Simultaneous monitoring of the bioconversion from lysine to glutaric acid by ethyl chloroformate derivatization and gas chromatography-mass spectrometry

Yeong Hoon Han, Tae Rim Choi, Ye Lim Park, Hun Suk Song, Yong Keun Choi, Hyun Joong Kim, Shashi Kant Bhatia, Ranjit Gurav, Kyungmoon Park, See Hyoung Park, Wooseong Kim, Yung Hun Yang

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Glutaric acid is a precursor of a plasticizer that can be used for the production of polyester amides, ester plasticizer, corrosion inhibitor, and others. Glutaric acid can be produced either via bioconversion or chemical synthesis, and some metabolites and intermediates are produced during the reaction. To ensure reaction efficiency, the substrates, intermediates, and products, especially in the bioconversion system, should be closely monitored. Until now, high performance liquid chromatography (HPLC) has generally been used to analyze the glutaric acid-related metabolites, although it demands separate time-consuming derivatization and non-derivatization analyses. To substitute for this unreasonable analytical method, we applied herein a gas chromatography - mass spectrometry (GC-MS) method with ethyl chloroformate (ECF) derivatization to simultaneously monitor the major metabolites. We determined the suitability of GC-MS analysis using defined concentrations of six metabolites (L-lysine, cadaverine, 5-aminovaleric acid, 2-oxoglutaric acid, glutamate, and glutaric acid) and their mass chromatograms, regression equations, regression coefficient values (R2), dynamic ranges (mM), and retention times (RT). This method successfully monitored the production process in complex fermentation broth.

Original languageEnglish
Article number113688
JournalAnalytical Biochemistry
Volume597
DOIs
StatePublished - 15 May 2020

Keywords

  • Analytical method
  • Ethyl chloroformate
  • Gas chromatography-mass spectrometry
  • Glutaric acid
  • Mass chromatograms
  • Metabolites

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