TheAceticAcid(ACS ManualFormat)testkitisa simplemethodfortherapidandreliablemeasurementandanalysisofaceticacid/acetateinfoods,beveragesandothermaterials.
Grapeandwineanalysis:Oenologiststoexploitadvancedtestkits.
Charnock,S.C.&McCleary,B.V.(2005).RevuedesEnology,117,1-5.
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Itiswithoutdoubtthattestingplaysapivotalrolethroughoutthewholeofthevinificationprocess.Toproducethebestposs
IBLequalitywineandtominimiseprocessproblemssuchas“stuck”fermentationortroublesomeinfections,itisnowrecognisedthatifpossibletestingshouldbeginpriortoharvestingofthegrapesandcontinuethroughtobottling.Tr
ADItionalmethodsofwineanalysisareoftenexpensive,timeconsuming,requireeitherelaborateequipmentorspecialistexpertiseandfrequentlylackaccuracy.However,enzymaticbio-analysisenablestheaccuratemeasurementofthevastmajorityofanalytesofinteresttothewinemaker,usingjustonepieceofapparatus,thespectrophotometer(
seepreviousissueNo.116foradetailedtechnicalreview).Grapejuiceandwineareamenabletoenzymatictestingasbeingliquidstheyarehomogenous,easytomanipulate,andcangenerallybeanalysedwithoutanysamplepreparation.
Megazyme“advanced”winetestkitsgeneralcharacteristicsandvalidation.
Charnock,S.J.,McCleary,B.V.,Daverede,C.&Gallant,P.(2006).ReveuedesOenologues,120,1-5.
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ManyoftheenzymatictestkitsareofficialmethodsofprestigiousorganisationssuchastheAssociationofOfficialAnalyticalChemicals(AOAC)andtheAmericanAssociationofCerealChemists(AACC)inresponsetotheinterestfromoenologists.Megazymedecidedtouseitslonghistoryofenzymaticbio-analysistomakeasignificantcontributiontothewineindustry,bythedevelopmentofarangeofadvancedenzymatictestkits.Thistaskhasnowbeensuccessfullycompletedthroughthestrategicandcomprehensiveprocessofidentifyinglimitationsofexistingenzymaticbio-analysistestkitswheretheyoccurred,andthenusingadvancedtechniques,suchasmolecular
BIOLOGy(
photo1),torapidlyovercomethem.Noveltestkitshavealsobeendevelopedforanalytesofemerginginteresttotheoenologist,suchasyeastavailablenitrogen(
YAN;seepages2-3ofissue117article),orwherepreviouslyenzymesweresimplyeithernotavailable,orweretooexpensivetoemploy,suchasforD-mannitolanalysis.
MicrobialandphysicochemicalsuccessioninfermentedsausagesproducedwithbacteriocinogeniccultureofLactobacillussakeiandsemi-purifiedbacteriocinmesenterocinY.
Zdolec,N.,Hadžiosmanović,M.,Kozačinski,L.,Cvrtila,Ž.,Filipović,I.,Škrivanko,M.&Leskovar,K.(2008).MeatScience, 80(2),480-487.
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TheinfluenceofthebacteriocinogeniccultureLactobacillussakei(105/g)andsemi-purifiedbacteriocinmesenterocinY(2560AU/kg)onthesafetyandqualityoftraditionalCroatianfermentedsausageswasinvestigated.TheadditionofLb.sakeiand/ormesenterocinYreducedmicrobialcounts(P<0.05)=""in=""the=""final=""products.=""after=""28=""days=""of=""ripening,=""coagulase-negative="">coccidecreased1.5–2.0log,yeasts1.2–1.4logandenterococci1.7–2.7log.InthecaseoftheadditionofLb.sakei,thelacticacidbacteriacountwassignificantly(P<0.05)=""higher=""at=""day=""7=""of=""ripening,=""and=""was=""accompanied=""by=""a=""lower=""ph=""and=""a=""higher=""amount=""of=""lactic=""acid="">P<0.05).=""in=""the=""final=""product=""the=""amount=""of=""acetic=""acid=""was=""significantly=""lower.=""more=""intensive=""proteolysis=""and=""an=""increase=""in=""ammonia=""content=""were=""found=""at=""the=""beginning=""of=""fermentation,=""and=""in=""the=""second=""phase=""of=""ripening=""in=""the=""control=""samples,=""respectively.=""the=""free=""fatty=""acid=""concentration=""was=""significantly=""lower=""during=""the=""entire=""ripening=""process=""compared=""to=""the=""control="">P<0.05).=""semi-purified=""mesenterocin=""y=""did=""not=""affect=""the=""sensory=""properties=""of=""the=""sausages,=""whilst=""the=""addition=""of="">Lb.sakeienhancedthem.
Potentialofthewastefrombeerfermentationbrothforbio-ethanolproductionwithoutanyadditionalenzyme,microbialcellsandcarbohydrates.
Ha,J.H.,Shah,N.,Ul-Islam,M.&Park,J.K.(2011).EnzymeandMicrobialTechnology,49(3),298-304.
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Thepotentialofthewastefrombeerfermentationbroth(WBFB)fortheproductionofbio-ethanolusingasimultaneoussaccharificationandfermentationprocesswithoutanyextraadditionsofsaccharificationenzymes,microbialcellsorcarbohydratewastested.ThemajormicrobialcellsinWBFBwereisolatedandidentified.ThevariationsincompositionsofWBFBwithstocktimewereinvestigated.TherewasresidualactivityofstarchhydrolyzingenzymesinWBFB.Theeffectsofreactionmodes,e.g.staticandshakingonbio-ethanolproductionwerestudied.After7daysofcultivationusingthesupernatantofWBFBat30°Ctheethanolconcentrationreached103.8g/Linshakingcultureand91.5g/Linstaticculture.Agitationexperimentsconductedatatemperature-profileprocessinwhichtemperaturewasincreasedfrom25to67°Cshortenedthesimultaneousprocesstime.TheoriginalWBFBwasmoreusefulthanthesupernatantofWBFBingettingthehigherconcentrationofethanolandreducingthefermentationtime.FromthiswholestudyitwasfoundthatWBFBisacheapandsuitablesourceforbio-ethanolproduction.
DeterminationofOrtho‐andRetronasalDetectionThresholdsandOdorImpactof2,5‐Dimethyl‐3‐MethoxypyrazineinWine.
Botezatu,A.&Pickering,G.J.(2012).JournalofFoodScience,77(11),S394-S398.
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2,5-Dimethyl-3-methoxypyrazine(DMMP)hasbeenrecentlyidentifiedinbothCoccinellidae-tainted(byeitherCoccinellaseptempunctataorHarmoniaaxyridisbeetles)anduntaintedwines;however,littleisknownregardingitsimpactonwinearomaandflavor.Theaimsofthisstudyweretoobtainanaccurateestimateofboththeortho-andretronasaldetectionthresholdsofDMMPinredwineandtounderstandhowDMMPcontributestothearomaprofileofredwine.Inthefirststudy,thresholdsweredeterminedfor21individualsusingtheASTME679ascendingforcedchoicemethodoflimits.Theorthonasalgroupbestestimatethreshold(BET)was31ng/LandtheretronasalgroupBETwas70ng/L.Amoderatevariationinindividualthresholdswasobservedfortheorthonasalmodality[standarddeviation(SD)=19.8]andalargervariationwasnotedforretronasalthresholds(SD=111.8).Inthesecondstudy,apanelof8assessorsperformeddescriptivesensoryanalysison3redwinescontainingvariousconcentrationsofaddedDMMP(0,50and120ng/L).Resultsshowsignificantchangesinaromacharacteristicsinthe120ng/Lwineandsmallereffectsatthe50ng/Llevel.Overall,winesspikedwithDMMPgeneratedlowerintensityratingsforcherryandredberrydescriptorsandhigherratingsforearthy/mustyandgreen/vegetaldescriptors.WhenconsideredwithotherrecentresultsonDMMPconcentrationsfoundinwine,DMMPcanbeconsideredahithertoundescribedimpactodorantinsomewinestyles.
EffectsofyeastextractanddifferentaminoacidsonthedynamicsofsomecomponentsincabbagejuiceduringfermentationwithBifidobacteriumlactisBB-12.
Buruleanu,C.L.,Nicolescu,C.L.,Avram,D.,Manea,I.&Bratu,M.G.(2012).FoodScienceandBiotechnology,21(3),691-699.
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Variousaminoacidsandtheyeastextract,inamountsof0.1%(w/v),wereseparatelytestedfortheirinfluenceontheanalyticalparametersoflacticacidfermentationofcabbagejuicewithBifidobacteriumanimalissubsp.lactisBB-12.Comparedwiththecontrol,cysteinesupplementationledtoadecreaseofthetimetoreachpH5.0of6timesandanincreaseoflacticacidproductivityof1.22times.After48htheascorbicacidcontentwasby360.73%higher,thefermentedcabbagejuicesbeingassignedintoadistinctgroupapplyingbothfactoranalysis(FA)andclusteranalysis(CA).Tryptophancontributedtobettervaluesforlacticandaceticacidyield,whilelysineandyeastextractespeciallyforaceticacidyield.Valineandleucinewerenotabletoimprovethefermentationprogress,estimatedthroughtheanalyzedvariables.Thisworkwouldprovidesomehelpfulinformationforthedevelopmentofvariouslacto-fermentedvegetablejuicesusingprobioticbacteria.
Heterofermentativeprocessindryfermentedsausages-acasereport.
Kameník,J.,Dušková,M.,Saláková,A.&Šedo,O.(2013).ActaVeterinariaBrno,82(2),181-186.
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Incertaincircumstancesthefermentationprocessindryfermentedsausagesconvertstoheterofermentationpathwayleadingtoaceticacidandcarbondioxidebesidelacticacid.Thestudydescribestwocasesofundesirableheterofermentationindrysausagesfromtwodifferentproducers.Inthesausagesamples(n=7)thepHvalueandthecontentoflacticandaceticacidsweremeasured.Microbialanalysisfocusedonquantitativeandqualitativedetectionoflacticacidbacteria.Theaceticacidcontentvariedfrom24.28to67.41µmolg-1drymatter,inthecaseofsamplesfromthesecondproducerthecontentofaceticacid(48.45to67.41µmolg-1drymatter)washigherthanthelacticacidcontent(20.98to29.02µmolg-1drymatter).ThelactobacillistrainsfromthesausageswereassignedtothecorrespondingspeciesbyMatrix-AssistedLaserDesorption-Ionization-TimeofFlightMassSpectrometry(MALDI-TOFMS)andclassifiedtothreegroupsaccordingtothesugarfermentationpattern(obligatelyhomofermentative,facultativelyheterofermentativeandobligatelyheterofermentative)andtheycausedtheheterofermentationprocessinthesamplesofdryfermentedsausages.Thedescriptionofthecaseofheterofermentationprocessindrysausagesisuniqueandthereislittleinformationaboutthistopic.
Effectsofharvestdate,wiltingandinoculationonyieldandforagequalityofensilingsafflower(CarthamustinctoriusL.)biomass.
Cazzato,E.,Laudadio,V.,Corleto,A.&Tufarelli,V.(2011).JournaloftheScienceofFoodandAgriculture,91(12),2298-2302.
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BACKGROUND:Safflower(CarthamustinctoriusL.),usuallygrownasasourceofoilcrop,canbeusedasfoddereitherforhayorensilingpurposes,particularlyinsemi-aridregions.RESULTS:A2-yeartrialwasconductedinsouthernItalytoevaluatetheproductionandforagequalityofsafflowerbiomasscv.Centennial,harvestedatthreedifferentstages:1,atcompleteappearanceofprimarybuds(PB);2,atcompleteappearanceofsecondaryandtertiarybuds(STB);and3,at25%offloweringstage(FS).Foreachstageofgrowth,50%ofthebiomasswasensiledin4Lglassjarswithoutandwithinoculation(Lactobacillusplantarum,LAB),andtheother50%wasfieldwiltedfor24hbeforeensiling.Drymatter(DM)contentandyield(DMY),pH,bufferingcapacity(BC)andwatersolublecarbohydrates(WSC)weredeterminedonfreshforage.Onsafflowersilageswerealsoevaluatedammonia-N,crudeprotein(CP),fibrefractions,fat,lacticandaceticacids,CaandP,andgaslosses.DMYrangedfrom4.5tha-1(PBharvesting)to11.6tha-1(FSharvesting).DMcontentvariedfrom129gkg-1(PBnotwilted)to630gkg-1(FSwilted).TheWSCinforagebeforeensilingwithnotwiltingrangedfrom128(PBstage)to105and100gkg-1DMatSTBandFSstages,respectively.ThewiltedsafflowerforageshowedalowerWSCcomparedtowiltedforage.Thehighsugarsubstrateallowedlacticacidfermentationandagoodconservationqualityinalltheharvestingstages.Silagesqualitywasstronglyinfluencedbythetreatmentperformed.WiltingpracticeincreasedDM,pHandNDFcontentsbutreducedlacticacid,aceticacidandNH3-Nvalues.InoculationreducedDM,pHandNDFcontents,butincreasedlacticandaceticacids,CPandash.CONCLUSION:Asresult,wiltingtheforagefor1daywasveryeffectiveintheearlyharvestingstagebecausethispracticesignificantlyincreasedDM,reducingonthesametimetheintensivefermentationandproteolysisprocessesofsilage.Whenharvestingisperformedatthebeginningofthefloweringstagewiltingisnotnecessary.
LactosefermentationbyKombucha–aprocesstoobtainnewmilk–basedbeverages.
Iličić,M.,Kanurić,K.,Milanović,S.,Lončar,E.,Djurić,M.&Malbaša,R.(2012).RomanianBiotechnologicalLetters,17(1),7013-7021.
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Thispaperfocusesonfermentationoflactosefromamodelsystem(blacktea)andfromtwotypesofmilk(0.9%w/wand2.2%w/woffat)byapplicationofKombucha.QuantitiesoftheappliedKombuchastarterwere10%v/vand15%v/v.Allfermentationswereperformedat42°C.TheprocesstoachieveadesirablepH=4.5wasslowerinthemodelsystem(16h)thaninmilks(9-10h).Regardingstarterquantity,10%v/vprovedtheoptimal.Regardingtypesofmilk,higherfatcontentguaranteesshorterfermentationandhigheryieldofmetabolites.Utilizationoflactosewasfoundatalevelof≈20%and≈30%inmilkswith0.9%w/wand2.2%w/woffat,respectively.Thiswascorrelatedwithanappearanceofintermediatesand/orproducts.Glucoseunderwentfurthertransformationsalmostentirely,whilegalactoseshowedmuchlowerreactivity.Seventotwelvetimeshighercontentsoflacticacidwerefoundcomparedtoaceticacid.Milk-basedbeveragefromthereducedfatsample,inoculatedwith10%v/vofKombuchastarter,hasthebestphysicalcharacteristics(syneresisandwaterholdingcapacity).Italsodevelopedagoodtexture(especiallycohesivenessandindexofviscosity).Milklactosefermentationwasaprocessthatcouldhavebeenusedforobtainingnewmilk-basedproducts.
Deepsequencingofvoodoolily(Amorphophalluskonjac):anapproachtoidentifyrelevantgenesinvolvedinthesynthesisofthehemicelluloseglucomannan.
Gille,S.,Cheng,K.,Skinner,M.E.,Liepman,A.H.,Wilkerson,C.G.&Pauly,M.(2011).Planta,234(3),515-526.
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A
Roche454
CDNAdeepsequencingexperimentwasperformedonadevelopingcormof
Amorphophalluskonjac—alsoknownasvoodoolily.Thedominantstoragepolymerinthecormofthisplantisthepolysaccharideglucomannan,ahemicelluloseknowntoexistinthecellwallsofhigherplantsandamajorcomponentofplantbiomassderivedfromsoftwoods.Atotalof246megabasepairsofsequencedatawasobtainedfromwhich4,513distinctcontigswereassembled.Withinthisvoodoolilyexpressedsequencetagcollectiongenesrepresentingthecarbohydraterelatedpathwayofglucomannanbiosynthesiswereidentified,includingsucrosemetabolism,nucleotidesugarconversionpathwaysfortheformationofactivatedprecursorsaswellasaputativeglucomannansynthase.
Invivoexpressionoftheputativeglucomannansynthaseandsubsequentinvitroactivityassaysunambiguouslydemonstratethattheenzymehasindeedglucomannanmannosyl-andglucosyltransferaseactivities.BasedontheexpressedsequencetaganalysishithertounknownpathwaysforthesynthesisofGDP-glucose,anecessaryprecursorforglucomannanbiosynthesis,couldbeproposed.Moreover,theresultshighlighttranscriptionalbottlenecksforthesynthesisofthishemicellulose.
Functionalandanioniccellulose-interactingpolymersbyselectivechemo-enzymaticcarboxylationofgalactose-containingpolysaccharides.
Parikka,K.,Leppänen,A.S.,Xu,C.,Pitkänen,L.,Eronen,P.,Österberg,M.,Brumer,H.,Willför,S.&Tenkanen,M.(2012).Biomacromolecules,13(8),2418-2428.
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Carboxylated,anionicpolysaccharideswereselectivelypreparedusingacombinationofenzymaticandchemicalreactions.Thegalactose-containingpolysaccharidesstudiedweresprucegalactoglucomannan,guargalactomannan,andtamarindgalactoxyloglucan.Thegalactosylunitsofthepolysaccharideswerefirstoxidizedwithgalactoseoxidase(EC1.1.3.9)andthenselectivelycarboxylated,resultinginthegalacturonicacidderivativeswithgoodconversionandyield.Thedegreesofoxidation(DO)oftheproductsweredeterminedbygaschromatography–massspectrometry(GC-MS).Anovelfeasibleelectrosprayionization-massspectrometry(ESI-MS)methodwasalsodevelopedforthedeterminationofDO.Thesolutionpropertiesandchargedensitiesoftheproductswereinvestigated.Theinteractionoftheproductswithcellulosewasstudiedbytwomethods,bulksorptionontobleachedbirchkraftpulpandadsorptionontonanocelluloseultrathinfilmsbyquartzcrystalmicrobalancewithdissipation(QCM-D).Tostudytheeffectofthelocationofthecarboxylicacidgroupsonthephysicochemicalproperties,polysaccharideswerealsooxidizedby2,2,6,6-tetramethylpiperidine-1-oxyl(TEMPO)-mediatedreactionproducingpolyuronicacids.Thechemo-enzymaticallyoxidizedgalacturonicpolysaccharideswithanunmodifiedbackbonehadabetter
ABIlitytointeractwithcellulosethantheTEMPO-oxidizedproducts.Theselectivelycarboxylatedpolysaccharidescanbefurtherexploited,assuch,orinthetargetedfunctionalizationofcellulosesurfaces.
DistinctrolesofcarbohydrateesterasefamilyCE16acetylesterasesandpolymer-actingacetylxylanesterasesinxylandeacetylation.
Koutaniemi,S.,vanGool,M.P.,Juvonen,M.,Jokela,J.,Hinz,S.W.,Schols,H.A.&Tenkanen,M.(2013).JournalofBiotechnology,168(4),684-692.
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Massspectrometricanalysiswasusedtocomparetherolesoftwoacetylesterases(AE,carbohydrateesterasefamilyCE16)andthreeacetylxylanesterases(AXE,familiesCE1andCE5)indeacetylationofnaturalsubstrates,neutral(linear)and4-O-methylglucuronicacid(MeGlcA)substitutedxylooligosaccharides(XOS).AEsweresimilarlyrestrictedintheiractionandapparentlyremovedinmostcasesonlyoneacetylgroupfromthenon-reducingendofXOS,actingasexo-deacetylases.Incontrast,AXEscompletelydeacetylatedlongerneutralXOSbuthaddifficultieswiththeshorterones.CompletedeacetylationofneutralXOSwasobtainedafterthecombinedactionofAEsandAXEs.MeGlcAsubstituentspartiallyrestrictedtheactionofbothtypesofesterasesandtheremainingacidicXOSweremainlysubstitutedwithoneMeGlcAandoneacetylgroup,supposedlyonthesamexylopyranosylresidue.Theseresistingstructuresweredegradedtogreatextentonlyafterinclusionofα-glucuronidase,whichactedwiththeesterasesinasynergisticmanner.Whenusedtogetherwithxylanbackbonedegradingendoxylanaseandβ-xylosidase,bothAEandAXEenhancedthehydrolysisofcomplexXOSequally.
Pilot-scalecultivationofwall-deficienttransgenicChlamydomonasreinhardtiistrainsexpressingrecombinantproteinsinthechloroplast.
Zedler,J.A.,Gangl,D.,Guerra,T.,Santos,E.,Verdelho,V.V.&Robinson,C.(2016).AppliedMicrobiologyandBiotechnology,1-10.
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Microalgaehaveemergedaspotentiallypowerfulplatformsfortheproductionofrecombinantproteinsandhigh-valueproducts.Chlamydomonasreinhardtiiisapotentiallyimportanthostspeciesduetotherangeofgenetictoolsthathavebeendevelopedforthisunicellulargreenalga.Transformationofthechloroplastgenomeoffersimportantadvantagesovernucleartransformation,andawiderangeofrecombinantproteinshavenowbeenexpressedinthechloroplastsofC.reinhardtiistrains.Thisisoftendoneincellwall-deficientmutantsthatareeasiertotransform.However,onlyasinglestudyhasreportedgrowthdataforC.reinhardtiigrownatpilotscale,andthegrowthofcellwall-deficientstrainshasnotbeenreportedatall.Here,wereportthefirstpilot-scalegrowthstudyfortransgenic,cellwall-deficientC.reinhardtiistrains.StrainsexpressingacytochromeP450(CYP79A1)orbifunctionalditerpenesynthase(cis-abienolsynthase,TPS4)weregrownfor7daysundermixotrophicconditionsinaTris-acetate-phosphatemedium.Thestrainsreacheddrycellweightsof0.3g/Lwithin3–4dayswithstableexpressionlevelsoftherecombinantproteinsduringthewholeupscalingprocess.Thestrainsprovedtobegenerallyrobust,despitethecellwall-deficientphenotype,butgrewpoorlyunderphototrophicconditions.Thedataindicatethatcellwall-deficientstrainsmaybehighlyamenablefortransformationandsuitableforcommercial-scaleoperationsundermixotrophicgrowthregimes.