Evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.

<h4>Background</h4>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 homeostat...

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Autores principales:	Shinji Fukuda, Yumiko Nakanishi, Eisuke Chikayama, Hiroshi Ohno, Tsuneo Hino, Jun Kikuchi
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2009
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/526f30fd6cba42d589ecc2eb47c8c0b0
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spelling	oai:doaj.org-article:526f30fd6cba42d589ecc2eb47c8c0b02021-11-25T06:16:40ZEvaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.1932-620310.1371/journal.pone.0004893https://doaj.org/article/526f30fd6cba42d589ecc2eb47c8c0b02009-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19287504/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>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 reactions to produce several metabolites from undigested food, and those metabolites exert various effects on mammalian cells in a time-dependent manner. We have established a method for the analysis of bacterial metabolic dynamics in real time and used it in combination with statistical NMR procedures.<h4>Methodology/principal findings</h4>We developed a novel method called real-time metabolotyping (RT-MT), which performs sequential (1)H-NMR profiling and two-dimensional (2D) (1)H, (13)C-HSQC (heteronuclear single quantum coherence) profiling during bacterial growth in an NMR tube. The profiles were evaluated with such statistical methods as Z-score analysis, principal components analysis, and time series of statistical TOtal Correlation SpectroScopY (TOCSY). In addition, using 2D (1)H, (13)C-HSQC with the stable isotope labeling technique, we observed the metabolic kinetics of specific biochemical reactions based on time-dependent 2D kinetic profiles. Using these methods, we clarified the pathway for linolenic acid hydrogenation by a gastrointestinal bacterium, Butyrivibrio fibrisolvens. We identified trans11, cis13 conjugated linoleic acid as the intermediate of linolenic acid hydrogenation by B. fibrisolvens, based on the results of (13)C-labeling RT-MT experiments. In addition, we showed that the biohydrogenation of polyunsaturated fatty acids serves as a defense mechanism against their toxic effects.<h4>Conclusions</h4>RT-MT is useful for the characterization of beneficial bacterium that shows potential for use as probiotic by producing bioactive compounds.Shinji FukudaYumiko NakanishiEisuke ChikayamaHiroshi OhnoTsuneo HinoJun KikuchiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 4, Iss 3, p e4893 (2009)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Shinji Fukuda Yumiko Nakanishi Eisuke Chikayama Hiroshi Ohno Tsuneo Hino Jun Kikuchi Evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.
description	<h4>Background</h4>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 reactions to produce several metabolites from undigested food, and those metabolites exert various effects on mammalian cells in a time-dependent manner. We have established a method for the analysis of bacterial metabolic dynamics in real time and used it in combination with statistical NMR procedures.<h4>Methodology/principal findings</h4>We developed a novel method called real-time metabolotyping (RT-MT), which performs sequential (1)H-NMR profiling and two-dimensional (2D) (1)H, (13)C-HSQC (heteronuclear single quantum coherence) profiling during bacterial growth in an NMR tube. The profiles were evaluated with such statistical methods as Z-score analysis, principal components analysis, and time series of statistical TOtal Correlation SpectroScopY (TOCSY). In addition, using 2D (1)H, (13)C-HSQC with the stable isotope labeling technique, we observed the metabolic kinetics of specific biochemical reactions based on time-dependent 2D kinetic profiles. Using these methods, we clarified the pathway for linolenic acid hydrogenation by a gastrointestinal bacterium, Butyrivibrio fibrisolvens. We identified trans11, cis13 conjugated linoleic acid as the intermediate of linolenic acid hydrogenation by B. fibrisolvens, based on the results of (13)C-labeling RT-MT experiments. In addition, we showed that the biohydrogenation of polyunsaturated fatty acids serves as a defense mechanism against their toxic effects.<h4>Conclusions</h4>RT-MT is useful for the characterization of beneficial bacterium that shows potential for use as probiotic by producing bioactive compounds.
format	article
author	Shinji Fukuda Yumiko Nakanishi Eisuke Chikayama Hiroshi Ohno Tsuneo Hino Jun Kikuchi
author_facet	Shinji Fukuda Yumiko Nakanishi Eisuke Chikayama Hiroshi Ohno Tsuneo Hino Jun Kikuchi
author_sort	Shinji Fukuda
title	Evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.
title_short	Evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.
title_full	Evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.
title_fullStr	Evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.
title_full_unstemmed	Evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.
title_sort	evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.
publisher	Public Library of Science (PLoS)
publishDate	2009
url	https://doaj.org/article/526f30fd6cba42d589ecc2eb47c8c0b0
work_keys_str_mv	AT shinjifukuda evaluationandcharacterizationofbacterialmetabolicdynamicswithanovelprofilingtechniquerealtimemetabolotyping AT yumikonakanishi evaluationandcharacterizationofbacterialmetabolicdynamicswithanovelprofilingtechniquerealtimemetabolotyping AT eisukechikayama evaluationandcharacterizationofbacterialmetabolicdynamicswithanovelprofilingtechniquerealtimemetabolotyping AT hiroshiohno evaluationandcharacterizationofbacterialmetabolicdynamicswithanovelprofilingtechniquerealtimemetabolotyping AT tsuneohino evaluationandcharacterizationofbacterialmetabolicdynamicswithanovelprofilingtechniquerealtimemetabolotyping AT junkikuchi evaluationandcharacterizationofbacterialmetabolicdynamicswithanovelprofilingtechniquerealtimemetabolotyping
_version_	1718413981959323648

Evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping.

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