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《產(chǎn)朊假絲酵母尿酸酶的純化、特性研究及其基因擴增》由會員上傳分享，免費在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、重慶醫(yī)科大學(xué)碩士研究生學(xué)位論文PREPARATIoN、CHARACTERIZATIONANDGENICCLONINGoFTHEURlCASEFROMCAⅣD刪Um坶ABSTRACTUricacidastheend—productofpurinemetabolisminhumanbodyispassedoutthroughkidney．Iftheproductionofuricacidsurpassesthatexcretedbykidney,theconcentrationofserumuricacidwil
2、lbeincreasedandhyperuricemiawillOCCULT．Hyperuricemiaisassociatedwiththedevelopmentandprogressofgout，cardiovasculardiseases，tumorlysissyndromeandrenaldisease．Soitisnecessarytodevelopefficientdrugforhyperuricemia．Currently,theuricaseisconsideredtobeanidealdru
3、gtotreathyperuricemia．Herein，theCandidautilis(C．GM．C．C．2．1008)anditsuricasewasinvestigated．1．TheincubationoftheCandidautilis(C．GM．C．C．2．1008)andpreparationandpurificationoftheuricase．Afterbeinginducedbyuricacid，Thecellswereharvestedthroughcentrifugationandb
4、rokenwithultrasound．Thelysateisfractionedwithammoniumsulfateandtheuricasespecificactivityoftheproductreachedtomorethan0．32U／mgprotein．DuringtheDEAE—cellulosechromatography,theuricasecouldn’tbeabsorbedbutitsspecificactivityreachedto7．1U／ragaftertwiceDEAE—cel
5、lulosechromatography．2．Thecharacterizationofuricase．4重慶醫(yī)科大學(xué)碩士研究生學(xué)位論文Thenativeuricasewas134．0kDbySephadexG一200gelfiltration．AftertwicechromatographyonDEAE-cellulose52，thespecificactivityofthepartlypurifieduricasewasmorethan7．0U／mgprotein．Onlysinglepolypeptid
6、eabout33．0kDwasresolvedbySDS—PAGEwiththissample．Matrix··assistedlaserdesorptionionizationtime-·of-flightMSanalysisresolveda67．7kDpolypeptideastheprincipalcomponentanda131．4kDpolypeptideofslightlylowerabundance，butnosinglepolypeptidewasdetectedwiththesamesam
7、ple．TheoptimumpHforthisuricasewascloseto8．8at25+0．5℃．Thisuricasewasstableat37℃for4handmorethan50％activitywasreservedatpH7．4．TheMichaelis—MentenconstantWas(32．8+3．1)IJmol／L(n=10)．Exceptthatadenosine、guanosineandinosinecouldn’tinhibittheuricase，theinhibitionc
8、onstantofOteracilpotassium、xanthine，guanine，hypoxanthine、adenineandallopurinolwasindividually(1．1+0．1)“mol／L(以=3)、(4．84-0．2)gmol／L(刀=3)、(37．94-4．2)gmol／L(療23)、(500．0-3：60．O)lmaol／L∽=3)、(748．94-82．O)lxr
