Pectinaseassay Pectinasesareactuallyamixtureofenzymes,which,alongwithotherssuchascellulase,arewidelyusedinthefruitjuiceindustrywheretheyarewidelyusedtohelpextract,clarifyandmodifyfruitjuices.(Seethefootofthispageformoreinformation) Pectinsarelargepolysaccharidemolecules,madeup(mainly)ofchainsofseveralhundredgalacturonicacidresidues.Enzymesinthispectinasegroupincludepolygalacturonases,pectinmethylesteraseandpectinlyases.Thesepectinaseenzymesactindifferentwaysonthepectins,whicharefoundintheprimarycellwallsandinthemiddlelamella.PectinsarewellknownalsofortheirABIlitytoformgels. Pectinasesareproducedduringthenaturalripeningprocessofsomefruits,wheretogetherwithcellulases,theyhelptosoftentheircellwalls.TheseenzymesarealsosecretedbyplantpathogenssuchasthefungusMoniliniafructigenaandthesoft-rotbacteriumErwiniacarotovora,aspartoftheirstrategyforpenetratingtheplanthostcellwalls.Infact,theproductsofsuchenzymeassaults(oligosaccharins)actasasignalwhichinducesuninfectedcellstodefendthemselves. Theprincipleofthisassaydependsuponmeasuringtheamountofwateryjuicereleasedfromtinnedapplepuree(whichisveryrichinpectin)asaresultofpectinaseaction. Tinnedorbottledapplepuree(sauce)canbepurchasedforthisassay. Thedifferencesinthevolumeofjuicebetweenthetwotubesgivesameasureofthepectinaseactivityintheextract. Ifthisexperimentisbeingcarriedoutonalargescale,theinitialdrainagefromthefunnelscouldbecollectedintesttubes,andthenthevolumeofjuicecollectedismeasuredlater. Manyindustrialenzymeproductsaremixturesofdifferentenzymes.Thusa"pectinase"preparationmightcontainarangeofpectinasesandcellulases.Otherpreparationsmightcontainonlyasingletypeofenzyme,especiallyiftheenzymesareproducedbygenetically-modifiedstrainsorarehighlypurified. Whenplanningyourowninvestigations,itisthereforeveryimportanttostudythedatasheetsuppliedwitheachenzyme.Thiswillindicatewhethertheproductisamixtureorcontainsjustonetypeofenzyme. Thedatasheetalsoprovidesaroughguidetohowtheenzymemightbehave.Inpractice,however,enzymeactivityisaffectedbymanythings(e.g.pH,temperature,thepresenceofinhibitorsorcofactors)whichwillaffecttheresultsyouobtain.Inaddition,specimenactivitygraphsareoftenpreparedusingsimplesubstancesunderidealconditionsratherthanthecomplexsubstratesandsub-optimalconditionsthatmaybeencounteredinanindustrialorschoolcontext. Moreinformation:-InajamandoutofjuiceAnNCBE/Unilevereducationalguidetoenzymesinfruitjuiceproduction
Note:whenusingcommerciallypreparedenzymesinyourinvestigations...