Liquid chromatography-mass spectrometry characterization of FK506 biosynthetic intermediates in Streptomyces clavuligerus KCTC 10561BP

Je Won Park, Sang Joon Mo, Sung Ryeol Park, Yeon Hee Ban, Young Ji Yoo, Yeo Joon Yoon

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The development of an efficient analytical method for the reliable detection and identification of the biosynthetic intermediates found in microbial cultures, which usually produce complex intermediates of the metabolites of interest, is essential for further biosynthetic investigations. This study developed a simple and highly selective method for detecting the biosynthetic intermediates involved in the FK506 pathway of Streptomyces clavuligerus KCTC 10561BP involving a cleanup procedure using a solid-phase extraction technique to provide reliable extraction of FK506-related compounds from a cell culture broth and liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) to separate and detect the FK506-related intermediates at concentrations as low as 0.2 μg/L in the broth. This method enabled the analytical profiling of the intermediates formed during the biosynthesis of FK506 in this S. clavuligerus strain, which produced FK506 as a main product. Eight FK506 intermediates-FK520, 37,38-dihydroFK506, prolylFK506, 9-decarbonyl-9-hydroxylFK506, 9-deoxoFK506, desmethylFK520, prolylFK520, and 9-deoxoFK520-were identified. This is the first report of the LC-ESI-MS/MS characterization of a wide range of FK506 analogs from a bacterial fermentation broth. The protocol employed in this study may be useful for estimating the structure of the metabolites without the need for a time-consuming isolation process and nuclear magnetic resonance (NMR) spectroscopy.

Original languageEnglish
Pages (from-to)1-7
Number of pages7
JournalAnalytical Biochemistry
Volume393
Issue number1
DOIs
Publication statusPublished - 2009 Oct 1
Externally publishedYes

Fingerprint

Liquid chromatography
Streptomyces
Tacrolimus
Liquid Chromatography
Mass spectrometry
Mass Spectrometry
Metabolites
Electrospray ionization
Electrospray Ionization Mass Spectrometry
Biosynthesis
Solid Phase Extraction
Tandem Mass Spectrometry
Cell culture
Fermentation
Nuclear magnetic resonance spectroscopy
Magnetic Resonance Spectroscopy
Cell Culture Techniques

Keywords

  • FK506 profiling
  • Liquid chromatography-mass spectrometry (LC-MS)
  • Tacrolimus

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

Cite this

Liquid chromatography-mass spectrometry characterization of FK506 biosynthetic intermediates in Streptomyces clavuligerus KCTC 10561BP. / Park, Je Won; Mo, Sang Joon; Park, Sung Ryeol; Ban, Yeon Hee; Yoo, Young Ji; Yoon, Yeo Joon.

In: Analytical Biochemistry, Vol. 393, No. 1, 01.10.2009, p. 1-7.

Research output: Contribution to journalArticle

Park, Je Won ; Mo, Sang Joon ; Park, Sung Ryeol ; Ban, Yeon Hee ; Yoo, Young Ji ; Yoon, Yeo Joon. / Liquid chromatography-mass spectrometry characterization of FK506 biosynthetic intermediates in Streptomyces clavuligerus KCTC 10561BP. In: Analytical Biochemistry. 2009 ; Vol. 393, No. 1. pp. 1-7.
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