天然木质纤维素降解相关酶的宏蛋白质组学分析与鉴定

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1、福建师范大学吕睿瑞硕士学位论文的胞外蛋白样品通过二维凝胶电泳分离各个蛋白点，对所有的可分辨点进行MODIL．TOF．TOF串联质谱鉴定，最终鉴定出5木质纤维素降解酶和葡聚糖降解酶，有过氧化氢酶、葡萄糖淀粉酶、葡聚糖1，4-a．葡萄糖苷酶、13．1，3．内切葡聚糖酶、1，4-13．D．葡聚糖纤维二糖水解酶，它们之间的比例约为5：30：20：40：10。我们的研究首次表明了真正参与天然木质纤维素降解过程中涉及酶的真实情况。随着研究的深入，一个完整的酶档案将为将来构建高效的木质纤维素降解复合酶系提供理论基础。关键词

2、：土壤，木质纤维素降解机制，酶，微生物，宏蛋白质组学，复合酶系AbstractAbstractNaturallignocellulosicmaterialsarewidelyavailableforthemicroorganismsintheirhabitats．Thesemicroorganismsproduceabatteryofenzymesandworksynergicallytobiodegradelignocellulose．Therefore，totrulyunderstandhowmicroo

3、rganismshydrolyzethelignocellulose，theymustbestudiedassystemsinnatureratherthanindividualorafewatatime。Inthispaper，theapplicationofmetaproteomicstounderstandmicrobiallignocelluloseutilizationstrategieswaswelldescribed．Soilsamplesfromdifferentclimateandveget

4、ationtypeswerecollected．Celluloseandxylanaseactivityandtheefficiencyoflignocellulosedegradationweredeterminedsubsequently．Theresultshowedthesoilsamplehadahigherxylanaseactivitybutamuchlowerevenzeroincelluloseactivity．Thehighestxylanaseactivityreachedupto192

5、．92u／g．Afterenrichmentusingnaturallignocelluloseasculturemateria，theemzymesactivitiesofmostsoilsampleswereincreased．Thehighestcelluloseandxylanaseactivitywere6530．99u／gand1275．16u／g，respectively．Finally,HN一5soilsamplewasselectedasthenextexperimentaltargetby

6、consideratingthreeindicatorscomprehensively．Theproteomicsapproachwasbuiltbasedonalignocellulose—degradingAspergillusnigerstrainHCB一3isolatedfromHN-5soilsample．AftermediumoptimizationforstrainHCB一3toimprovetheproductionoflignocellulose—degradingenzymes，theto

7、talsolubleextracellularproteinswereextractedfromHCB一3brothandappliedinproteomicsanalysis．7hemicellulose—degradingenzymeswereidentificatedfromthe2Dgel，as3xylanase，1arabinoxylandegradingenzyme，1a-L—arabinofuranosidase，1仅一glucuronidaseand1feruloylesterase．Furt

8、herresultsaboutratiooftheseemzymesobtainedusingthesoftwareofImageMaster2DPlatinum．ThesuccessfulanalysisconfirmedthefeasibilityoftheentireproteomicstrategyinIII’‘一，，l啊“¨、r一^一如：福建师范大学吕睿瑞硕士学位论文identifying