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Open AccessArticle

Novel Strategy for Non-Targeted Isotope-Assisted Metabolomics by Means of Metabolic Turnover and Multivariate Analysis

1
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan
2
RIKEN Center for Sustainable Resource Science, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan
*
Author to whom correspondence should be addressed.
Metabolites 2014, 4(3), 722-739; https://doi.org/10.3390/metabo4030722
Received: 21 April 2014 / Revised: 5 August 2014 / Accepted: 12 August 2014 / Published: 25 August 2014
(This article belongs to the Special Issue Metabolomics and Biotechnology)
Isotope-labeling is a useful technique for understanding cellular metabolism. Recent advances in metabolomics have extended the capability of isotope-assisted studies to reveal global metabolism. For instance, isotope-assisted metabolomics technology has enabled the mapping of a global metabolic network, estimation of flux at branch points of metabolic pathways, and assignment of elemental formulas to unknown metabolites. Furthermore, some data processing tools have been developed to apply these techniques to a non-targeted approach, which plays an important role in revealing unknown or unexpected metabolism. However, data collection and integration strategies for non-targeted isotope-assisted metabolomics have not been established. Therefore, a systematic approach is proposed to elucidate metabolic dynamics without targeting pathways by means of time-resolved isotope tracking, i.e., “metabolic turnover analysis”, as well as multivariate analysis. We applied this approach to study the metabolic dynamics in amino acid perturbation of Saccharomyces cerevisiae. In metabolic turnover analysis, 69 peaks including 35 unidentified peaks were investigated. Multivariate analysis of metabolic turnover successfully detected a pathway known to be inhibited by amino acid perturbation. In addition, our strategy enabled identification of unknown peaks putatively related to the perturbation. View Full-Text
Keywords: metabolomics; stable isotope; isotope-assisted metabolomics; metabolic turnover analysis; non-targeted analysis; gas chromatography; mass spectrometry; Saccharomyces cerevisiae metabolomics; stable isotope; isotope-assisted metabolomics; metabolic turnover analysis; non-targeted analysis; gas chromatography; mass spectrometry; Saccharomyces cerevisiae
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Nakayama, Y.; Tamada, Y.; Tsugawa, H.; Bamba, T.; Fukusaki, E. Novel Strategy for Non-Targeted Isotope-Assisted Metabolomics by Means of Metabolic Turnover and Multivariate Analysis. Metabolites 2014, 4, 722-739.

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