Molecular cloning and functional expression of a novel extracellular lipase from the thermotolerant yeastCandida thermophila英文资料

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1、MolecularcloningandfunctionalexpressionofanovelextracellularlipasefromthethermotolerantyeastCandidathermophilaJantapornThongekkaew&ChuenchitBoonchirdDepartmentofBiotechnology,FacultyofScience,MahidolUniversity,Bangkok,ThailandCorrespondence:ChuenchitBoonchird,AbstractDepartmentofBio

2、technology,FacultyofThethermotolerantyeastCandidathermophilaSRY-09isolatedfromThailandScience,MahidolUniversity,RamaVIRoad,Phayathai,Ratchathewi,Bangkok10400,producesanextracellularlipasethathydrolysesvarioustriglycerides.ToclonetheThailand.Tel.:166022015304;fax:1660geneencodingthel

3、ipase,Saccharomycescerevisiaewastransformedwitha23547160;e-mail:sccbc@mahidol.ac.thC.thermophilagenomiclibraryandscreenedforlipaseactivityonmediumcontainingoliveoilemulsionandrhodamineB.OneC.thermophilalipasegeneReceived1May2006;revised1September(CtLIP)wasfoundthatcontainedanORFof13

4、17bpencodingadeduced2006;accepted4September2006.polypeptideof438aminoacids.CandidathermophilalipasecontainedaGly-Asp-Firstpublishedonline21November2006.Ser-Gln-GlymotifwhichmatchedtheconsensusGly-X-Ser-X-Glyconservedamonglipolyticenzymes.HeterologousexpressionoftheclonedCtLIPunderth

5、eDOI:10.1111/j.1567-1364.2006.00173.xcontrolofthealcoholoxidasegene(AOX1)promoterinthemethylotrophicyeastPichiapastoris,andenzymaticmeasurementsconfirmedthefunctionoftheEditor:GuentherDaumrespectiveproteinasalipase.TherecombinantCtLIPcouldhydrolysevariousKeywordssubstratesathightempe

6、rature(551C)withhigherefficiencythanat37or451CCandidathermophila;extracellularlipase;andpreferentiallyhydrolysedtwo-positionalesterbonds.AswithC.thermophila,Saccharomycescerevisiae;heterologoustheheterologouslyexpressedlipasewassecretedintothemediumbyPichiapastoris.expression;Pichiap

7、astoris.digestionfornutrientacquisition.TheseenzymesmightIntroductionhelpamicroorganismtogrowincarbohydrate-restrictedLipasesortriacylglycerolhydrolase(E.C.3.1.1.3)catalysetheenvironmentsorenvironmentswherelipidsarethesolehydrolysisandsynthesisofrelativelylong-chainacylglycerolscarb

8、onsource(Stehretal.