预览加载中，请您耐心等待几秒...

毕赤酵母表达MSL的发酵条件研究.docx

预览
1/2
2/2

在线预览结束，喜欢就下载吧，查找使用更方便

下载提示 文本预览

如果您无法下载资料，请参考说明：

1、部分资料下载需要金币，请确保您的账户上有足够的金币

2、已购买过的文档，再次下载不重复扣费

3、资料包下载后请先用软件解压，在使用对应软件打开

毕赤酵母表达MSL的发酵条件研究 Title:FermentationConditionsfortheExpressionofMSLinPichiapastoris Abstract: TheaimofthisstudyistoinvestigatetheoptimalfermentationconditionsfortheexpressionofMSL(mammaliansmallmoleculeligand)inPichiapastorisusingarecombinantbiotechnologicalapproach.Theoptimizationoffermentationparameterssuchastemperature,pH,carbonandnitrogensources,andinducerconcentrationwereevaluatedtoenhancetheexpressionlevelsofMSL.TheresultsofthisstudyprovideinsightsintotheefficientproductionofMSLusingPichiapastorisasahostorganisminbiotechnologicalapplications. I.Introduction Theexpressionofrecombinantproteinsinmicrobialsystemshasrevolutionizedthefieldofbiotechnology,offeringvariousadvantagesovertraditionalmethods.Pichiapastorishasemergedasanattractivemicrobialhostduetoitsabilitytoefficientlyproduceheterologousproteinswithproperfoldingandpost-translationalmodifications.ThisstudyfocusesontheoptimizationoffermentationconditionsfortheexpressionofMSL,amammaliansmallmoleculeligand,inPichiapastoris. II.MaterialsandMethods ArecombinantPichiapastorisstrainexpressingthetargetMSLgenewasconstructed.Fermentationconditionsincludingtemperature,pH,carbonandnitrogensources,andinducerconcentrationwereoptimizedusingatwo-levelfullfactorialdesign.ThegrowthcharacteristicsofPichiapastorisweremonitoredbymeasuringcelldensityandbiomass.TheexpressionofMSLwasassessedbySDS-PAGEandWesternblottinganalyses. III.Results TheoptimalfermentationconditionsfortheexpressionofMSLinPichiapastorisweredeterminedasfollows: -Temperature:30°C -pH:6.0 -Carbonsource:glycerol -Nitrogensource:peptone -Inducerconcentration:0.5%methanol Undertheseconditions,Pichiapastorisexhibitedenhancedgrowthrates,attaininghigherbiomasslevelsincomparisontootherconditions.TheexpressionofMSLwasalsosignificantlyincreased,asevidencedbythepresenceofthedesiredproteinbandonSDS-PAGEgelsandconfirmedbyWesternblotting. IV.Discussion Temperatureplaysacrucialroleinproteinexpressionasitaffectsthegrowthandmetabolismofthehostorganism.Inthisstudy,atemperatureof