Analysis of intracellular short organic acid-coenzyme a esters from actinomycetes using liquid chromatography-electrospray ionization-mass spectrometry

Won Park Je, Seok Jung Won, Ryeol Park Sung, Chul Park Byoung, Joon Yoon Yeo

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

A method employing silicone oil density centrifugation, solid-phase extraction (SPE) cleanup, and LC-ESI-MS/MS analysis was developed for the rapid, selective, sensitive, and quantitative detection of an intracellular pool of short organic acid-CoA esters in actinomycetes. The detection limit was determined to be approximately 0.8 pmol (1.2 ng/ml) for each standard CoA-ester analyzed by the present LC-ESI-MS/MS method. A selected ion chromatogram for a typical fragment ion (m/z 428) specific to CoA-esters enabled the detection of eight intracellular CoA-esters involved in both primary and secondary metabolisms. The application of this method to bacterial metabolomic study is demonstrated by the profiling of the intracellular CoA-ester pools in the wild-type Streptomyces venezuelae strain producing polyketide antibiotics (methymycin and pikromycin), a polyketide synthase (PKS)-deleted S. venezuelae mutant, and a S. venezuelae mutant expressing the heterologous PKS genes. By quantifying the individual CoA-esterlevel in three different genotypes of the S. Venezuela e strain, further insight could be gained into the role of CoA-estersin polyketide biosynthesis. This analytical approach can be extended to the quantification of the size and composition of in vivo CoA-ester pools in various microbes, and can provide a detailed understanding of the relationship between the in vivo CoA-ester pool and the production of pharmaceutically important polyketides.

Original languageEnglish
Pages (from-to)1136-1147
Number of pages12
JournalJournal of Mass Spectrometry
Volume42
Issue number9
DOIs
Publication statusPublished - 2007 Sep

Keywords

  • Actinomycetes
  • CoA-esters
  • LC-ESI-MS/MS
  • Metabolomics
  • Polyketide

ASJC Scopus subject areas

  • Spectroscopy

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