添加劑輔助變性溶菌酶復性的研究.pdf

1、天津大學碩士學位論文添加劑輔助變性溶菌酶復性的研究姓名：黃艷申請學位級別：碩士專業(yè)：生物化工指導教師：董曉燕20030101ABSTRACTTheroleofvariousdilutionadditiviesontherefoldiagofdenaturedreducedchickeneggwhitelysozymeWaSinvestigated，andeffectoftheconcentrationsofadditives、oxidi

2、zedglutathione(GSSG)、reducedglutathione(GSH)andpH，temperaturewasstudiedtofindproperrefoldingconditionsAkineticmodelwasemployedtoexpresstherenaturationprocessofdenaturedlysozymeinthepresenceofadditivesInthesaruetime，Wedis

3、cussedthepossibilityoflysozymerefoldingbyincreasingadditiviesconcentrationsandreducingguanidiniumchloride(GdmCl)concentrationItwasfoundthattherenaturationyieldWasimprovedinthepresenceofarginine，acetamide，acetone，thiourea

4、andglycerol，andtherewasanoptimaladditiveconcentrationforeachadditive，whichmadetherefoldingteachthehighestyieldTheexprementsresultsshowedthattheoptimalpHandtemperaturewere89and37。C，therationofGSSG：GSHWaSl：1andtheoptimalgl

5、utathioneconcentationwas5mm01／LThekineticmodelWaSingoodagreementwiththeexperimentresultsItWItSfoundthattherefoldingpromotingmechanismofacetamide，thioureaacetone，argininewastheSalrleaSthatofGdmClThatis。witllincreasingaddi

6、tivesconcentrations，therenaturationrateconstantsdecreased，butthereexistedspecialadditivesconcentrationsthatminimizedtheaggregationrateconstants，thusleadingtomaximumrecoveryoftherelativeactivityInthecontraSt，therenaturati

7、onrateconstantsincreasedwithincreasingglycerolconcentration1nthepresenceoflowerGdmClconcentrafion(O06mol／L)，iftheadditivesconcentrations(acetamide，thiourea，acetone，arginine)intherefoldingbufferwereproperlyincreased，there

8、naturationyieldcouldbethesameasthatinthepresenceofhigherGdmClconcentration(02reel／L)HoweverGlycerolWaSdifferenttootheradditivesThatis，increaSingglyeerolconcentrationataproperGdmClconcentrationcouldobtainahighrefoldingyie

9、ldofprotein，F(xiàn)urthermore，therenaturationrecoverydecreasedwhenthelysozymeconcentrationWaSincreaSed，butifweincreasedacetamideconcentration，thedenaturedlysozymeofO8mg／mlcouldbewellrefolded，andtherenaturationyieldwas90％Keywor