Differential Off-line LC-NMR (DOLC-NMR) Metabolomics to Monitor Tyrosine-Induced Metabolome Alterations in Saccharomyces cerevisiae

Richard Hammerl, Oliver Frank, Thomas Hofmann

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

A novel differential off-line LC-NMR approach (DOLC-NMR) was developed to capture and quantify nutrient-induced metabolome alterations in Saccharomyces cerevisiae. Off-line coupling of HPLC separation and 1H NMR spectroscopy supported by automated comparative bucket analyses, followed by quantitative 1H NMR using ERETIC 2 (electronic reference to access in vivo concentrations), has been successfully used to quantitatively record changes in the metabolome of S. cerevisiae upon intervention with the aromatic amino acid l-tyrosine. Among the 33 metabolites identified, glyceryl succinate, tyrosol acetate, tyrosol lactate, tyrosol succinate, and N-acyl-tyrosine derivatives such as N-(1-oxooctyl)-tyrosine are reported for the first time as yeast metabolites. Depending on the chain length, N-(1-oxooctyl)-, N-(1-oxodecanyl)-, N-(1-oxododecanyl)-, N-(1-oxomyristinyl)-, N-(1-oxopalmityl)-, and N-(1-oxooleoyl)-l-tyrosine imparted a kokumi taste enhancement above their recognition thresholds ranging between 145 and 1432 μmol/L (model broth). Finally, carbon module labeling (CAMOLA) and carbon bond labeling (CABOLA) experiments with 13C6-glucose as the carbon source confirmed the biosynthetic pathway leading to the key metabolites; for example, the aliphatic side chain of N-(1-oxooctyl)-tyrosine could be shown to be generated via de novo fatty acid biosynthesis from four C2-carbon modules (acetyl-CoA) originating from glucose.

Original languageEnglish
Pages (from-to)3230-3241
Number of pages12
JournalJournal of agricultural and food chemistry
Volume65
Issue number15
DOIs
StatePublished - 19 Apr 2017

Keywords

  • C-labeling
  • CABOLA
  • CAMOLA
  • DOLC-NMR
  • Saccharomyces cerevisiae
  • kokumi
  • qNMR
  • quantitative NMR
  • taste
  • yeast

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