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:
-
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Univ. of North Carolina at Greensboro, Greensboro, NC (United States)
- 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}
}
Web of Science
Works referenced in this record:
On-line screening for drug metabolites by high-performance liquid chromatography with a diode array UV detector
journal, December 1983
- Zech, Karl; Huber, Reinhard; Elgass, H.
- Journal of Chromatography A, Vol. 282
Burning the Hay to Find the Needle – Data Mining Strategies in Natural Product Dereplication
journal, June 2007
- Wolf, D.; Siems, K.
- CHIMIA International Journal for Chemistry, Vol. 61, Issue 6
Hyphenated NMR Methods in Natural Products Research, Part 1: Direct Hyphenation
journal, July 2005
- Jaroszewski, Jerzy W.
- Planta Medica, Vol. 71, Issue 8
Evolving Trends in the Dereplication of Natural Product Extracts: New Methodology for Rapid, Small-Scale Investigation of Natural Product Extracts
journal, August 2008
- Lang, Gerhard; Mayhudin, Nor Ainy; Mitova, Maya I.
- Journal of Natural Products, Vol. 71, Issue 9
High-Resolution MS, MS/MS, and UV Database of Fungal Secondary Metabolites as a Dereplication Protocol for Bioactive Natural Products
journal, August 2013
- El-Elimat, Tamam; Figueroa, Mario; Ehrmann, Brandie M.
- Journal of Natural Products, Vol. 76, Issue 9
Dereplication of macrocyclic trichothecenes from extracts of filamentous fungi through UV and NMR profiles
journal, July 2010
- Sy-Cordero, Arlene A.; Graf, Tyler N.; Wani, Mansukh C.
- The Journal of Antibiotics, Vol. 63, Issue 9
Emerging capabilities of mass spectrometry for natural products
journal, January 2014
- Jarmusch, Alan K.; Cooks, R. Graham
- Nat. Prod. Rep., Vol. 31, Issue 6
Direct mass spectrometric screening of antibiotics from bacterial surfaces using liquid extraction surface analysis: Screening of antibiotics using LESA
journal, September 2012
- Kai, Marco; González, Ignacio; Genilloud, Olga
- Rapid Communications in Mass Spectrometry, Vol. 26, Issue 20
Real-Time Metabolomics on Living Microorganisms Using Ambient Electrospray Ionization Flow-Probe
journal, July 2013
- Hsu, Cheng-Chih; ElNaggar, Mariam S.; Peng, Yao
- Analytical Chemistry, Vol. 85, Issue 15
Molecular Networking as a Dereplication Strategy
journal, August 2013
- Yang, Jane Y.; Sanchez, Laura M.; Rath, Christopher M.
- Journal of Natural Products, Vol. 76, Issue 9
Mass spectrometry imaging of secondary metabolites directly on fungal cultures
journal, January 2014
- Sica, V. P.; Raja, H. A.; El-Elimat, T.
- RSC Adv., Vol. 4, Issue 108
Visualizing life with ambient mass spectrometry
journal, February 2015
- Hsu, Cheng-Chih; Dorrestein, Pieter C.
- Current Opinion in Biotechnology, Vol. 31
Desorption electrospray ionization mass spectrometry: Imaging drugs and metabolites in tissues
journal, August 2008
- Wiseman, J. M.; Ifa, D. R.; Zhu, Y.
- Proceedings of the National Academy of Sciences, Vol. 105, Issue 47
Polyhydroxyanthraquinones as Quorum Sensing Inhibitors from the Guttates of Penicillium restrictum and Their Analysis by Desorption Electrospray Ionization Mass Spectrometry
journal, June 2014
- Figueroa, Mario; Jarmusch, Alan K.; Raja, Huzefa A.
- Journal of Natural Products, Vol. 77, Issue 6
The importance of mass spectrometric dereplication in fungal secondary metabolite analysis
journal, February 2015
- Nielsen, Kristian F.; Larsen, Thomas O.
- Frontiers in Microbiology, Vol. 6
Liquid Microjunction Surface Sampling Coupled with High-Pressure Liquid Chromatography−Electrospray Ionization-Mass Spectrometry for Analysis of Drugs and Metabolites in Whole-Body Thin Tissue Sections
journal, July 2010
- Kertesz, Vilmos; Van Berkel, Gary J.
- Analytical Chemistry, Vol. 82, Issue 14
Automated liquid microjunction surface sampling-HPLC–MS/MS analysis of drugs and metabolites in whole-body thin tissue sections
journal, April 2013
- Kertesz, Vilmos; Van Berkel, Gary J.
- Bioanalysis, Vol. 5, Issue 7
Sampling reliability, spatial resolution, spatial precision, and extraction efficiency in droplet-based liquid microjunction surface sampling: Basics of LMJ/HPLC/MS/MS sampling
journal, May 2014
- Kertesz, Vilmos; Van Berkel, Gary J.
- Rapid Communications in Mass Spectrometry, Vol. 28, Issue 13
Liquid microjunction surface sampling of acetaminophen, terfenadine and their metabolites in thin tissue sections
journal, October 2014
- Kertesz, Vilmos; Paranthaman, Nithya; Moench, Paul
- Bioanalysis, Vol. 6, Issue 19
An enhanced droplet-based liquid microjunction surface sampling system coupled with HPLC-ESI-MS/MS for spatially resolved analysis
journal, November 2014
- Kertesz, Vilmos; Weiskittel, Taylor M.; Van Berkel, Gary J.
- Analytical and Bioanalytical Chemistry, Vol. 407, Issue 8
Mass Spectrometry Sampling Under Ambient Conditions with Desorption Electrospray Ionization
journal, October 2004
- Takats, Z.
- Science, Vol. 306, Issue 5695, p. 471-473
Nanospray desorption electrospray ionization: an ambient method for liquid-extraction surface sampling in mass spectrometry
journal, January 2010
- Roach, Patrick J.; Laskin, Julia; Laskin, Alexander
- The Analyst, Vol. 135, Issue 9
MALDI imaging mass spectrometry: molecular snapshots of biochemical systems
journal, September 2007
- Cornett, Dale S.; Reyzer, Michelle L.; Chaurand, Pierre
- Nature Methods, Vol. 4, Issue 10
Laser Ablation Electrospray Ionization for Atmospheric Pressure, in Vivo, and Imaging Mass Spectrometry
journal, November 2007
- Nemes, Peter; Vertes, Akos
- Analytical Chemistry, Vol. 79, Issue 21
Fully automated liquid extraction-based surface sampling and ionization using a chip-based robotic nanoelectrospray platform
journal, March 2010
- Kertesz, Vilmos; Van Berkel, Gary J.
- Journal of Mass Spectrometry, Vol. 45, Issue 3
Imaging Mass Spectrometry of a Coral Microbe Interaction with Fungi
journal, July 2013
- Moree, Wilna J.; Yang, Jane Y.; Zhao, Xiling
- Journal of Chemical Ecology, Vol. 39, Issue 7
Chemical Diversity of Metabolites from Fungi, Cyanobacteria, and Plants Relative to FDA-Approved Anticancer Agents
journal, July 2012
- El-Elimat, Tamam; Zhang, Xiaoli; Jarjoura, David
- ACS Medicinal Chemistry Letters, Vol. 3, Issue 8
Greensporones: Resorcylic Acid Lactones from an Aquatic Halenospora sp.
journal, August 2014
- El-Elimat, Tamam; Raja, Huzefa A.; Day, Cynthia S.
- Journal of Natural Products, Vol. 77, Issue 9
New diketopiperazine dimer from a filamentous fungal isolate of Aspergillus sydowii : Diketopiperazine dimer
journal, June 2015
- Kaur, Amninder; Raja, Huzefa A.; Darveaux, Blaise A.
- Magnetic Resonance in Chemistry, Vol. 53, Issue 8
Flavonolignans from Aspergillus iizukae , a Fungal Endophyte of Milk Thistle ( Silybum marianum )
journal, January 2014
- El-Elimat, Tamam; Raja, Huzefa A.; Graf, Tyler N.
- Journal of Natural Products, Vol. 77, Issue 2
Evaluation of culture media for the production of secondary metabolites in a natural products screening program
journal, January 2013
- VanderMolen, Karen M.; Raja, Huzefa A.; El-Elimat, Tamam
- AMB Express, Vol. 3, Issue 1
Cytotoxic epipolythiodioxopiperazine alkaloids from filamentous fungi of the Bionectriaceae
journal, September 2012
- Figueroa, Mario; Graf, Tyler N.; Ayers, Sloan
- The Journal of Antibiotics, Vol. 65, Issue 11
Peptaibols from two unidentified fungi of the order Hypocreales with cytotoxic, antibiotic, and anthelmintic activities: PEPTAIBOLS WITH CYTOTOXIC, ANTIBIOTIC, AND ANTHELMINTIC ACTIVITIES
journal, June 2012
- Ayers, Sloan; Ehrmann, Brandie M.; Adcock, Audrey F.
- Journal of Peptide Science, Vol. 18, Issue 8
Mass spectral molecular networking of living microbial colonies
journal, May 2012
- Watrous, J.; Roach, P.; Alexandrov, T.
- Proceedings of the National Academy of Sciences, Vol. 109, Issue 26
Size, weight and position: ion mobility spectrometry and imaging MS combined
journal, January 2011
- Kiss, András; Heeren, Ron M. A.
- Analytical and Bioanalytical Chemistry, Vol. 399, Issue 8
Mass spectrometry-based proteomics: existing capabilities and future directions
journal, January 2012
- Angel, Thomas E.; Aryal, Uma K.; Hengel, Shawna M.
- Chemical Society Reviews, Vol. 41, Issue 10
Hydrophobins—Unique Fungal Proteins
journal, May 2012
- Bayry, Jagadeesh; Aimanianda, Vishukumar; Guijarro, J. Iñaki
- PLoS Pathogens, Vol. 8, Issue 5
Phenotypic taxonomy and metabolite profiling in microbial drug discovery
journal, January 2005
- Larsen, Thomas O.; Smedsgaard, Jørn; Nielsen, Kristian F.
- Natural Product Reports, Vol. 22, Issue 6
Phylogenetic and chemical diversity of fungal endophytes isolated from Silybum marianum (L) Gaertn. (milk thistle)
journal, January 2015
- Raja, Huzefa A.; Kaur, Amninder; El-Elimat, Tamam
- Mycology, Vol. 6, Issue 1
Phenotypic Taxonomy and Metabolite Profiling in Microbial Drug Discovery
journal, April 2006
- Larsen, Thomas O.; Smedsgaard, Joern; Nielsen, Kristian F.
- ChemInform, Vol. 37, Issue 14
Fully automated liquid extraction-based surface sampling and ionization using a chip-based robotic nanoelectrospray platform
journal, March 2010
- Kertesz, Vilmos; Van Berkel, Gary J.
- Journal of Mass Spectrometry, Vol. 45, Issue 3
New diketopiperazine dimer from a filamentous fungal isolate of Aspergillus sydowii : Diketopiperazine dimer
journal, June 2015
- Kaur, Amninder; Raja, Huzefa A.; Darveaux, Blaise A.
- Magnetic Resonance in Chemistry, Vol. 53, Issue 8
Peptaibols from two unidentified fungi of the order Hypocreales with cytotoxic, antibiotic, and anthelmintic activities: PEPTAIBOLS WITH CYTOTOXIC, ANTIBIOTIC, AND ANTHELMINTIC ACTIVITIES
journal, June 2012
- Ayers, Sloan; Ehrmann, Brandie M.; Adcock, Audrey F.
- Journal of Peptide Science, Vol. 18, Issue 8
An enhanced droplet-based liquid microjunction surface sampling system coupled with HPLC-ESI-MS/MS for spatially resolved analysis
journal, November 2014
- Kertesz, Vilmos; Weiskittel, Taylor M.; Van Berkel, Gary J.
- Analytical and Bioanalytical Chemistry, Vol. 407, Issue 8
Fungal metabolite screening: database of 474 mycotoxins and fungal metabolites for dereplication by standardised liquid chromatography–UV–mass spectrometry methodology
journal, June 2003
- Nielsen, K.
- Journal of Chromatography A, Vol. 1002, Issue 1-2
Micro-scale extraction procedure for standardized screening of fungal metabolite production in cultures
journal, January 1997
- Smedsgaard, Jørn
- Journal of Chromatography A, Vol. 760, Issue 2
Laser Ablation Electrospray Ionization for Atmospheric Pressure, in Vivo, and Imaging Mass Spectrometry
journal, November 2007
- Nemes, Peter; Vertes, Akos
- Analytical Chemistry, Vol. 79, Issue 21
Real-Time Metabolomics on Living Microorganisms Using Ambient Electrospray Ionization Flow-Probe
journal, July 2013
- Hsu, Cheng-Chih; ElNaggar, Mariam S.; Peng, Yao
- Analytical Chemistry, Vol. 85, Issue 15
Molecular Networking as a Dereplication Strategy
journal, August 2013
- Yang, Jane Y.; Sanchez, Laura M.; Rath, Christopher M.
- Journal of Natural Products, Vol. 76, Issue 9
Flavonolignans from Aspergillus iizukae , a Fungal Endophyte of Milk Thistle ( Silybum marianum )
journal, January 2014
- El-Elimat, Tamam; Raja, Huzefa A.; Graf, Tyler N.
- Journal of Natural Products, Vol. 77, Issue 2
Polyhydroxyanthraquinones as Quorum Sensing Inhibitors from the Guttates of Penicillium restrictum and Their Analysis by Desorption Electrospray Ionization Mass Spectrometry
journal, June 2014
- Figueroa, Mario; Jarmusch, Alan K.; Raja, Huzefa A.
- Journal of Natural Products, Vol. 77, Issue 6
Greensporones: Resorcylic Acid Lactones from an Aquatic Halenospora sp.
journal, August 2014
- El-Elimat, Tamam; Raja, Huzefa A.; Day, Cynthia S.
- Journal of Natural Products, Vol. 77, Issue 9
Dereplication of macrocyclic trichothecenes from extracts of filamentous fungi through UV and NMR profiles
journal, July 2010
- Sy-Cordero, Arlene A.; Graf, Tyler N.; Wani, Mansukh C.
- The Journal of Antibiotics, Vol. 63, Issue 9
MALDI imaging mass spectrometry: molecular snapshots of biochemical systems
journal, September 2007
- Cornett, Dale S.; Reyzer, Michelle L.; Chaurand, Pierre
- Nature Methods, Vol. 4, Issue 10
Nanospray desorption electrospray ionization: an ambient method for liquid-extraction surface sampling in mass spectrometry
journal, January 2010
- Roach, Patrick J.; Laskin, Julia; Laskin, Alexander
- The Analyst, Vol. 135, Issue 9
Emerging capabilities of mass spectrometry for natural products
journal, January 2014
- Jarmusch, Alan K.; Cooks, R. Graham
- Nat. Prod. Rep., Vol. 31, Issue 6
Hyphenated NMR Methods in Natural Products Research, Part 1: Direct Hyphenation
journal, July 2005
- Jaroszewski, Jerzy W.
- Planta Medica, Vol. 71, Issue 8
Mass spectral molecular networking of living microbial colonies
journal, May 2012
- Watrous, J.; Roach, P.; Alexandrov, T.
- Proceedings of the National Academy of Sciences, Vol. 109, Issue 26
Phylogenetic and chemical diversity of fungal endophytes isolated from Silybum marianum (L) Gaertn. (milk thistle)
journal, January 2015
- Raja, Huzefa A.; Kaur, Amninder; El-Elimat, Tamam
- Mycology, Vol. 6, Issue 1
Mass Spectrometry Sampling Under Ambient Conditions with Desorption Electrospray Ionization
journal, October 2004
- Takats, Z.
- Science, Vol. 306, Issue 5695, p. 471-473
Evaluation of culture media for the production of secondary metabolites in a natural products screening program
journal, January 2013
- VanderMolen, Karen M.; Raja, Huzefa A.; El-Elimat, Tamam
- AMB Express, Vol. 3, Issue 1
Hydrophobins—Unique Fungal Proteins
journal, May 2012
- Bayry, Jagadeesh; Aimanianda, Vishukumar; Guijarro, J. Iñaki
- PLoS Pathogens, Vol. 8, Issue 5
Burning the Hay to Find the Needle – Data Mining Strategies in Natural Product Dereplication
journal, June 2007
- Wolf, D.; Siems, K.
- CHIMIA International Journal for Chemistry, Vol. 61, Issue 6
The importance of mass spectrometric dereplication in fungal secondary metabolite analysis
journal, February 2015
- Nielsen, Kristian F.; Larsen, Thomas O.
- Frontiers in Microbiology, Vol. 6
Works referencing / citing this record:
Review and perspectives on the applications of mass spectrometry imaging under ambient conditions
journal, July 2018
- Perez, Consuelo J.; Bagga, Aafreen K.; Prova, Shamina S.
- Rapid Communications in Mass Spectrometry, Vol. 33, Issue S3
Spatial and Temporal Profiling of Griseofulvin Production in Xylaria cubensis Using Mass Spectrometry Mapping
journal, April 2016
- Sica, Vincent P.; Rees, Evan R.; Tchegnon, Edem
- Frontiers in Microbiology, Vol. 7
Mapping the Fungal Battlefield: Using in situ Chemistry and Deletion Mutants to Monitor Interspecific Chemical Interactions Between Fungi
journal, February 2019
- Knowles, Sonja L.; Raja, Huzefa A.; Wright, Allison J.
- Frontiers in Microbiology, Vol. 10
Droplet probe: coupling chromatography to the in situ evaluation of the chemistry of nature
journal, January 2019
- Oberlies, Nicholas H.; Knowles, Sonja L.; Amrine, Chiraz Soumia M.
- Natural Product Reports, Vol. 36, Issue 7
Media studies to enhance the production of verticillins facilitated by in situ chemical analysis
journal, September 2018
- Amrine, Chiraz Soumia M.; Raja, Huzefa A.; Darveaux, Blaise A.
- Journal of Industrial Microbiology & Biotechnology, Vol. 45, Issue 12
Rapid Characterization of Chemical Components in Edible Mushroom Sparassis crispa by UPLC-Orbitrap MS Analysis and Potential Inhibitory Effects on Allergic Rhinitis
journal, August 2019
- Wang, Zhixin; Liu, Jingyu; Zhong, Xiangjian
- Molecules, Vol. 24, Issue 16
Media studies to enhance the production of verticillins facilitated by in situ chemical analysis
journal, September 2018
- Amrine, Chiraz Soumia M.; Raja, Huzefa A.; Darveaux, Blaise A.
- Journal of Industrial Microbiology & Biotechnology, Vol. 45, Issue 12
Biosynthesis of Fluorinated Peptaibols Using a Site-Directed Building Block Incorporation Approach
journal, June 2017
- Rivera-Chávez, José; Raja, Huzefa A.; Graf, Tyler N.
- Journal of Natural Products, Vol. 80, Issue 6
Enhanced dereplication of fungal cultures via use of mass defect filtering
journal, January 2017
- Paguigan, Noemi D.; El-Elimat, Tamam; Kao, Diana
- The Journal of Antibiotics, Vol. 70, Issue 5
Spatial and Temporal Profiling of Griseofulvin Production in Xylaria cubensis Using Mass Spectrometry Mapping
journal, April 2016
- Sica, Vincent P.; Rees, Evan R.; Tchegnon, Edem
- Frontiers in Microbiology, Vol. 7
Xanthones of Lichen Source: A 2016 Update
journal, March 2016
- Le Pogam, Pierre; Boustie, Joël
- Molecules, Vol. 21, Issue 3