Evaluation and Characterization of Bacterial Metabolic Dynamics with a Novel Profiling Technique, Real-Time Metabolotyping 英文参考文献.docVIP

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Evaluation and Characterization of Bacterial Metabolic Dynamics with a Novel Profiling Technique, Real-Time Metabolotyping 英文参考文献.doc

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Evaluation and Characterization of Bacterial Metabolic Dynamics with a Novel Profiling Technique, Real-Time Metabolotyping 英文参考文献

EvaluationandCharacterizationofBacterialMetabolic DynamicswithaNovelProfilingTechnique,Real-Time Metabolotyping ShinjiFukuda1,2,YumikoNakanishi2,3,EisukeChikayama3,HiroshiOhno1,2,TsuneoHino4* ,Jun Kikuchi2,3,5 * 1RIKENResearchCenterforAllergyandImmunology,Suehiro-cho,Yokohama,Japan,2InternationalGraduateSchoolofArtsandSciences,YokohamaCityUniversity, Suehiro-cho,Yokohama,Japan,3RIKENPlantScienceCenter,Suehiro-cho,Yokohama,Japan,4DepartmentofLifeScience,MeijiUniversity,Tama-ku,Kawasaki,Japan, 5GraduateSchoolofBioagriculturalSciences,NagoyaUniversity,Furo-cho,Nagoya,Japan Abstract Background: Environmental processes in ecosystems are dynamically altered by several metabolic responses in microorganisms, including intracellular sensing and pumping, battle for survival, and supply of or competition for nutrients. Notably, intestinal bacteria maintain homeostatic balance in mammals via multiple dynamic biochemical reactionstoproduceseveralmetabolitesfromundigestedfood,andthosemetabolitesexertvariouseffectsonmammalian cellsinatime-dependentmanner.Wehaveestablishedamethodfortheanalysisofbacterialmetabolicdynamicsinreal timeanduseditincombinationwithstatisticalNMRprocedures. Methodology/PrincipalFindings:Wedevelopedanovelmethodcalledreal-timemetabolotyping(RT-MT),whichperforms sequential 1H-NMRprofilingandtwo-dimensional(2D) 1H, 13C-HSQC(heteronuclearsinglequantumcoherence)profiling during bacterial growth in an NMR tube. The profiles were evaluated with such statistical methods as Z-score analysis, principalcomponentsanalysis,andtimeseriesofstatisticalTOtalCorrelationSpectroScopY(TOCSY).Inaddition,using2D 1H, 13C-HSQC with the stable isotope labeling technique, we observed the metabolic kinetics of specific biochemical reactions based on time-dependent2D kineticprofiles. Usingthese methods,we clarifiedthepathway forlinolenic acid hydrogenationbyagastrointestinalbacterium,Butyrivibriofibrisolvens.Weidentifiedtrans11,cis13conjugatedlinoleicacid as the intermediate of l