lactobacillus helveticus h9全基因组测序及蛋白质组学分析-whole genome sequencing and proteomics analysis of lactobacillus helveticus h9.docx

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2018-05-28 发布于上海
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lactobacillus helveticus h9全基因组测序及蛋白质组学分析-whole genome sequencing and proteomics analysis of lactobacillus helveticus h9.docx

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lactobacillus helveticus h9全基因组测序及蛋白质组学分析-whole genome sequencing and proteomics analysis of lactobacillus helveticus h9

CompleteGenomeSequenceandProteomicAnalysisofLactobacillushelveticusH9AbstractLactobacillushelveticusH9(LABCCIMAU60208)andLactobacillushelveticusH10（LABCCIMAU60227）wereisolatedandidentifiedfromsamplesofTibet,China.ItwaspreservedinKeyLaboratoryofDairyBiotechnologyandEngineering,MinistryofEducation,InnerMongoliaAgriculturalUniversity.Throughscreeningandexperimentalverification,bothLactobacillushelveticusH9andLactobacillushelveticusH10haverelativelyhigherproteolyticactivity,themaindifferencewasthattheskimmedmilkfermentedbystrainH9canproducepeptideswithpotentialantihypertensiveactivity.InordertoexplorethemechanismofLactobacillushelveticusH9fermentedmilkcanproducesubstanceswithACEinhibitoryactivity,weusedwhole-genomesequencingandproteomicsresearchmethodtoanalyze.Twohigh-throughputsequencingmethods,Roche454andIlluminaSolexa,wereusedtodeterminethecompletegenomesequenceofLactobacillushelveticusH9andLactobacillushelveticusH10,usingcomparativegenomicsresearchtostudyandanalyzetheproteolyticsystemoflacticacidbacteria.GenomesequencingresultsshowedthatthegenomeofLactobacillushelveticusH9andthegenomeofLactobacillushelveticusH10moleculesareclosedcircularmolecule,thelengthwere1,871,119bpand2,145,899bprespectively.1,815and2,049ofORFs(openreadingframe).werepredicted.ByCOGfunctionalcategoriesandKEGGdatabases,weanalyzedtheglucosemetabolism,aminoacidmetabolism,proteinhydrolysissystem,nucleicacidmetabolismofstrainH9,andalsoanalyzedtheabilityofexopolysaccharideproduction,CRISPRgenecluster,andinsertionsequence.WeanalyzedtheproteolyticsystemofstrainH9,H10andDPC4571usingcomparativegenomicsstudy,theresultshowedthatthecellwallproteasePrtH,intracellularpeptidasePepN,PepOandPepXmaybethekeyenzymesinthepathwayofthestrainproducedACEinhibitorypeptides.WeusedproteomicsresearchtechniquesandmethodstoconstructthereferencemapsofLactobacillushelveticusH9duringdifferentfermentationstages,analyzetheexpressionofproteinindifferentgrowthstages,thefocusontheproteolyticsystemoflacticacidbacteria.Fromtwo-dimensiona