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HighpurityRhamnogalacturonan(SoyBean)foruseinresearch,biochemicalenzymeassaysandinvitrodiagnosticanalysis.
Preparedfromsoybeanpectin.Potentialsubstratefortheassayofrhamnogalacturonase.
Arevisedarchitectureofprimarycellwallsbasedonbiomechanicalchangesinducedbysubstrate-specificendoglucanases.
Park,Y.B.&Cosgrove,D.J.(2012).PlantPhysiology,158(4),1933-1943.
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Xyloglucaniswidelybelievedtofunctionasatetherbetweencellulosemicrofibrilsintheprimarycellwall,limitingcellenlargementbyrestrictingtheABIlityofmicrofibrilstoseparatelaterally.Totestthebiomechanicalpredictionsofthis“tetherednetwork”model,weassessedtheabilityofcucumber(Cucumissativus)hypocotylwallstoundergocreep(long-term,irreversIBLeextension)inresponsetothreefamily-12endo-β-1,4-glucanasesthatcanspecificallyhydrolyzexyloglucan,cellulose,orboth.Xyloglucan-specificendoglucanase(XEGfromAspergillusaculeatus)failedtoinducecellwallcreep,whereasanendoglucanasethathydrolyzesbothxyloglucanandcellulose(Cel12AfromHypocreajecorina)inducedahighcreeprate.Acellulose-specificendoglucanase(CEGfromAspergillusniger)didnotcausecellwallcreep,eitherbyitselforincombinationwithXEG.Testswithadditionalenzymes,includingafamily-5endoglucanase,confirmedtheconclusionthattocausecreep,endoglucanasesmustcutbothxyloglucanandcellulose.Similarresultswereobtainedwithmeasurementsofelasticandplasticcompliance.BothXEGandCel12Ahydrolyzedxyloglucaninintactwalls,butCel12AcouldhydrolyzeaminorxyloglucancompartmentrecalcitranttoXEGdigestion.XyloglucaninvolvementintheseenzymeresponseswasconfirmedbyexperimentswithArabidopsis(Arabidopsisthaliana)hypocotyls,whereCel12Ainducedcreepinwild-typebutnotinxyloglucan-deficient(xxt1/xxt2)walls.Ourresultsareincompatiblewiththecommondepictionofxyloglucanasaload-bearingtetherspanningthe20-to40-nmspacingbetweencellulosemicrofibrils,buttheydoimplicateaminorxyloglucancomponentinwallmechanics.Thestructurallyimportantxyloglucanmaybelocatedinlimitedregionsoftightcontactbetweenmicrofibrils.
Real-timeimagingofcellulosereorientationduringcellwallexpansioninArabidopsisroots.
Anderson,C.T.,Carroll,A.,Akhmetova,L.&Somerville,C.(2010).PlantPhysiology,152(2),787-796.
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Celluloseformsthemajorload-bearingnetworkoftheplantcellwall,whichsimultaneouslyprotectsthecellanddirectsitsgrowth.Althoughtheprocessofcellulosesynthesishasbeenobserved,littleisknownaboutthebehaviorofcelluloseinthewallaftersynthesis.UsingPontamineFastScarlet4B,adyethatfluorescespreferentiallyinthepresenceofcelluloseandhasexcitationandemissionwavelengthssuitableforconfocalmicroscopy,weimagedthearchitectureanddynamicsofcelluloseinthecellwallsofexpandingrootcells.WefoundthatcelluloseexistsinArabidopsis(Arabidopsisthaliana)cellwallsinlargefibrillarbundlesthatvaryinorientation.Duringanisotropicwallexpansioninwild-typeplants,weobservedthatthesecellulosebundlesrotateinatransversetolongitudinaldirection.Wealsofoundthatcelluloseorganizationissignificantlyalteredinmutantslackingeitheracellulosesynthasesubunitortwoxyloglucanxylosyltransferaseisoforms.Ourresultssupportamodelinwhichcelluloseisdepositedtransverselytoaccommodatelongitudinalcellexpansionandreorientedduringexpansiontogenerateacellwallthatisfortifiedagainststrainfromanydirection.
X4modulesrepresentanewfamilyofcarbohydrate-bindingmodulesthatdisplaynovelproperties.
Bolam,D.N.,Xie,H.,Pell,G.,Hogg,D.,Galbraith,G.,Henrissat,B.&Gilbert,H.J.(2004).JournalofBIOLOGicalChemistry,279(22),22953-22963.
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Thehydrolysisoftheplantcellwallbymicrobialglycosidehydrolasesandesterasesistheprimarymechanismbywhichstoredorganiccarbonisutilizedinthebiosphere,andthustheseenzymesareofconsiderablebiologicalandindustrialimportance.Plantcellwall-degrADIngenzymesingeneraldisplayamodulararchitecturecomprisingcatalyticandnon-catalyticmodules.TheX4modulesinglycosidehydrolasesrepresentalargefamilyofnon-catalyticmoduleswhosefunctionisunknown.HereweshowthattheX4modulesfromaCellvibriojaponicusmannanase(Man5C)andarabinofuranosidase(Abf62A)bindtopolysaccharides,andthustheseproteinscompriseanewfamilyofcarbohydrate-bindingmodules(CBMs),designatedCBM35.TheMan5C-CBM35bindstogalactomannan,insolubleamorphousmannan,glucomannan,andmanno-oligosaccharidesbutdoesnotinteractwithcrystallinemannan,cellulose,cello-oligosaccharides,orotherpolysaccharidesderivedfromtheplantcellwall.Man5C-CBM35alsopotentiatesmannanaseactivityagainstinsolubleamorphousmannan.Abf62A-CBM35interactswithunsubstitutedoat-speltxylanbutnotsubstitutedformsofthehemicelluloseorxylo-oligosaccharides,andrequirescalciumforbinding.Thisisinsharpcontrasttootherxylan-bindingCBMs,whichinteractinacalcium-independentmannerwithbothxylo-oligosaccharidesanddecoratedxylans.
Family42carbohydrate-bindingmodulesdisplaymultiplearabinoxylan-bindinginterfacespresentingdifferentligandaffinities.
Ribeiro,T.,Santos-Silva,T.,Alves,V.D.,Dias,F.M.V.,Luís,A.S.,Prates,J.A.M.,Ferraira,L.M.A.,Romao,M.J.&Fontes,C.M.G.A.(2010).BiochimicaetBiophysicaActa(BBA)-ProteinsandProteomics,1804(10),2054-2062.
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Enzymesthatdegradeplantcellwallpolysaccharidesdisplayamodulararchitecturecomprisingacatalyticdomainboundtooneormorenon-catalyticcarbohydrate-bindingmodules(CBMs).CBMsdisplayconsiderablevariationinprimarystructureandaregroupedinto59sequence-basedfamiliesorganizedintheCarbohydrate-ActiveenZYme(CAZy)database.HerewereportthecrystalstructureofCtCBM42Atogetherwiththebiochemicalcharacterizationoftwoothermembersoffamily42CBMsfromClostridiumThermocellum.CtCBM42A,CtCBM42BandCtCBM42Cbindspecificallytothearabinoseside-chainsofarabinoxylansandarabinan,suggestingthatvariouscellulosomalcomponentsaretargetedtotheseregionsoftheplantcellwall.ThestructureofCtCBM42Adisplaysabeta-trefoilfold,whichcomprises3sub-domainsdesignatedasα,βandγ.Eachoneofthethreesub-domainspresentsaputativecarbohydrate-bindingpocketwhereanaspartateresiduelocatedinacentralpositiondominatesligandrecognition.Intriguingly,theγsub-domainofCtCBM42Aispivotalforarabinoxylanbinding,whiletheconcertedactionofβandγsub-domainsofCtCBM42BandCtCBM42Cisapparentlyrequiredforligandsequestration.Thus,thisworkrevealsthatthebindingmechanismofCBM42membersisincontrastwiththatofhomologousCBM13swhererecognitionofcomplexpolysaccharidesresultsfromthecooperativeactionofthreeproteinsub-domainspresentingsimilaraffinities.
IdentificationofaGH62α-L-arabinofuranosidasespecificforarabinoxylanproducedbyPenicilliumchrysogenum.
Sakamoto,T.,Ogura,A.,Inui,M.,Tokuda,S.,Hosokawa,S.,Ihara,H.&Kasai,N.(2011).AppliedMicrobiologyandBiotechnology,90(1),137-146.
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Anarabinoxylanarabinofuranohydrolase(AXS5)waspurifiedfromtheculturefiltrateofPenicilliumchrysogenum31B.ACDNAencodingAXS5(axs5)wasisolatedbyinvitrocloningusingtheN-terminalaminoacidsequenceofthenativeenzymeasastartingpoint.Thededucedaminoacidsequenceoftheaxs5genehashighsimilaritieswiththoseofarabinoxylanarabinofuranohydrolasesofAspergillusniger,Aspergillustubingensis,andAspergillussojae.Modulesequenceanalysisrevealedthata“Glyco_hydro_62”waspresentatposition28–299ofAXS5.Thisisafamilyofα-L-arabinofuranosidaseswhichareallmembersofglycosidehydrolasefamily62.RecombinantAXS5(rAXS5)expressedinEscherichiacoliwashighlyactiveonarabinoxylanbutnotonbranchedsugarbeetarabinan.1H-NMRanalysisrevealedthattherAXS5cleavedarabinosylside-chainslinkedtoC-2andC-3ofsingle-substitutedxyloseresiduesinarabinoxylan.Semi-quantitativeRT-PCRanalysisindicatedthatexpressionoftheaxs5geneinP.chrysogenum31BwasstronglyinducedbyaddingD-xyloseandarabinoxylantotheculturemedium.Moreover,twobindingsitesofXlnR,atranscriptionalactivatorthatregulatestheexpressionofthegenesencodingxylanolyticenzymes,arepresentintheupstreamregionoftheaxs5gene.TheseresultssuggestthatAXS5isinvolvedinxylandegradation.
Endo-1,5-α-L-arabinanasefromaSubseafloorBacillussubtilis:Purification,CharacterizationandNucleotideSequenceofItsGene.
Fukada,Y.,Koide,O.,Miura,T.,Kobayashi,T.,Inoue,A.,&Horikoshi,K.(2011).Journalofappliedglycoscience,58(2),61-66.
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Fourarabinan-degradingenzymesareproducedbyBacillussubtilisJAMA-3-6,whichwasisolatedfromasubseafloorsedimentcorefrom0.5mbelowseaflooratawaterdepthof1,180mofftheShimokitaPeninsulainJapan.Oneoftheenzymes(AbnAF25)waspurifiedfromaculturebroth.Themolecularmassoftheenzymewasaround28kDaasjudgedbySDS-polyacrylamidegelelectrophoresis.TheoptimalpHandtemperaturewerepH6.3and60°Cinphosphatebuffer.AbnAF25degradedwelldebranchedarabinan,lineararabinan,andarabino-oligosaccharaides,butnotarabinoxylan,arabinogalactanorp-nitrophenyl-α-L-arabinofuranoside,whichclassifiestheenzymeasanendo-1,5-α-L-arabinanase.Theendproductsfromlineararabinanweremainlyarabinose,arabinobioseandarabinotriose.ThegeneforAbnAF25wasclonedandsequenced.ThededucedaminoacidsequenceoftheenzymerevealedthehighestsimilaritytothearabinanaseofB.amyloliquefacienswith83%identity.AsAbnAF25didnotshowthedefinitecharacterizationofasubseafloorenzyme,strainJAMA-3-6seemstobeprobablydroppedorco-sedimentedwithasoilcomponent.
AnovelGH43α-L-arabinofuranosidaseofPenicilliumchrysogenumthatpreferentiallydegradessingle-substitutedarabinosylsidechainsinarabinan.
Shinozaki,A.,Kawakami,T.,Hosokawa,S.&Sakamoto,T.(2014).EnzymeandMicrobialTechnology,58,80-86.
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Wepreviouslydescribedthreeα-L-arabinofuranosidases(ABFs)secretedbyPenicilliumchrysogenum31B.Here,wepurifiedafourthABF,termedPcABF43A,fromtheculturefiltrate.Themolecularmassoftheenzymewasestimatedtobe31kDa.PcABF43Ahadthehighestactivityat35°CandataroundpH5.TheenzymeactivitywasstrongonsugarbeetL-arabinanbutweakondebranchedarabinanandarabinoxylan.Lowmolecular-masssubstratessuchasp-nitrophenylα-L-arabinofuranoside,α-1,5-L-arabinooligosaccharides,andbranchedarabinotriosewerehighlyresistanttotheactionofPcABF43A.1H-NMRanalysisrevealedthatPcABF43AhydrolyzedarabinosylsidechainslinkedtoC-2orC-3ofsingle-substitutedarabinoseresiduesinL-arabinan.ReportsconcerningenzymesspecificforL-arabinanarequitelimited.Pcabf43AcDNAencodingPcABF43Awasisolatedbyinvitrocloning.Thededucedaminoacidsequenceoftheenzymeshowshighsimilaritieswiththesequencesofotherfungaluncharacterizedproteins.Semi-quantitativeRT-PCRanalysisindicatedthatthePcabf43AgenewasconstitutivelyexpressedinP.chrysogenum31Batalowlevel,althoughtheexpressionwasinducedwithpecticcomponentssuchasL-arabinose,L-rhamnose,andD-galacturonicacid.AnalysisofenzymaticcharacteristicsofPcABF43A,GH51ABF(AFQ1),andGH54ABF(AFS1)fromP.chrysogenumsuggestedthatPcABF43AandAFS1functionasdebranchingenzymesandAFQ1playsaroleofsaccharificationinthedegradationofL-arabinanbythisfungus.
NascentpectinformedinGolgiapparatusofpeaepicotylsbyadditionofuronicacidshasdifferentpropertiesfromnascentpectinatthestageofgalactanelongation.
ABDel-Massih,R.M.,Rizkallah,H.D.,Al-Din,R.S.,Baydoun,E.A.H.&Brett,C.T.(2007).JournalofPlantPhysiology,164(1),1-10.
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Microsomalmembraneswerepreparedfrometiolatedpea(PisumsativumL.)epicotylsandusedtoformnascent[Uronicacid-14C]pectin.Theenzymeproductswerecharacterizedbyselectiveenzymicdegradation,gelpermeationchromatographyandanalysisofcellulosebindingproperties.Theproductobtainedhadamolecularweightofaround40kDa,whichwassignificantlylowerthanthatofnascent[Gal-14C]pectinpreparedfromthesametissues.Itiscomposedmainlyofpolygalacturonanandperhapsalsorhamnogalacturonan(RG-I).Evidencewasobtainedforthepresenceofaproteinattachedtothenascent[Uronicacid-14C]pectin,butitwasunaffectedbyendoglucanaseanddidnotbindtocellulose.Hence,noxyloglucanappearedtobeattachedtothenascent[Uronicacid-14C]pectin.Amodelisproposedinwhichxyloglucanisattachedtonascentpectinafterformationofhomogalacturonan,butbeforethepectinleavestheGolgiapparatus.
Enzymaticchangesinpecticpolysaccharidesrelatedtothebeneficialeffectofsoakingonbeancookingtime.
Martínez‐Manrique,E.,Jacinto‐Hernández,C.,Garza‐García,R.,Campos,A.,Moreno,E.&Bernal‐Lugo,I.(2011).JournaloftheScienceofFoodandAgriculture,91(13),2394-2398.
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Background:Cookingtimedecreaseswhenbeansaresoakedfirst.However,themolecularbasisofthisdecreaseremainsunclear.Todeterminethemechanismsinvolved,changesinbothpecticpolysaccharidesandcellwallenzymesweremonitoredduringsoaking.Twocultivarsandonebreedinglinewerestudied.Results:Soakingincreasedtheactivityofthecellwallenzymesrhamnogalacturonase,galactanaseandpolygalacturonase.Theiractivityinthecellwallwasdetectedaschangesinchemicalcompositionofpecticpolysaccharides.Rhamnosecontentdecreasedbutgalactoseanduronicacidcontentsincreasedinthepolysaccharidesofsoakedbeans.Adecreaseintheaveragemolecularweightofthepectinfractionwasinducedduringsoaking.Thedecreaseinrhamnoseandthepolygalacturonaseactivitywereassociated(r=0.933,P=0.01,andr=0.725,P=0.01,respectively)withshortercookingtimeaftersoaking.Conclusion:PecticcellwallenzymesareresponsibleforthechangesinrhamnogalacturonanIandpolygalacturonaninducedduringsoakingandconstitutethebiochemicalfactorsthatgivebeancellwallsnewpolysaccharidearrangements.RhamnogalacturonanIisdispersedthroughouttheentirecellwallandinteractswithcelluloseandhemicellulosefibres,resultinginahigherrateofpecticpolysaccharidethermosolubilityand,therefore,ashortercookingtime.
Quantificationoffoodpolysaccharidemixturesby1HNMR.
Merkx,D.W.,Westphal,Y.,vanVelzen,E.J.,Thakoer,K.V.,deRoo,N.&vanDuynhoven,J.P.(2017).CarbohydratePolymers,179,379-385.
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Polysaccharidesarefoodingredientsthatcriticallydeterminerheologicalpropertiesandshelflife.Aqualitativeandquantitativeassessmentonfood-specificpolysaccharidemixturesby1HNMRispresented.Themethodisbasedontheidentificationofintactpolysaccharides,combinedwithaquantitativeanalysisoftheirmonosaccharideconstituents.Identificationofthepolysaccharidesisachievedby1HNMRlineshapefittingwithpurecompoundspectra.ThemonomericcompositionwasdeterminedusingtheSaemanhydrolysisprocedure,followedbydirectmonosaccharidequantificationby1HNMR.Inthequantification,boththemonosaccharidedegradationduringhydrolysis,aswellasacorrectionforthenon-instantaneouspolysaccharidedissolutionweretakenintoaccount.Thesefactorswereparticularlyimportantforthequantificationofpectins.Themethodshowedoverallgoodrepeatability(RSDr=4.1±0.9%)andwithin-laboratoryreproducibility(RSDR=6.1±1.4%)forvariousfoodpolysaccharides.Polysaccharidemixtureswerequantitativelyresolvedbyanon-negativeleastsquaresestimation,usingidentifiedpolysaccharidesandtheirmolarmonosaccharidestoichiometryaspriorknowledge.Theaccuracyandprecisionofthepresentedmethodmakeitapplicabletoawiderangeoffoodpolysaccharidemixtureswithcomplexandoverlapping1HNMRspectra.
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商品咨询
请问哪个公司有检测组织中钙离子浓度的试剂盒 实验动物与组织学...123
简爱刚2021-08-03
我想检测血管组织中的钙离子浓度,不知道哪个公司有试剂盒
下列各组溶液,不加其他试剂就能鉴别的是 初中化学 菁优网123
陌幻念2018-01-30
A,Na2CO3H2SO4HCLNaNO3B,NaOHNaCLMgCL2FeCL3C,HCLAgNO3HNO3MgCL2
D,K2SO4Na2CO3BaCL2NaNO3
D,K2SO4Na2CO3BaCL2NaNO3
CNASCL36:2012《医学实验室质量和能力认可准则在基因扩增检验领域的...123
答案依旧2021-07-22
相关疾病:癫痫请各位老师分享一下,对于红框中的验证要求,你们实验室是怎么验证的呢?对于试剂的批间差异验证,我们是通过留样再测来比对的,想问一下,可不可以使用核酸提取液提取后的DNA冰冻保存好,然后需要比对的时候再拿......
用1mg/mL的铁储备液配制10μg/mL的工作液,用此工作液配制一组标准...123
jinbowen3252018-01-29
例:1。硝酸钡2。硝酸银3。氯化钠4。氯化铜四种溶液检验出的顺序。
通过解答,教会我,谢谢
通过解答,教会我,谢谢
检验科试剂更换记录登记表AU680123
xingfudejueze2021-07-20
最近发现一个问题一般生化仪至少能检测二三十个项目!那么问题是这么多项目要加的试剂R1,R2,算下来也得六七十个瓶瓶罐罐,每天加试剂,倒过来倒过去的,很容易把试剂加错吧?所以就想问一下各位平时工作中有......
Alexa Fluor 488标记亲和纯化山羊抗兔IgG(H+L)二抗 昊鑫生物123
緲酱2018-01-24
不用其他试剂,可以鉴别石蕊、盐酸、氢氧化钙、氢氧化钠、碳酸钠五种溶液,第四个被鉴别出来的物质是( )A.盐酸B.氢氧化钙C.氢氧化钠D.碳酸钠
化学试剂有什么级别以及试剂级别划分的标准是什么 123
fochezhing8672018-03-16
化学试剂的纯度较高,根据纯度及杂质含量的多少,可将其分为以下几个等级。
(1)优级纯试剂 亦称保证试剂,为一级品,纯度高,杂质极少,主要用于精密分析和科学研究,常以GR表示。
(2)分析纯试剂 亦称分析试剂,为二级品,纯度略低于优级纯,杂质含量略高于优级纯,适用于重要分析和一般性研究工作,常以AR表示。
(3)化学纯试剂 为三级品,纯度较分析纯差,但高于实验试剂,适用于工厂、学校一般性的分析工作,常以CP表示。
(4)实验试剂 为四级品,纯度比化学纯差,但比工业品纯度高,主要用于一般化学实验,不能用于分析工作,常以 LR表示。
以上按试剂纯度的分类法已在我国通用。根据化学工业部颁布的“化学试剂包装及标志”的规定,化学试剂的不同等级分别用各种不同的颜色来标志,见表1。
表1 我国化学试剂的等级及标志
(1)优级纯试剂 亦称保证试剂,为一级品,纯度高,杂质极少,主要用于精密分析和科学研究,常以GR表示。
(2)分析纯试剂 亦称分析试剂,为二级品,纯度略低于优级纯,杂质含量略高于优级纯,适用于重要分析和一般性研究工作,常以AR表示。
(3)化学纯试剂 为三级品,纯度较分析纯差,但高于实验试剂,适用于工厂、学校一般性的分析工作,常以CP表示。
(4)实验试剂 为四级品,纯度比化学纯差,但比工业品纯度高,主要用于一般化学实验,不能用于分析工作,常以 LR表示。
以上按试剂纯度的分类法已在我国通用。根据化学工业部颁布的“化学试剂包装及标志”的规定,化学试剂的不同等级分别用各种不同的颜色来标志,见表1。
表1 我国化学试剂的等级及标志
不用任何试剂鉴别物质的方法123
youlovexusong2018-03-26
FeCI3、NaOH、NaCI、HNO3。
不用任何试剂进行物质鉴别的突破口 123
杰少~2018-01-25
1.NaHCO3NaHSO4Ba(NO3)2
2.NaNO3FeCl3AgNO3
分别有什么现象?谢谢回答!
2.NaNO3FeCl3AgNO3
分别有什么现象?谢谢回答!
求化学试剂的其他购买途径!!123
2018-01-28
我家在武汉市远城区,去市区买化学试剂耗时耗力,而且机会也不多。但是我们城区没有试剂店,我又不能网购,求大神给我一些购买方式!尽量方便一些!谢谢!
只用试管和胶头滴管而不用其它试剂无法区分的一组试剂是A.KOH溶液...123
对对对272018-01-30
A.KOH溶液和AlCl3溶液B.Na2CO3溶液和盐酸
C.MgCl2溶液和氨水D.盐酸和NaAlO2溶液
为什么
C.MgCl2溶液和氨水D.盐酸和NaAlO2溶液
为什么
免费发布其他化学试剂求购信息,提供大量求购其他化学试剂产品信...123
sliveryang2021-08-02
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