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商品描述
HighpurityArABInogalactan(LarchWood)foruseinresearch,biochemicalenzymeassaysandinvitrodiagnosticanalysis.
Purity>95%.Ara:Gal=15:85.Foruseasadietaryfiberstandard.
CompletegenomeofanewFirmicutesspeciesbelongingtothedominanthumancolonicmicrobiota(‘Ruminococcusbicirculans’)revealstwochromosomesandaselectivecapacitytoutilizeplantglucans.
Wegmann,U.,Louis,P.,Goesmann,A.,Henrissat,B.,Duncan,S.H.&Flint,H.J.(2014).EnvironmentalMicroBIOLOGy,16(9),2879–2890.
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Therecentlyisolatedbacterialstrain80/3representsoneofthemostabundant16SrRNAphylotypesdetectedinthehealthyhumanlargeintestineandbelongstotheRuminococcaceaefamilyofFirmicutes.Thecompletedgenomesequencereportedhereisthefirstforamemberofthisimportantfamilyofbacteriafromthehumancolon.Thegenomecomprisestwolargechromosomesof2.24and0.73Mbp,leADIngustoproposethenameRuminococcusbicirculansforthisnewspecies.Analysisofthecarbohydrateactiveenzymecomplementsuggestsanabilitytoutilizecertainhemicelluloses,especiallyβ-glucansandxyloglucan,forgrowththatwasconfirmedexperimentally.Theenzymaticmachineryenablingthedegradationofcelluloseandxylanbyrelatedcellulolyticruminococciishoweverlackinginthisspecies.Whilethegenomeindicatedthecapacitytosynthesizepurines,pyrimidinesandall20aminoacids,onlygenesforthesynthesisofnicotinate,NAD+,NADP+andcoenzymeAweredetectedamongtheessentialvitaminsandco-factors,resultinginmultiplegrowthrequirements.Invivo,thesegrowthfactorsmustbesuppliedfromthediet,hostorothergutmicroorganisms.OtherfeaturesofecologicalinterestincludetwotypeIVpilins,multipleextracytoplasmicfunction-sigmafactors,aureaseandabilesalthydrolase.
ProteomicinsightsintomannandegradationandproteinsecretionbytheforestfloorbacteriumChitinophagapinensis.
Larsbrink,J.,Tuveng,T.R.,Pope,P.B.,Bulone,V.,Eijsink,V.G.,Brumer,H.&McKee,L.S.(2017).JournalofProteomics,156,63-74.
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Togetherwithfungi,saprophyticbacteriaarecentraltothedecompositionandrecyclingofbiomassinforestenvironments.TheBacteroidetesphylumisabundantindiversehabitats,andseveralspecieshavebeenshowntobeabletodeconstructawidevarietyofcomplexcarbohydrates.ThegenusChitinophagaisoftenenrichedinhotspotsofplantandmicrobialbiomassdegradation.WepresentaproteomicassessmentoftheabilityofChitinophagapinensistogrowonanddegrademannanpolysaccharides,usinganagaroseplate-basedmethodofproteincollectiontominimisecontaminationwithexopolysaccharidesandproteinsfromlysedcells,andtoreflecttherealisticsettingofgrowthonasolidsurface.WeshowthatselectPolysaccharideUtilisationLoci(PULs)areexpressedindifferentgrowthconditions,andidentifyenzymesthatmaybeinvolvedinmannandegradation.Bycomparingproteomicandenzymaticprofiles,weshowevidencefortheinducedexpressionofenzymesandPULsincellsgrownonmannanpolysaccharidescomparedwithcellsgrownonglucose.Inaddition,weshowthatthesecretionofputativebiomass-degradingenzymesduringgrowthonglucosecomprisesasystemfornutrientscavenging,whichemploysconstitutivelyproducedenzymes.Significanceofthisstudy:Chitinophagapinensisbelongstoabacterialgenuswhichisprominentinmicrobialcommunitiesinagriculturalandforestenvironments,whereplantandfungalbiomassisintensivelydegraded.Suchdegradationishugelysignificantintherecyclingofcarboninthenaturalenvironment,andtheenzymesresponsIBLeareofbiotechnologicalrelevanceinemergingtechnologiesinvolvingthedeconstructionofplantcellwallmaterial.Thebacteriumhasacomparativelylargegenome,whichincludesmanyuncharacterisedcarbohydrate-activeenzymes.Wepresentthefirstproteomicassessmentofthebiomass-degradingmachineryofthisspecies,focusingonmannan,anabundantplantcellwallhemicellulose.Ourfindingsincludetheidentificationofseveralnovelenzymes,whicharepromisingtargetsforfuturebiochemicalcharacterisation.Inaddition,thedataindicatetheexpressionofspecificPolysaccharideUtilisationLoci,inducedinthepresenceofdifferentgrowthsubstrates.Wealsohighlighthowaconstitutivesecretionofenzymeswhichdeconstructmicrobialbiomasslikelyformspartofanutrientscavengingprocess.
ThetranscriptionfactorPDR-1isamulti-functionalregulatorandkeycomponentofpectindeconstructionandcatabolisminNeurosporacrassa.
Thieme,N.,Wu,V.W.,Dietschmann,A.,Salamov,A.A.,Wang,M.,Johnson,J.,Singan,V.R.,Grigoriev,I.V.,Glass,N.L.,Somerville,C.R.,&Benz,J.P.(2017).BiotechnologyforBiofuels,10(1),149.
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Background:Pectinisanabundantcomponentinmanyfruitandvegetablewastesandcouldthereforebeanexcellentresourceforbiorefinery,butiscurrentlyunderutilized.Fungalpectinasesalreadyplayacrucialroleforindustrialpurposes,suchasforfoodstuffprocessing.However,theregulationofpectinasegeneexpressionisstillpoorlyunderstood.Foranoptimalutilizationofplantbiomassforbiorefineryandbiofuelproduction,adetailedanalysisoftheunderlyingregulatorymechanismsiswarranted.Inthisstudy,weappliedthegeneticresourcesofthefilamentousascomycetespeciesNeurosporacrassatoscreenfortranscriptionfactorsthatplayamajorroleinpectinaseinduction.Results:Thepectindegradationregulator-1(PDR-1)wasidentifiedthroughatranscriptionfactormutantscreeninN.crassa.TheΔpdr-1mutantexhibitedaseveregrowthdefectonpectinandalltestedpectin-relatedpoly-andmonosaccharides.BiochemicalaswellastranscriptionalanalysesofWTandtheΔpdr-1mutantrevealedthatwhilePDR-1-mediatedgeneinductionwasdependentonthepresenceofL-rhamnose,italsostronglyaffectedthedegradationofthehomogalacturonanbackbone.Theexpressionoftheendo-polygalacturonasegh28-1wasgreatlyreducedintheΔpdr-1mutant,whiletheexpressionlevelsofallpectatelyasegenesincreased.Moreover,apdr-1overexpressionstraindisplayedsubstantiallyincreasedpectinaseproduction.PromoteranalysisofthePDR-1regulonallowedrefinementoftheputativePDR-1DNA-bindingmotif.Conclusions:PDR-1ishighlyconservedinfilamentousascomycetefungiandispresentinmanypathogenicandindustriallyimportantfungi.OurdatademonstratethatthefunctionofPDR-1inN.crassacombinesfeaturesoftworecentlydescribedtranscriptionfactorsinAspergillusniger(RhaR)andBotrytiscinerea(GaaR).Theresultspresentedinthisstudycontributetoabroaderunderstandingofhowpectindegradationisorchestratedinfilamentousfungiandhowitcouldbemanipulatedforoptimizedpectinaseproduction.
Anevolutionarilydistinctfamilyofpolysaccharidelyasesremovesrhamnosecappingofcomplexarabinogalactanproteins.
Munoz-Munoz,J.,Cartmell,A.,Terrapon,N.,Baslé,A.,Henrissat,B.&Gilbert,H.J.(2017).JournalofBiologicalChemistry,jbc-M117.
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Thehumangutmicrobiotautilizescomplexcarbohydratesasmajornutrients.Therequirementforefficientglycandegradingsystemsexertsamajorselectiveselectionpressureonthismicrobialcommunity.Thus,weproposethatthismicrobialecosystemrepresentsasubstantialresourcefordiscoveringnovelcarbohydrateactiveenzymes.TotestthishypothesiswescreenedthepotentialenzymaticfunctionsofhypotheticalproteinsencodedbygenesofBacteroidesthetaiotaomicronthatwereupregulatedbyarabinogalactanarabinogalactanproteinsorAGPs.AlthoughAGPsareubiquitousinplants,thereisapaucityofinformationontheirdetailedstructure,thefunctionoftheseglycansinplantaandthemechanismsbywhichtheyaredepolymerizedinmicrobialecosystems.HerewehavediscoveredanewpolysaccharidelyasefamilythatisspecificfortheL-rhamnose-alpha1,4-D-glucuronicacidlinkagethatcapsthesidechainsofcomplexAGPs.Thereactionproductgeneratedbythelyase,delta4,5-unsaturateduronicacid,isremovedfromAGPbyaglycosidehydrolaselocatedinfamilyGH105,producingthefinalproduct4-deoxy-β-L-threo-hex-4-enepyranosyl-uronicacid.Thecrystalstructureofamemberofthenovellyasefamilyrevealedacatalyticdomainthatdisplaysan(alpha/alpha6)6barrelfold.Inthecentreofthebarrelisadeeppocket,which,basedonmutagenesisdataandaminoacidconservation,comprisestheactivesiteofthelyase.Atyrosineistheproposedcatalyticbaseinthebeta-eliminationreaction.Thisstudyillustrateshowhighlycomplexglycanscanbeusedasascaffoldtodiscovernewenzymefamilieswithinmicrobialecosystemswherecarbohydratemetabolismisamajorevolutionarydriver.
Improvedstarchrecoveryfrompotatoesbyenzymesandreducedwaterholdingoftheresidualfibres.
Ramasamy,U.R.,Lips,S.,Bakker,R.,Gruppen,H.&Kabel,M.A.(2014).CarbohydratePolymers,113,256-263.
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Duringtheindustrialextractionofstarchfrompotatoes(Seresta),somestarchremainswithinundisruptedpotatocellsinthefibrousside-stream.Theaimofthisstudywastoinvestigateifenzymaticdegradationofcellwallpolysaccharides(CWPs)canenhancestarchrecoveryandlowerthewaterholdingcapacity(WHC)ofthe“fibre”fraction.Theuseofapectinase-richpreparationrecovered58%ofthestarchpresentinthe“fibre”fraction.Also,the“fibre”fractionretainedonly40%ofthewaterpresentinthenon-enzymetreated“fibre”.Thiswascausedbythedegradationofpectins,inparticulararabinogalactansidechainscalculatedasthesumofgalactosylandarabinosylresidues.
ComparisonofstructureandantioxidantactivityofpolysaccharidesextractedfromtheleavesofPlantagomajorL.,P.mediaL.andP.lanceolate.L.
Lukova,P.K.,Karcheva-Bahchevanska,D.P.,Nikolova,M.M.,Iliev,I.N.&Mladenov,R.D.(2017).BulgarianChemicalCommunications,49,282-288.
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InthecurrentstudyforthefirsttimewereinvestigatedthechemicalcompositionandantioxidantactivityofpolysaccharidesisolatedfromthreeindigenousforBulgariaspeciesofPlantagogenus-PlantagomajorL.,PlantagolanceolataL.andPlantagomediaL.Crudepolysaccharideswereextractedfromfreshleaveswithwateranddiluteacidandtheiryieldwasbetween0.64%and2.79%.Thechemicalcompositionofwater-extractablepolysaccharides(WEPs)andtotalacid-extractablepolysaccharides(TAEPs)ofPlantagoleaveswasevaluatedbyHPLCanalysis.Thephytochemicaldatarevealedthepresenceofbranchedheteropolysaccharideswithdifferentneutral/acidicmonosaccharideratio.ThepredominantmonosaccharideunitofWEPswasgalacturonicacid(62.64%-70.58%).Additionally,therewereregisteredsmallamountsofarabinoseandrhamnose.InTAEPsamongwithgalacturonicacid(36.93%-41.46%),significantamountsofneutralmonosaccharidesasgalactose(22.80%-46.11%)andrhamnose(16.96%-35.74%)weredetermined.TwotypesofanalyseswereusedtoevaluatetheantioxidantactivityofPlantagoisolatedpolysaccharides:DPPHandFRAPassay.BasedonDPPHmethod,WEPsexhibitedstrongerradicalscavengingability(29.39%-40.08%)comparedtoTAEPs(19.44%-24.15%).Inparallel,WEPsshowedgreaterrateofferricreducingpower(103.71-137.83µMTE/5mgPs)comparedtoTAEPs(34.63-117.66µMTE/5mgPs).AlthoughlowerthansyntheticBHT,Plantagopolysaccharidesrevealedantioxidantpotentialandcouldbefurtherexploredaspromisingnaturalantioxidantsforthenutraceuticalandpharmaceuticalindustries.
