微生物代谢组学的前处理及分析技术

微生物学通报DEC20,2009,36(12):1882~1887Microbiology©2009byInstituteofMicrobiology,[e-mail]基金项目：(No.06ZR14112)*通讯作者：Tel:[phone];:[e-mail]收稿日期：2009-04-30;接受日期：2009-06-30专论与综述微生物代谢组学的前处理及分析技术董玲玲柴逸峰曹颖瑛朱臻宇*(200433)摘要:微生物代谢组学主要研究细胞生长或生长周期某一时刻细胞内外所有低分子量代谢物。分析技术的不断发展促进了微生物代谢组学研究的进展。本文结合微生物样品前处理方法,综述了目前研究中所采用的各种分析技术的特点与应用,并展望微生物代谢组学研究中分析技术的发展趋势。关键词:分析技术,样品前处理,微生物代谢组学PreparationandAnalyticalMethodintheStudyofMicrbialMetabolomicsDONGLing-LingCHAIYi-FengCAOYing-YingZHUZhen-Yu*(SchoolofPharmacy,SecondMilitaryMedicalUniversity,Shanghai200433,China)Abstract:Microbialmetabolomicsisasubjectthatchieflystudyingallthelowmolecularweightmetabo-litesinanorganismorcellsduringtheirgrowingprocess.Theprogressofanalyticaltechnologypromotesmicrobialmetabolomicstomakeadvancement.Inthispaper,thecommonlyusedanalyticaltechnology,samplepreparationanditsapplicationwerediscussedandtheprospectsoftheanalyticalmethodswerealsodiscussed.Keywords:Analyticaltechnology,Samplepreparation,Microbialmetabolomics,,,,,ElmrothGC-MS[1],[2][3][4,5][4,6,7][8,9],,:,,(NMR)(MS),:1883[10],(GC-MS)(CE-MS),DIMS[11],1前处理技术,,MashegoMR,,[12,13]1.1淬灭,−40°C80°C,,−150°C,,0°C()(),,,,,,,,[14],,,,1.2细胞内代谢产物的提取,,[13]MaharjanRP-6,,,(−40°C),[15],,,42(HEPES)[16](PIPES)[14],,,2分析技术平台1[17]-2.1NMR(Nuclearmagneticresonance,NMR),,NMR,1H()-,H,1H-,;13C-,,NMR,,NMR[18,19],NMR,NMR,,,950MHz,[10](1884微生物学通报2009,Vol.36,No.12图1代谢组学研究中常用的技术平台Fig.1AnalysisplatforminmetabolomicsMHz,MHz),-NMR(2D-NMR),,1H-13C-,,2D1D-Ulocladiumatrumcyclopeptolide1,cyclopeptolide1,,cyclopeptolide1,[20]NMR,,[21]2.2MS,2.2.1DIMS:(DIMS),Allen,250(m/z),,,,,,[11]DIMS,,,DIMS,,,,(m/z),,,GeorginaDIMS(Q-TOF)GC−TOF−MS[22],,Sandrine:1885(GC-MS),DIMS,,,[22,23]2.2.2GC-MS:(Gaschroma-tography-massspectrometer,GC-MS),,,,[24]GC-MS,[14,25−27],SteveO’Hagan-(GC-TOF/MS),,[7],Villas-BôasSG,[14],[28],[23],ClostridiumproteoclasticumB316T[29],GC-MS,,,,,,2.2.3HPLC/UPLC-MS:-(Liquidchromatography-massspectrometer,LC-MS)(LC),,(MS)(MS/MS)NMR,LC-MS;GC-MS,,,,,,(Ultraper-formanceliquidchromatography,UPLC),[30]YuBJnano-HPLC-MS/MS,,(Tricarboxylicacid,TCA)[31]JosephJDallugeLC-MS-MS20,,[32]2.2.4CE-MS:(CE-MS),,[33],,HaradaCE-MS[34],CE-MS,2.3光谱法及其它方法NMRMS,,[35];13CNMRMS[36]3展望,,,,,,,,,参考文献[1]ElmrothI,SundinP,ValeurA,etal.Evaluationofchro-1886微生物学通报2009,Vol.36,No.12(3):215−228.[2]BundyJG,WilleyTL,CastelRS,etal.DiscriminationofpathogenicclinicalisolatesandlaboratorystrainsofBa-cilluscereusbyNMR-basedmetabolomicprofiling.FEMSMicrobiolLett,2005,242(1):127−136.[3]RuamsdonkLM,TeusinkB,BroadhurstD,etal.Afunc-tionalgenomicsstrategythatusesmetabolomedatatore-vealthephenotypeofsilentmutations.NatBiotechnol,2001,19(1):45−50.[4]BuchholzA,HudebausJ,WandreyC,etal.Metabolomics:quantificationofintracellularmetabolitedynamics.BiomolEng,2002,19(1):5−15.[5]DaunerM,BaileyJE,SauerU.MetabolicfluxanalysiswithacomprehensiveisotopomermodelinBacillussub-tilis.BiotechnolBioeng,2001,76(2):144−156.[6]DallugeJJ,SmithS,Sanchez-RieraF,etal.Potentialoffermentationprofilingviarapidmeasurementofaminoacidmetabolismbyliquidchromatography-tandemmassspectrometry.JChromatogrA,2004,1043(1):3−7.[7]O’HaganS,DunnWB,BrownM,etal.Closed-loop,mul-tiobjectiveoptimizationofanalyticalinstrumentation:gaschromatography/time-of-flightmassspectrometryofthemetabolomesofhumanserumandofyeastfermentations.AnalChem,2005,77(1):290−303.[8]BoersmaMG,SolyanikovaIP,vanBerkelWJH,etal.19FNMRmetabolomicsfortheelucidationofmicrobialdeg-radationpathwaysoffluorophenols.JIndMicrobiolBio-technol,2001,26(1/2):22−34.[9]NarasimhanK,BasheerC,BajicVB,etal.Enhancementofplant-microbeinteractionsusingarhizosphereme-tabolomics-drivenapproachanditsapplicationinthere-movalofpolychlorinatedbiphenyls.PlantPhysiol,2003,132(1):146−153.[10]LindonJC,NicholsonJK.Analyticaltechnologiesformetabonomicsandmetabolomics,andmulti-omicinfor-mationrecovery.TrendsinAnalyticalChemistry,2008,27(3):194−204.[11]AllenJ,DaveyHM,BroadhurstD,etal.Discriminationofmodesofactionofantifungalsubstancesbyuseofmetabolicfootprinting.AppliedandEnvironmentalMi-crobiology,2004,70(10):6157−6165.[12]MashegoMR,RumboldK,DeMeyM,etal.Microbialmetabolomics:past,presentandfuturemethodologies.BiotechnolLett,2007,29(1):1−16.[13],..,2008,6:404−407.[14]Villas-BôasSG,Højer-PedersenJ,AkessonM,etal.Globalmetaboliteanalysisofyeast:evaluationofsamplepreparationmethods.Yeast,2005,22(14):1155−1169.[15]MaharjanRP,FerenciT.Globalmetaboliteanalysis:theinfluenceofextractionmethodologyonmetabolomepro-filesofEscherichiacoli.AnalBiochem,2003,313(1):145−154.[16]OldigesM,KunzeM,DegenringD,etal.Stimulation,monitoring,andanalysisofpathwaydynamicsbymeta-bolicprofilinginthearomaticaminoacidpathway.Bio-technolProg,2004,20(6):1623−1633.[17]GoodacreR,VaidyanathanS,DunnWB,etal.Me-tabolomicsbynumbers:acquiringandunderstandingglobalmetabol