产品资料

Megazyme/L-Glutamic Acid Assay Kit/K-GLUT/60 assays (manual) / 600 assays (microplate) / 700 assays (auto-analyser)

TheL-GlutamicAcidtestkit isasimple,reliable,rapidandaccuratemethodforthemeasurementandanalysisofL-glutamate(MSG)infoodstuffs.

Suitableformanual,auto-analyserandmicroplateformats.

SurfactanttodyebindingdegreebasedapproachfortheselectivedeterminationofL-glutamateinfoodstuffs.

Pedraza,A.,Sicilia,M.D.,Rubio,S.&Pérez-Bendito,D.(2007).AnalyticalandBioanalyticalChemistry,389(7-8),2297-2302.

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AselectivemethodforthedeterminationofL-glutamateinfoodstuffshasbeendeveloped.ItwasbasedonthecompetitionestablishedbetweentheanalyteandthedyeCoomassiebrilliantblueG(CBBG)tointeractwiththesurfactantdidodecyldimethylammoniumbromide(DDABr).ThemeasurementparameterwastheamountofDDABrrequiredtoreachagivendye-to-surfactantbindingdegree.ItwasobtainedbyphotometrictitrationonthebasisofthechangesobservedinthespectralcharacteristicsofthedyewhenCBBG–DDABraggregateswereformed.ThecalibrationgraphobtainedwaslinearintheL-glutamateconcentrationinterval0.2–5mM(detectionlimit0.05mM).Thehighselectivityoftheproposedmethod(otheraminoacidsandfoodadditivesdidnotinterfereattheconcentrationspresentinfoodstuffs)permittedthedirectanalysisoffoodsamplesafterdissolutionoftheanalyteinhotwater.TheaccuracyofthesurfactanttothedyebindingdegreemethodwasdemonstratedbydeterminingL-glutamateindifferentkindsoffoodstuffs(liquidanddriedsoups,seasonings,pastasaucesanddriedmushroomcreams)andcomparingtheresultsobtainedwiththoseprovidedbythecommercialBoehringerMannheimessay.

ThericeR2R3-MYBtranscriptionfactorOsMYB55isinvolvedinthetolerancetohightemperatureandmodulatesaminoacidmetabolism.

El-kereamy,A.,Bi,Y.M.,Ranathunge,K.,Beatty,P.H.,Good,A.G.&Rothstein,S.J.(2012).PloSone,7(12),e52030.

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Temperatureshigherthantheoptimumnegativelyaffectsplantgrowthanddevelopment.Tolerancetohightemperatureisacomplexprocessthatinvolvesseveralpathways.Understandingthisprocess,especiallyincropssuchasrice,isessentialtoprepareforpredictedclimatechangesduetoglobalwarming.Here,weshowthatOsMYB55isinducedbyhightemperatureandoverexpressionofOsMYB55resultedinimprovedplantgrowthunderhightemperatureanddecreasedthenegativeeffectofhightemperatureongrainyield.Transcriptomeanalysisrevealedanincreaseinexpressionofseveralgenesinvolvedinaminoacidsmetabolism.WedemonstratethatOsMYB55bindstothepromoterregionsoftargetgenesanddirectlyactivatesexpressionofsomeofthosegenesincludingglutaminesynthetase(OsGS1;2)glutamineamidotransferase(GAT1)andglutamatedecarboxylase3(GAD3).OsMYB55overexpressionresultedinanincreaseintotalaminoacidcontentandoftheindividualaminoacidsproducedbytheactivationoftheabovementionedgenesandknownfortheirrolesinstresstolerance,namelyL-glutamicacid,GABAandarginineespeciallyunderhightemperaturecondition.Inconclusion,overexpressionofOsMYB55improvesriceplanttolerancetohightemperature,andthishightoleranceisassociatedwithenhancedaminoacidmetabolismthroughtranscriptionactivation.

Astrocyticglutamatetransporter-dependentneuroprotectionagainstglutamatetoxicity:Aninvitrostudyofmaslinicacid.

Qian,Y.,Guan,T.,Tang,X.,Huang,L.,Huang,M.,Li,Y.,Sun,H.,Yu,R.&Zhang,F.(2011).EuropeanJournalofPharmacology,651(1-3),59-65.

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TheastrocyticglutamatetransportersGLAST/EAAT1andGLT-1/EAAT2arecrucialfortheremovalofglutamatefromthesynapticcleftandareessentialformaintainingalowconcentrationofextracellularglutamateinthebrain.Enhancedtransporterexpressionisneuroprotective.Inthepresentstudy,wetestedtheneuropotectiveeffectsofmaslinicacid,anaturalproductfromtheOleaeuropaeaplant,onculturesofprimaryneuronsfromthecerebralcortex.Studiesshowedthatastrocyte-conditionedmediumfrommaslinicacid-treatedastrocytesdose-dependentlypromotedneuronsurvivalduringglutamatetoxicitybyenhancingextracellularglutamateclearance.Real-timePCRandwesternblotanalysisrevealedthatmaslinicacidpre-treatmentsignificantlyincreasedtheexpressionofGLASTandGLT-1attheproteinandmRNAlevels.Inaddition,thisneuroprotectionwasabolishedbytheglutamatetransporterinhibitor,L-Threohydroxyaspartate(THA),inaco-cultureofastrocytesandneurons.Thesefindingssuggestthatmaslinicacidregulatestheextracellularglutamateconcentrationbyincreasingtheexpressionofastrocyticglutamatetransporters,whichmay,inturn,provideneuroprotection.

Productionofstablequininenanodispersionsusingesterifiedγ-polyglutamicacidbiopolymer.

Hoennscheidt,C.,Kreyenschulte,D.,Margaritis,A.&Krull,R.(2013).BiochemicalEngineeringJournal,79,259-266.

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NovelmethodsareneededforthedevelopmentofnanodisperseddrugformulationstoenhancebioavailABIlityofmanyhydrophobicpharmaceuticals.Thepoorlywater-solublequinineisawell-knownanti-malariadrugwhichcanbeusedasapromisingmodelcompoundforthedevelopmentofnovelnanodispersedformulations.Inadditiontohydrophobicdrug"sownaffectingproperties,surfactantsplayanimportantrolefortheenhancementoftheirlowbioavailabilitybypreparingstabledispersions.Amphiphiliccompoundscanefficientlybeusedtostabilizecolloidalfragmentsbypreventingtheprecipitationorcrystallizationofpoorlywater-solubleactiveingredientsduringfabrication.Anovelbiopolymerderivativebasedonthebiotechnologicallyproducedγ-polyglutamicacid(γ-PGA)fromBacilluslicheniformiscultivationwasdevelopedforencapsulationoftheactiveingredient.High-molecularγ-PGAisananionicpolyelectrolytethatwasoptimizedandmodifiedwithhydrophobicL-phenylalanineethylester(L-PAE)toformanamphiphiliccombpolymerP(γ-GA-r-L-PAE)withsurfactiveproperties.Theapproachofthenanodispersionpolymerconcentration,molecularweightandgraftingdegreeenablestheefficientstabilizationofthepoorlywater-solublemodeldrug.Theresearchpresentedinthisreportindicatesthepotentialbenefitsofhydrophobicallymodifiedγ-PGAandsuggestsitspotentialroleinformingstabledispersionsforfuturepharmaceuticalapplications.

Gamma-aminobutyricacid,glutamatedehydrogenaseandglutamatedecarboxylaselevelsinphylogeneticallydivergentplants.

Seher,Y.,Filiz,O.&Melike,B.(2013).PlantSystematicsandEvolution,299(2),403-412.

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Gamma-aminobutyricacid(GABA)isanonproteinaminoacidfoundinawiderangeoforganismsincludingplants.SeveralstudieshaveshownthatGABAplaysdifferentrolesinplantmetabolismincludingcarbon–nitrogenmetabolism,energybalance,signalinganddevelopment.IthasbeensuggestedthattheoccurrenceofGABAandtheenzymesrelatedtoGABAbiosynthesisinprokaryotesandeukaryotesmaybeimportantinevolutionanddiversification.However,studiesofGABAbiosynthesisandGABAlevelsinanevolutionarycontextarerestrictedtosequencedplantgenomes.InthisstudyweaimedtocomparetheactivitiesofGDHandGADenzymesandtotalnitrogen,andthecontentsoftotalsolubleprotein,succinate,glutamate,prolineandGABAinplantsfromdifferentphylogeneticlevelsincludingUlvalactuca,Pseudeverniafurfuracea,Nephrolepsisexaltata,Ginkgobiloba,Pinuspinea,Magnoliagrandiflora,Nymphaeaalba,Urticadioica,Portulacaoleraceae,Malvasylvestris,Rosacanina,Lavandulastoechas,Washingtoniafilifera,AvenabarbataandIriskaempferi.TheactivitiesofGADandGDHenzymesdifferedaccordingtothespeciesandwerenotalwaysparalleltoGABAlevels.ThediscrepancyinthecontentsofsuccinateandGABAbetweenhigherandprimitiveplantswasalsoprominent.Glutamatelevelswerehighwithafewexceptionsandprolinecontentswereatsimilarlowvaluesascomparedtootheraminoacids.OurresultssupportthehypothesisthattheGABAshuntplaysakeyroleincarbonandnitrogenpartitioningvialinkingaminoacidmetabolismandthetricarboxylicacidcyclewhichisessentialforhigherplantspecies.

Metabolicengineeringofthemixed-acidfermentationpathwayofEscherichiacoliforanaerobicproductionofglutamateanditaconate.

Vuoristo,K.S.,Mars,A.E.,Sangra,J.V.,Springer,J.,Eggink,G.,Sanders,J.P.&Weusthuis,R.A.(2015).AMBExpress,5(1),61.

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Itaconicacid,anunsaturatedC5-dicarboxylicacid,isabiobasedbuildingblockforthepolymerindustry.ThepurposeofthisstudywastoestablishproofofprincipleforananaerobicfermentationprocessfortheproductionofitaconicacidbymodificationofthemixedacidfermentationpathwayofE.coli.E.coliBW25113(DE3)andthephosphateacetyltransferase(pta)andlactatedehydrogenase(ldhA)deficientstrainE.coliBW25113(DE3)Δpta-ΔldhAwereusedtostudyanaerobicitaconateproductioninE.coli.Heterologousexpressionofthegeneencodingcis-aconitatedecarboxylase(cadA)fromA.terreusinE.coliBW25113(DE3)didnotresultinitaconateproductionunderanaerobicconditions,but0.08 mMofitaconatewasformedwhenthegenesencodingcitratesynthase(gltA)andaconitase(acnA)fromCoryNEBacteriumglutamicumwerealsoexpressed.ThesameamountwasproducedwhencadAwasexpressedinE.coliBW25113(DE3)Δpta-ΔldhA.Thetitreincreased8timesto0.66 mM(1.2 %Cmol)whenE.coliBW25113(DE3)Δpta-ΔldhAalsoexpressedgltAandacnA.Inaddition,thisstrainproduced8.5 mM(13 %Cmol)ofglutamate.Theuseofanitrogen-limitedgrowthmediumreducedtheaccumulationofglutamatebynearly50 %comparedtothenormalmedium,andalsoresultedinamorethan3-foldincreaseoftheitaconatetitreto2.9 mM.Theseresultsdemonstratedthat E.coli haspotentialtoproduceitaconateandglutamateunderanaerobicconditions,closingtheredoxbalancebyco-productionofsuccinateorethanolwithH₂ andCO₂.

Genetic,enzymaticandmetaboliteprofilingoftheLactobacilluscaseigrouprevealsstrainbiodiversityandpotentialapplicationsforflavourdiversification.

Stefanovic,E.,Kilcawley,K.N.,Rea,M.C.,Fitzgerald,G.F.&McAuliffe,O.(2017).JournalofAppliedMicroBIOLOGy,122(5),1245-1261.

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Aims:TheLactobacilluscaseigrouprepresentsawidelyexploredgroupoflacticacidbacteria,characterizedbyahighlevelofbiodiversity.Inthisstudy,thegeneticandphenotypicdiversityofacollectionofmorethan300isolatesoftheLact.caseigroupandtheirpotentialtoproducevolatilemetabolitesimportantforflavourdevelopmentindairyproducts,wasexamined.MethodsandResults:Followingconfirmationofspeciesby16SrRNAPCR,thediversityoftheisolateswasdeterminedbypulsed-fieldgelelectrophoresis.Theactivitiesofenzymesinvolvedintheproteolyticcascadewereassessedandsignificantdifferencesamongthestrainswereobserved.Tenstrainswerechosenbasedontheresultsoftheirenzymesactivitiesandtheywereanalysedfortheirabilitytoproducevolatilesinmediawithincreasedconcentrationsofarepresentativearomatic,branchedchainandsulphuraminoacid.Volatileswereassessedusinggaschromatographycoupledwithmassspectrometry.Strain-dependentdifferencesintherangeandtypeofvolatilesproducedwereevident.Conclusions:StrainsoftheLact.caseigrouparecharacterizedbygeneticandmetabolicdiversitywhichsupportsvariabilityinvolatileproduction.SignificanceandImpactoftheStudy:Thisstudyprovidesascreeningapproachfortheknowledge-basedselectionofstrainspotentiallyenablingflavourdiversificationinfermenteddairyproducts.

ColourimetricmethodforthedeterminationofL-GlutamicAcid
(MonosodiumGlutamate;MSG)infoodstuffsandothermaterials

Principle:
(beefliverglutamatedehydrogenase)
(1)L-Glutamicacid+NAD⁺+H₂O↔2-oxoglutarate+NADH+NH₄⁺

(diaphorase)
(2)INT+NADH+H⁺→NAD⁺+INT-formazan

Kitsize: * 60assays(manual)/600(microplate)
/700(auto-analyser)

* Thenumberofmanualtestsperkitcanbedoubledifallvolumesarehalved.
ThiscanbereADIlyaccommodatedusingtheMegaQuant^TMWave
Spectrophotometer(D-MQWAVE).

Method:                          Spectrophotometricat492nm
Reactiontime:                 ~8min
Detectionlimit:                0.21mg/L
Applicationexamples:
Fruitandvegetables(e.g.tomato),processedfruitandvegetables
(e.g.tomatopuree/juice,ketchup,soysauce),condiments,processed
meatproducts(e.g.extracts,bouillonandsausages),soup,pharmaceuticals
andothermaterials(e.g.biologicalcultures,samples,etc.)
Methodrecognition:
MethodsbasedonthisprinciplehavebeenacceptedbyISO,GOST
andNMKL

Advantages

Verycompetitiveprice(costpertest)
Allreagentsstablefor>2yearsafterpreparation
Glutamatedehydrogenasesolutionstableat-20°C
Nowasteddiaphorasesolution(stablesUSPensionsupplied)
Rapidreaction
Mega-Calc™softwaretoolisavailablefromourwebsiteforhassle-freerawdataprocessing
Standardincluded
Suitableformanual,microplateandauto-analyserformats

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Megazyme/L-Glutamic Acid Assay Kit/K-GLUT/60 assays (manual) / 600 assays (microplate) / 700 assays (auto-analyser)

SurfactanttodyebindingdegreebasedapproachfortheselectivedeterminationofL-glutamateinfoodstuffs.

ThericeR2R3-MYBtranscriptionfactorOsMYB55isinvolvedinthetolerancetohightemperatureandmodulatesaminoacidmetabolism.

Astrocyticglutamatetransporter-dependentneuroprotectionagainstglutamatetoxicity:Aninvitrostudyofmaslinicacid.

Productionofstablequininenanodispersionsusingesterifiedγ-polyglutamicacidbiopolymer.

Gamma-aminobutyricacid,glutamatedehydrogenaseandglutamatedecarboxylaselevelsinphylogeneticallydivergentplants.

Metabolicengineeringofthemixed-acidfermentationpathwayofEscherichiacoliforanaerobicproductionofglutamateanditaconate.

Genetic,enzymaticandmetaboliteprofilingoftheLactobacilluscaseigrouprevealsstrainbiodiversityandpotentialapplicationsforflavourdiversification.