Original Article

Metabolomics

, Volume 4, Issue 3, pp 240-247

Leakage of adenylates during cold methanol/glycerol quenching of Escherichia coli

  • Hannes LinkAffiliated withLehrstuhl für Bioverfahrenstechnik, Technische Universität München
  • , Bernd AnselmentAffiliated withLehrstuhl für Bioverfahrenstechnik, Technische Universität München
  • , Dirk Weuster-BotzAffiliated withLehrstuhl für Bioverfahrenstechnik, Technische Universität München Email author 

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Abstract

Effective and rapid inactivation of cellular metabolism is a prerequisite for accurate metabolome analysis. Cold methanol quenching is commonly applied to stop any metabolic activity and, at the same time remaining the cells’ integrity. However, it is reported that especially prokaryotic cells like Escherichia coli and Corynebacterium glutamicum tend to leak intracellular metabolites during cold methanol quenching. In this work leakage of adenylates is quantified for different quenching fluids. Further, a methanol/glycerol based quenching fluid is proposed, which reduces leakage drastically compared to the commonly applied methanol/water solution (16% ATP leakage compared to more than 70%).

Keywords

Quenching Cold shock Cell leakage Adenylates Glycerol