靈芝漆酶及部分胞外酶研究.pdf

1、北京協(xié)和醫(yī)學院中國醫(yī)學科學院碩士學位論文靈芝漆酶及部分胞外酶研究姓名：劉健鵬申請學位級別：碩士專業(yè)：生藥學指導教師：蘭進20090501Pekingunionmedicalcollege(：盤i絲雖￡星4￡鯉旦壘絲理2f絲壘型i￡旦』點坌!堡絲￡l望△墜￡旦魚!AbStHactGanodermalucidumisafamousfungiwithveryhighmedicinalvalueinChinaTheextracellularen

2、zymesinvolvedintheprocesstogetnutritionforGlucidumwhencultured，wererelatedtoitsoutputandqualityAsoneofextracellularenzymes，laccasefromGllucidumisalsoamultifunctionphenoloxidasewithwideapplicationInordertoelucidatetherela

3、tionbetweenextracellularenzymesandoutputandqualityofG1ucidumandpromotetheapplicationofG1ucidumlaccasewestudiedthevariationofextracellularenzymesandpolysaccharidesofG1ucidumS3culturedAfterscreeninginducersoflaccase，wealso

4、usedaprocedurewhichinvolvedammoniumsulfateprecipitation，DEAE—sepharoseanionexchangechromatographyandSource15PHEhydrophobicchromatographytopurifylaccaseandthencharacterizeditindetailItwasshowedthattheligninmodifyingenzyme

5、sofG1ucidumgrownwithcottonseedskinaSthesolecarbonandenergysourceconsistedoflaccaseandmanganesedependentperoxidase，noligninperoxidasewasobservedEveryenzymehadadifferentvariationtrendCulturemethodshadaneffectonthepolysacch

6、aridecontentThemaximummyceliumbiomassappearedonthe7thday，whilethemaximumactivityofthelaccaseenzymeappearedonthe9thdayThemajorityofthepotentialtestedwereabletoserveasinducersforlaccasetosomeextent，amongwhich26dimethylanil

7、ineWasfoundtobethebestwhentheconcentrationWas08mmol／LWiththenative—PAGEzymogram，itWasinferredthatG1ucidumsecretedonlyasinglelaccaseinliquidfermentation，notaffectedbyinducersThepurifiedlaccaseWasasinglesubmitproteinwitham

8、olecularweightof596KDabySDSPAGETheKmwas902x10～mol／LwithABTSassubstrate，while108x10。mol／LwithguaiacolassubstrateThislaccasewasstablefrompH60to80Thetemperatureoptimumforitsactivitywas45。CTheresearchalsoshowedthatalotofcomp