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Optimised protocols for the metabolic profiling of S. cerevisiae by 1H-NMR and HRMAS spectroscopy

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Abstract

An optimised extraction protocol for the analysis of Saccharomyces cerevisiae aqueous and organic metabolites by nuclear magnetic resonance spectroscopy that allows the identification and quantification of up to 50 different compounds is presented. The method was compared with other metabolic profiling protocols for S. cerevisiae, where generally different analytical techniques are applied for metabolite quantification. In addition, the analysis of intact S. cerevisiae cells by HRMAS was implemented for the first time as a complementary method. The optimised protocols were applied to study the metabolic effect of glucose and galactose on S. cerevisiae growth. Furthermore, the metabolic reaction of S. cerevisiae to osmotic stress has been studied.

Graphical description of the metabolic profiling study carried out on S. cerevisiae

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Acknowledgements

This work has been supported by grants from the MINECO (BFU2011-23418 and SAF2011-28350), GV (PROMETEO/2012/061 and ACOMP2013/104) and the Centro de Investigación Príncipe Felipe. M.M. Molina-Navarro has been supported by a Juan de la Cierva Contract (MEC).

The authors wish to thank Dr. Emilia Matallana for critical comments on the manuscript, Dr. Proft for technical advice and A. Llopis and E. Barber for technical support.

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Authors and Affiliations

  1. Gene Expression coupled to RNA Transport Laboratory, Centro de Investigación Príncipe Felipe (CIPF), C. Eduardo Primo Yúfera 3, 46012, Valencia, Spain

    Maria Micaela Molina-Navarro, Marta Tormos-Pérez & Susana Rodríguez-Navarro

  2. Nuclear Magnetic Resonance Service, Centro de Investigación Príncipe Felipe (CIPF), C. Eduardo Primo Yúfera 3, 46012, Valencia, Spain

    Martina Palomino-Schätzlein

  3. Structural Biochemistry Laboratory, Centro de Investigación Príncipe Felipe (CIPF), C. Eduardo Primo Yúfera 3, 46012, Valencia, Spain

    Antonio Pineda-Lucena

Authors
  1. Martina Palomino-Schätzlein
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  2. Maria Micaela Molina-Navarro
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  3. Marta Tormos-Pérez
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  4. Susana Rodríguez-Navarro
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  5. Antonio Pineda-Lucena
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Corresponding authors

Correspondence to Susana Rodríguez-Navarro or Antonio Pineda-Lucena.

Additional information

Martina Palomino-Schätzlein and Maria Micaela Molina-Navarro contributed equally to this work.

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Palomino-Schätzlein, M., Molina-Navarro, M.M., Tormos-Pérez, M. et al. Optimised protocols for the metabolic profiling of S. cerevisiae by 1H-NMR and HRMAS spectroscopy. Anal Bioanal Chem 405, 8431–8441 (2013). https://doi.org/10.1007/s00216-013-7271-9

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  • DOI: https://doi.org/10.1007/s00216-013-7271-9

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