A Novel Halophilic Lipase, LipBL, Showing High Efficiency in the Production of Eicosapentaenoic Acid (EPA) 英文参考文献.docVIP

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A Novel Halophilic Lipase, LipBL, Showing High Efficiency in the Production of Eicosapentaenoic Acid (EPA) 英文参考文献.doc

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A Novel Halophilic Lipase, LipBL, Showing High Efficiency in the Production of Eicosapentaenoic Acid (EPA) 英文参考文献

ANovelHalophilicLipase,LipBL,ShowingHigh EfficiencyintheProductionofEicosapentaenoicAcid (EPA) DoloresPe′rez1,SaraMart?′n1,GloriaFerna′ndez-Lorente2,MarcoFilice3,Jose′ ManuelGuisa′n3 ,Antonio Ventosa1,Mar?′aTeresaGarc?′a1,Encarnacio′nMellado1* 1Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, Seville, Spain, 2Institute of Industrial Fermentation, Consejo Superior de InvestigacionesCientificas(CSIC),Madrid,Spain,3DepartmentofBiocatalysis,InstituteofCatalysisandPetrochemistry,ConsejoSuperiordeInvestigacionesCientificas (CSIC),AutonomousUniversityCampus,Cantoblanco,Madrid,Spain Abstract Background:Amongextremophiles,halophilesaredefinedasmicroorganismsadaptedtoliveandthriveindiverseextreme saline environments. These extremophilic microorganisms constitute the source of a number of hydrolases with great biotechnologicalapplications.Theinteresttouseextremozymesfromhalophilesinindustrialapplicationsistheirresistance toorganicsolventsandextremetemperatures.MarinobacterlipolyticusSM19isamoderatelyhalophilicbacterium,isolated previouslyfromasalinehabitatinSouthSpain,showinglipolyticactivity. MethodsandFindings:AlipolyticenzymefromthehalophilicbacteriumMarinobacterlipolyticusSM19wasisolated.This enzyme,designatedLipBL,wasexpressedinEscherichiacoli.LipBLisaproteinof404aminoacidswithamolecularmassof 45.3kDaandhighidentitytoclassCb-lactamases.LipBLwaspurifiedandbiochemicallycharacterized.Thetemperaturefor itsmaximalactivitywas80uCandthepHoptimumdeterminedat25uCwas7.0,showingoptimalactivitywithoutsodium chloride,whilemaintaining20%activityinawiderangeofNaClconcentrations.Thisenzymeexhibitedhighactivityagainst short-medium lengthacyl chainsubstrates, althoughitalso hydrolyzesoliveoil andfishoil.The fishoilhydrolysis using LipBLresultsinanenrichmentoffreeeicosapentaenoicacid(EPA),butnotdocosahexaenoicacid(DHA),relativetoitslevels present in fish oil. For improving the stability and to be used in industrial processes LipBL was immob