Abstract
Microbial cultures produce complex and potentially interesting mixtures of biosynthetic intermediates and derivatives of metabolites. These mixtures' reliable identification is important and so too is the development of techniques for their analysis. Here, a simple and highly selective method of detecting the biosynthetic congeners involved in the pentangular polyphenol pradimicin (PR) pathway from Actinomadura hibisca fermentation was developed. Solid-phase extraction (SPE) cleanup using an OASIS HLB cartridge was a simple and reliable tool for the extraction of PRs from a fermentation broth. The separation of each natural PR analog - eluted with a gradient system of aqueous acetonitrile through a reversed-phase C18 column containing ammonium acetate and acetic acid as additives - allowed their simultaneous profiling. The combined use of SPE cleanup and chromatographic separation, coupled with electrospray ionization-tandem mass spectrometry (ESI-MS/MS) detection was demonstrated to be sufficiently accurate and reliable to analyze the natural PR analogs produced from A. hibisca. Ten natural PRs were identified: four alanine-containing (PRA, PRC, PRL, and PRB), two glycine-substituted (PRD and PRE), and four serine-substituted (PRFA-1, PRFA-2, PRFL, and PRFB). This report demonstrates the first use of both SPE cleanup and HPLC-ESI-MS/MS to profile a wide range of structurally closely related PRs in a bacterial fermentation broth.
| Original language | English |
|---|---|
| Pages (from-to) | 2284-2291 |
| Number of pages | 8 |
| Journal | Journal of Chromatography A |
| Volume | 1218 |
| Issue number | 16 |
| DOIs | |
| Publication status | Published - 2011 Apr 22 |
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Keywords
- Actinomadura hibisca
- HPLC-ESI-MS/MS
- Metabolite profiling
- Pradimicins
ASJC Scopus subject areas
- Analytical Chemistry
- Organic Chemistry
- Biochemistry
Cite this
Characterization and identification of pradimicin analogs from Actinomadura hibisca using liquid chromatography-tandem mass spectrometry. / Park, Je Won; Park, Sung Ryeol; Han, Ah Reum; Ban, Yeon Hee; Yoo, Young Ji; Kim, Eunji; Kim, Beom Seok; Sohng, Jae Kyung; Yoon, Yeo Joon.
In: Journal of Chromatography A, Vol. 1218, No. 16, 22.04.2011, p. 2284-2291.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Characterization and identification of pradimicin analogs from Actinomadura hibisca using liquid chromatography-tandem mass spectrometry
AU - Park, Je Won
AU - Park, Sung Ryeol
AU - Han, Ah Reum
AU - Ban, Yeon Hee
AU - Yoo, Young Ji
AU - Kim, Eunji
AU - Kim, Beom Seok
AU - Sohng, Jae Kyung
AU - Yoon, Yeo Joon
PY - 2011/4/22
Y1 - 2011/4/22
N2 - Microbial cultures produce complex and potentially interesting mixtures of biosynthetic intermediates and derivatives of metabolites. These mixtures' reliable identification is important and so too is the development of techniques for their analysis. Here, a simple and highly selective method of detecting the biosynthetic congeners involved in the pentangular polyphenol pradimicin (PR) pathway from Actinomadura hibisca fermentation was developed. Solid-phase extraction (SPE) cleanup using an OASIS HLB cartridge was a simple and reliable tool for the extraction of PRs from a fermentation broth. The separation of each natural PR analog - eluted with a gradient system of aqueous acetonitrile through a reversed-phase C18 column containing ammonium acetate and acetic acid as additives - allowed their simultaneous profiling. The combined use of SPE cleanup and chromatographic separation, coupled with electrospray ionization-tandem mass spectrometry (ESI-MS/MS) detection was demonstrated to be sufficiently accurate and reliable to analyze the natural PR analogs produced from A. hibisca. Ten natural PRs were identified: four alanine-containing (PRA, PRC, PRL, and PRB), two glycine-substituted (PRD and PRE), and four serine-substituted (PRFA-1, PRFA-2, PRFL, and PRFB). This report demonstrates the first use of both SPE cleanup and HPLC-ESI-MS/MS to profile a wide range of structurally closely related PRs in a bacterial fermentation broth.
AB - Microbial cultures produce complex and potentially interesting mixtures of biosynthetic intermediates and derivatives of metabolites. These mixtures' reliable identification is important and so too is the development of techniques for their analysis. Here, a simple and highly selective method of detecting the biosynthetic congeners involved in the pentangular polyphenol pradimicin (PR) pathway from Actinomadura hibisca fermentation was developed. Solid-phase extraction (SPE) cleanup using an OASIS HLB cartridge was a simple and reliable tool for the extraction of PRs from a fermentation broth. The separation of each natural PR analog - eluted with a gradient system of aqueous acetonitrile through a reversed-phase C18 column containing ammonium acetate and acetic acid as additives - allowed their simultaneous profiling. The combined use of SPE cleanup and chromatographic separation, coupled with electrospray ionization-tandem mass spectrometry (ESI-MS/MS) detection was demonstrated to be sufficiently accurate and reliable to analyze the natural PR analogs produced from A. hibisca. Ten natural PRs were identified: four alanine-containing (PRA, PRC, PRL, and PRB), two glycine-substituted (PRD and PRE), and four serine-substituted (PRFA-1, PRFA-2, PRFL, and PRFB). This report demonstrates the first use of both SPE cleanup and HPLC-ESI-MS/MS to profile a wide range of structurally closely related PRs in a bacterial fermentation broth.
KW - Actinomadura hibisca
KW - HPLC-ESI-MS/MS
KW - Metabolite profiling
KW - Pradimicins
UR - http://www.scopus.com/inward/record.url?scp=79953077645&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79953077645&partnerID=8YFLogxK
U2 - 10.1016/j.chroma.2011.02.034
DO - 10.1016/j.chroma.2011.02.034
M3 - Article
C2 - 21376331
AN - SCOPUS:79953077645
VL - 1218
SP - 2284
EP - 2291
JO - Journal of Chromatography
JF - Journal of Chromatography
SN - 0021-9673
IS - 16
ER -