSO)procedurewas5.7%;thevaluewasreducedto2.9%whentheDMSOprocedurewasused,andthedeterminedstarchvaluesincreasedfrom86.9to97.2%.Measurementofcarbohydratesingrain,feedandfood.McCleary,B.V.,Charnock,S.J.,Rossiter,P.C.,O’Shea,M.F.,Power,A.M.&Lloyd,R.M.(2006).JournaloftheScienceofFoodandAgriculture,86(11),1648-1661.LinktoArticleReadAbstractProceduresforthemeasurementofstarch,starchdamage(gelatinisedstarch),resistantstarchandtheamylose/amylopectincontentofstarch,β-glucan,fructan,glucomannanandgalactosyl-sucroseoligosaccharides(raffinose,stachyoseandverbascose)inplantmaterial,animalfeedsandfoodsaredescribed.Mostofthesemethodshavebeensuccessfullysubjectedtointerlaboratoryevaluation.AllmethodsarebasedontheuseofenzymeseitherpurifiedbyconventionalchromatographyorproducedusingmolecularBIOLOGytechniques.Suchmethodsallowspecific,accurateandreliablequantificationofaparticularcomponent.Problemsincalculatingtheactualweightofgalactosyl-sucroseoligosaccharidesintestsamplesarediscussedindetail.Acidicα-galactosidaseisthemostabundantnectarininfloralnectarofcommontobacco(Nicotianatabacum).Zha,H.G.,Flowers,V.L.,Yang,M.,Chen,L.Y.&Sun,H.(2012).Annalsofbotany,109(4),735-745.LinktoArticleReadAbstractBackgroundandAims:Todate,mostfloralnectarins(nectarproteins)arereportedtofunctioninnectardefence,particularlyforinsect-pollinatedoutcrossingspecies.Wecomparednectarincompositionandabundanceinselfingcommontobacco(Nicotianatobaccum)withoutcrossingornamentaltobaccoplantstoelucidatethefunctionaldifferenceofnectarinsindifferentreproductivesystems.Methods:Commontobacco(CT)nectarinswereseparatedbySDS-PAGEandtheNterminusofthemostabundantnectarinwassequencedviaEdmandegradation.Thefull-lengthnectaringenewasamplifiedandclonedfromgenomicDNAandmRNAwithhiTail-PCRandRACE(rapidamplificationofCDNAends),andexpressionpatternsweretheninvestigatedindifferenttissuesusingsemi-quantitativereversetranscriptasePCR.Additionally,high-performanceliquidchromatographyandenzymaticanalysesofnectarsugarcomposition,andotherbiochemicaltraitsandfunctionsofthenovelnectarinwerestudied.KeyResults:ThemostabundantnectarininCTnectarisanacidicα-galactosidase,heredesignatedNTα-Gal.Thiscompoundhasamolecularmassof40013DaandatheoreticalpIof5•33.NTα-Galhasaconservedα-Galcharacteristicsignature,encodesamatureproteinof364aminoacidsandisexpressedindifferentorgans.Comparedwith27othermelliferousplantspeciesfromdifferentfamilies,CTfloralnectardemonstratedthehighestα-Galactivity,whichisinhibitedbyD-galactose.RaffinosefamilyoligosaccharideswerenotdetectedinCTnectar,indicatingthatNTα-Galdoesnotfunctioninpost-secretoryhydrolysis.Moreover,tobaccoplantfruitsdidnotdevelopintactskinwithgalactoseinhibitionofNTα-Galactivityinnectar,suggestingthatNTα-Galinducescell-wallsurfacerestructuringduringtheinitialstagesoffruitdevelopment.Conclusions:α-GalwasthemostabundantnectarininselfingCTplants,butwasnotdetectedinthenectarofstrictlyoutcrossingsistertobaccospecies.Nofunctionwasdemonstratedinantimicrobialdefence.Therefore,floralnectarinsinselfingspeciesmaintaintheirfunctionalsignificanceinreproductiveorgandevelopment.ViscozymeLactiononsoyslurryaffectscarbohydratesandantioxidantpropertiesofsilkentofu.Rosset,M.,Prudencio,S.H.&Beléia,A.D.P.(2012).FoodScienceandTechnologyInternational,18(6),531-538.LinktoArticleReadAbstractThisstudyinvestigatedtheenzymatictreatmentofsoyslurryusingViscozymeLtohydrolyzethecarbohydrates.TheoptimumtemperatureofViscozymeLactionwas55°C.Theincreaseofglucoseandgalactosecontentintofu(1.36and0.19 g/100 g,respectively)confirmedtheViscozymeactivityonsoyslurrywhencomparedtothecontrol.Thetreatedtofuhadmoretotalphenolicsthanthecontrol(173and161 mggallicacidequivalents/100 gfreeze-driedtofu,respectively)andhigherantioxidantactivitybythe2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonicacid)diammoniumsaltand1,1-diphenyl-2-picryhydrazyl,2,2-diphenyl-1-picryhydrazylrADIcaltests.Totalreducingsugar(glucoseequivalents)contentintreatedtofuwasapproximatelyfourtimeshigherthanthatinthecontrolundertheoptimumconditions(30FungalBeta-Glucanaseunits/10 gsolids,55°C,30 min).Thetofusdifferedinthesensoryanalysisforsoyodorandsurfaceuniformity,buttherewasnopreferenceforoneovertheother.EffectsofabrownbeanseveningmealonmetabolicriskMarkersandappetiteregulatinghormonesatasubsequentstandardizedbreakfast:arandomizedcross-overstudy.Nilsson,A.,Johansson,E.,Ekström,L.&Björck,I.(2013).PloSone,8(4),e59985.LinktoArticleReadAbstractBackground:Dietarypreventionstrategiesareincreasinglyrecognizedasessentialtocombatthecurrentepidemicofobesityandrelatedmetabolicdisorders.ThepurposeofthepresentstudywastoevaluatethepotentialprebioticeffectsofindigestIBLecarbohydratesinSwedishbrownbeans(Phaseolusvulgarisvar.nanus)inrelationtocardiometabolicriskmarkersandappetiteregulatinghormones.Methods:Brownbeans,orwhitewheatbread(WWB,referenceproduct)wereprovidedaseveningmealsto16healthyyoungadultsinarandomisedcrossoverdesign.Glucose,insulin,appetiteregulatoryhormones,GLP-1,GLP-2,appetitesensations,andmarkersofinflammationweremeasuredatafollowingstandardisedbreakfast,thatisat11to14hposttheeveningmeals.Additionally,colonicfermentationactivitywasestimatedfrommeasurementofplasmashortchainfattyacids(SCFA,includingalsobranchedchainfattyacids)andbreathhydrogen(H2)excretion.Results:Aneveningmealofbrownbeans,incomparisonwithWWB,loweredbloodglucose(−15%,ppppp=0.05),increasedGLP-2concentrations(8.4%,pp2(141%,pppConclusions:Aneveningmealwithbrownbeansbeneficiallyaffectedimportantmeasuresofcardiometabolicriskandappetiteregulatoryhormones,withinatimeframeof11–14h,incomparisontoaWWBeveningmeal.ConcentrationsofplasmaSCFAandH2wereincreased,indicatinginvolvementofcolonicfermentation.Indigestiblecolonicsubstratesfrombrownbeansmayprovideapreventivetoolinrelationtoobesityandthemetabolicsyndrome.Theeffectsoffermentationandenzymatictreatmentofpeaonnutrientdigestibilityandgrowthperformanceofbroilers.Boroojeni,F.G.,Senz,M.,Kozłowski,K.,Boros,D.,Wisniewska,M.,Rose,D.,Männer,K.&Zentek,J.(2017).Animal,1-10.LinktoArticleReadAbstractThepresentstudyexaminedtheimpactsofnative,fermentedorenzymaticallytreatedpeas(PisumsativumL.)inclusioninbroilerdiets,ongrowthperformanceandnutrientdigestibility.Forthefermentationprocess,Madonnapeawasmixedwithwater(1/1)containing2.57×108Bacillussubtilis(GalliPro®)spores/kgpeaandthen,incubatedfor48hat30°C.Fortheenzymatictreatmentprocess,theusedwaterfordoughproductioncontainedthreeenzymes,AlphaGalTM (α-galactosidase),RONOZYME®ProActandVP(proteaseandpectinasesrespectively–DSM,Switzerland)andthepeadoughincubatedfor24hat30°C.Ninecorn-wheat-soybeandietswereformulatedbysupplying10%,20%and30%oftherequiredCPwitheithernative,fermentedorenzymaticallytreatedpeas.Performancewasrecordedweeklyandattheendoftheexperiment(day35),apparentilealdigestibility(AID)ofCP,aminoacids(AA),crudefat,starch,Ca,PandKweredetermined.DataweresubjectedtoANOVAusingGLMprocedurewitha3×3factorialarrangementoftreatments.Bothprocessesreducedα-galactosides,phytate,trypsininhibitoractivityandresistantstarchinpeas.Increasinglevelsofpeaproductsupto300g/kgdiet,reducedBWgainandfeedintake(P≤0.05).Broilersfeddietscontainingenzymaticallytreatedpeahadthebestfeedconversionratioatday35.DifferenttypesofpeaproductandtheirinclusionlevelshadnoeffectonAIDofallnutrients.TheinteractionbetweentypeofthepeaproductsandinclusionlevelswassignificantforAIDofstarch.Fornativepeadiets,10%groupshowedsimilarAIDofstarchto20%nativepeabutithadhigherAIDthan30%nativepea.Forfermentedandenzymaticallytreatedgroups,allthreelevelsdisplayedsimilarAIDofstarch.Inconclusion,enzymatictreatmentandfermentationcouldimprovethenutritionalqualityofpea.Inclusionofenzymaticallytreatedpeainbroilerdietscouldimprovebroilerperformancecomparedwithotherpeaproductswhile,itdisplayedneitherpositivenornegativeimpactonnutrientdigestibility.Thepresentfindingsindicatethefeasibilityoftheseprocesses,particularlyenzymatictreatment,forimprovingthenutritionalqualityofpeaasaproteinsourceforbroilernutrition.UV-methodforthedeterminationofRaffinose(alsostachyoseandverbascose)andD-Galactoseinlegumeseeds,plantmaterials,foodstuffsandfeedPrinciple:                                 (α-galactosidase)(1)Raffinose+stachyose+verbascose+H2O→D-galactose+                                        sucrose        (galactosemutarotase)(2)α-D-Galactose↔β-D-galactose           (β-galactosedehydrogenase)(3)β-D-Galactose+NAD+→D-galactonicacid+NADH+H+Kitsize:                        *120assays* Thenumberofmanualtestsperkitcanbedoubledifallvolumesarehalved. ThiscanbereadilyaccommodatedusingtheMegaQuantTM WaveSpectrophotometer(D-MQWAVE).Method:                         Spectrophotometricat340nmReactiontime:                 ~60minDetectionlimit:                21mg/LApplicationexamples:Cerealflours,soybeanflour,by-productsofsucrosemanufactureandothermaterialsMethodrecognition:   UsedandacceptedinfoodanalysisAdvantagesVeryrapidreactionduetoinclusionofgalactosemutarotase(patentedtechnology) Verycompetitiveprice(costpertest) Allreagentsstablefor>2yearsafterpreparation Mega-Calc™softwaretoolisavailablefromourwebsiteforhassle-freerawdataprocessing Standardincluded

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