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Title: Dereplicating and spatial mapping of secondary metabolites from fungal cultures in situ

Abstract

Recently, ambient ionization mass spectrometry techniques have become prevalent in natural product research due to their ability to examine secondary metabolites in situ. Moreover, these techniques retain invaluable spatial and temporal details that are lost through traditional extraction processes. However, most ambient ionization techniques do not collect mutually supportive data, such as chromatographic retention times and/or UV/vis spectra, and this can limit the ability to identify certain metabolites, such as differentiating isomers. To overcome this, the droplet liquid microjunction-surface sampling probe (droplet-LMJ-SSP) was coupled with UPLC-PDA-HRMS-MS/MS, thus providing separation, retention times, MS data, and UV/vis data used in traditional dereplication protocols. By capturing these mutually supportive data, the identity of secondary metabolites can be confidently and rapidly assigned in situ. Using the droplet-LMJ-SSP, a protocol was constructed to analyze the secondary metabolite profile of fungal cultures without any sample preparation. Finally our results demonstrate that fungal cultures can be dereplicated from the Petri dish, thus identifying secondary metabolites, including isomers, and confirming them against reference standards. Furthermore, heat maps, similar to mass spectrometry imaging, can be used to ascertain the location and relative concentration of secondary metabolites directly on the surface and/or surroundings of a fungal culture.

Authors:
 [1];  [1];  [2];  [2];  [2];  [2];  [3]
  1. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  2. Univ. of North Carolina at Greensboro, Greensboro, NC (United States)
  3. Mycosynthetix, Inc., Hillsborough, NC (United States)
Publication Date:
Research Org.:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
Work for Others (WFO)
OSTI Identifier:
1214004
Alternate Identifier(s):
OSTI ID: 1287014
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Natural Products
Additional Journal Information:
Journal Volume: 78; Journal Issue: 8; Journal ID: ISSN 0163-3864
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; mass spectrometry; surface sampling; fungi; natural products; chemical imaging; 59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Kertesz, Vilmos, Van Berkel, Gary J., Sica, Vincent P., Raja, Huzefa A., El-Elimat, Tamam, Oberlies, Nicholas H., and Pearce, Cedric J. Dereplicating and spatial mapping of secondary metabolites from fungal cultures in situ. United States: N. p., 2015. Web. doi:10.1021/acs.jnatprod.5b00268.
Kertesz, Vilmos, Van Berkel, Gary J., Sica, Vincent P., Raja, Huzefa A., El-Elimat, Tamam, Oberlies, Nicholas H., & Pearce, Cedric J. Dereplicating and spatial mapping of secondary metabolites from fungal cultures in situ. United States. https://doi.org/10.1021/acs.jnatprod.5b00268
Kertesz, Vilmos, Van Berkel, Gary J., Sica, Vincent P., Raja, Huzefa A., El-Elimat, Tamam, Oberlies, Nicholas H., and Pearce, Cedric J. Thu . "Dereplicating and spatial mapping of secondary metabolites from fungal cultures in situ". United States. https://doi.org/10.1021/acs.jnatprod.5b00268. https://www.osti.gov/servlets/purl/1214004.
@article{osti_1214004,
title = {Dereplicating and spatial mapping of secondary metabolites from fungal cultures in situ},
author = {Kertesz, Vilmos and Van Berkel, Gary J. and Sica, Vincent P. and Raja, Huzefa A. and El-Elimat, Tamam and Oberlies, Nicholas H. and Pearce, Cedric J.},
abstractNote = {Recently, ambient ionization mass spectrometry techniques have become prevalent in natural product research due to their ability to examine secondary metabolites in situ. Moreover, these techniques retain invaluable spatial and temporal details that are lost through traditional extraction processes. However, most ambient ionization techniques do not collect mutually supportive data, such as chromatographic retention times and/or UV/vis spectra, and this can limit the ability to identify certain metabolites, such as differentiating isomers. To overcome this, the droplet liquid microjunction-surface sampling probe (droplet-LMJ-SSP) was coupled with UPLC-PDA-HRMS-MS/MS, thus providing separation, retention times, MS data, and UV/vis data used in traditional dereplication protocols. By capturing these mutually supportive data, the identity of secondary metabolites can be confidently and rapidly assigned in situ. Using the droplet-LMJ-SSP, a protocol was constructed to analyze the secondary metabolite profile of fungal cultures without any sample preparation. Finally our results demonstrate that fungal cultures can be dereplicated from the Petri dish, thus identifying secondary metabolites, including isomers, and confirming them against reference standards. Furthermore, heat maps, similar to mass spectrometry imaging, can be used to ascertain the location and relative concentration of secondary metabolites directly on the surface and/or surroundings of a fungal culture.},
doi = {10.1021/acs.jnatprod.5b00268},
journal = {Journal of Natural Products},
number = 8,
volume = 78,
place = {United States},
year = {Thu Jul 30 00:00:00 EDT 2015},
month = {Thu Jul 30 00:00:00 EDT 2015}
}

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Dereplication of macrocyclic trichothecenes from extracts of filamentous fungi through UV and NMR profiles
journal, July 2010

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Mass Spectrometry Sampling Under Ambient Conditions with Desorption Electrospray Ionization
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Ambient Mass Spectrometry
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Evaluation of culture media for the production of secondary metabolites in a natural products screening program
journal, January 2013

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Hydrophobins—Unique Fungal Proteins
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Burning the Hay to Find the Needle – Data Mining Strategies in Natural Product Dereplication
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The importance of mass spectrometric dereplication in fungal secondary metabolite analysis
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