a fluorosensor for two-dimensional measurements of extracellular enzyme activity

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1、MarineChemistry123(2011)23–31ContentslistsavailableatScienceDirectMarineChemistryjournalhomepage:www.elsevier.com/locate/marchemAfluorosensorfortwo-dimensionalmeasurementsofextracellularenzymeactivityinmarinesedimentsZhenruiCao,QingzhiZhu,RobertC.Aller⁎,JosephineY.AllerSchoolofMarineandAtmosphericS

2、ciences,StonyBrookUniversity,StonyBrook,NY11794-5000,UnitedStatesarticleinfoabstractArticlehistory:Anovelplanarfluorosensorhasbeendevelopedformeasuringtwo-dimensionalactivitydistributionsoftheReceived22May2010extracellularenzyme,leucine-aminopetidase(Leu-AP),inmarinesediments.Thesensorisafoilconsis

3、tingofReceivedinrevisedform24August2010anenzymesubstrateL-leucine7-amido-4-methylcoumarinhydrochloride(Leu-MCA)andaHydromedD4Accepted8September2010membrane,backedbyapolyestersheet.Initspresentapplication,thesensorfoilisintroducedintosurficialAvailableonline18September2010depositstoadepthof~10–15cm,

4、andtimedependentfluorescenceproductionoccurringinsuccessivedigitalimagesisusedtoresolveenzymeactivitypatterns.Correctionsformicroenvironmentalvariability(e.g.,Keywords:diffusionandadsorptionproperties)andbackgroundreflectanceweremade.TheplanarfluorosensorEnzymeplanarfluorosensorExtracellularenzymeacti

5、vitytechniquewasstandardizedbycomparisonwithtraditionalslurrybasedmethodsusinghomogenizedLeucine-aminopetidasesedimentsections.Spatiallyaveraged,verticalenzymeactivityprofilesmeasuredusingtheimagingsensorTwo-dimensionalmeasurementssystemwerecontrastedwiththetraditionalmethodtoverifythereliabilityof

6、thisnewapproach.ThenewMarinesediments2systemresolvesenzymeactivityonasubmillimeterscaleoverrelativelylargeareas(~50–100cm).Inadditiontoallowingrapiddeterminationofenzymeactivitypatternsinundisturbedmaterial,thetransparentnatureofthesensingfoilspermitsdirectcorrelationofenzymedistributionstoheterog

7、eneousbiologicalandphysicalstructure.©2010ElsevierB.V.Allrightsreserved.1.Introductionheterogeneityinenzymeactivityisobscuredandthepossibleassociationofactivitypatternswithsedimentarystructureparticu-Belowthephot