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Appropriate sampling for intracellular amino acid analysis in five phylogenetically different yeasts

Authors
  • Bolten, Christoph J.1, 2
  • Wittmann, Christoph1
  • 1 Westfalian Wilhelms-University, Biotechnology Department, Biochemistry Institute, Munster, Germany , Munster (Germany)
  • 2 Saarland University, Biochemical Engineering, Saarbrucken, Germany , Saarbrucken (Germany)
Type
Published Article
Journal
Biotechnology Letters
Publisher
Springer-Verlag
Publication Date
Jul 05, 2008
Volume
30
Issue
11
Pages
1993–2000
Identifiers
DOI: 10.1007/s10529-008-9789-z
Source
Springer Nature
Keywords
License
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Abstract

Methanol quenching and fast filtration, the two most common sampling protocols in microbial metabolome analysis, were validated for intracellular amino acid analysis in phylogenetically different yeast strains comprising Saccharomyces cerevisiae, Kluyveromyces marxianus, Pichia pastoris, Schizosaccharomyces pombe and Zygosaccharomyces bailii. With only few exceptions for selected amino acids, all yeasts exhibited negligible metabolite leakage during quenching with 60% cold buffered methanol. Slightly higher leakage was observed with increasing methanol content in the quenching solution. Fast filtration resulted in identical levels for intracellular amino acids in all strains tested. The results clearly demonstrate the validity of both approaches for leakage-free sampling of amino acids in yeast.

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