Development of multiwell-plate methods using pure cultures of methanogens to identify new inhibitors for suppressing ruminant methane emissions

Weimar, M. R., Cheung, J., Dey, D., McSweeney, C., Morrison, M., Kobayashi, Y., Whitman, W. B., Carbone, V., Schofield, L. R., Ronimus, R. S. and Cook, G. M. (2017) Development of multiwell-plate methods using pure cultures of methanogens to identify new inhibitors for suppressing ruminant methane emissions. Applied and Environmental Microbiology, 83 15: 722-725. doi:10.1128/AEM.00396-17


Author Weimar, M. R.
Cheung, J.
Dey, D.
McSweeney, C.
Morrison, M.
Kobayashi, Y.
Whitman, W. B.
Carbone, V.
Schofield, L. R.
Ronimus, R. S.
Cook, G. M.
Title Development of multiwell-plate methods using pure cultures of methanogens to identify new inhibitors for suppressing ruminant methane emissions
Journal name Applied and Environmental Microbiology   Check publisher's open access policy
ISSN 1098-5336
Publication date 2017-08-01
Year available 2008
Sub-type Article (original research)
DOI 10.1128/AEM.00396-17
Open Access Status Not yet assessed
Volume 83
Issue 15
Start page 722
End page 725
Total pages 8
Place of publication Washington, DC, United States
Publisher American Society for Microbiology
Language eng
Subject 1305 Biotechnology
1106 Food Science
2402 Applied Microbiology and Biotechnology
2303 Ecology
Abstract Hydrogenotrophic methanogens typically require strictly anaerobic culturing conditions in glass tubes with overpressures of H and CO that are both time-consuming and costly. To increase the throughput for screening chemical compound libraries, 96-well microtiter plate methods for the growth of a marine (environmental) methanogen Methanococcus maripaludis strain S2 and the rumen methanogen Methanobrevibacter species AbM4 were developed. A number of key parameters (inoculum size, reducing agents for medium preparation, assay duration, inhibitor solvents, and culture volume) were optimized to achieve robust and reproducible growth in a high-throughput microtiter plate format. The method was validated using published methanogen inhibitors and statistically assessed for sensitivity and reproducibility. The Sigma-Aldrich LOPAC library containing 1,280 pharmacologically active compounds and an in-house natural product library (120 compounds) were screened against M. maripaludis as a proof of utility. This screen identified a number of bioactive compounds, and MIC values were confirmed for some of them against M. maripaludis and M. AbM4. The developed method provides a significant increase in throughput for screening compound libraries and can now be used to screen larger compound libraries to discover novel methanogen-specific inhibitors for the mitigation of ruminant methane emissions.
Formatted abstract
Hydrogenotrophic methanogens typically require strictly anaerobic culturing conditions in glass tubes with overpressures of H2 and CO2 that are both time-consuming and costly. To increase the throughput for screening chemical compound libraries, 96-well microtiter plate methods for the growth of a marine (environmental) methanogen Methanococcus maripaludis strain S2 and the rumen methanogen Methanobrevibacter species AbM4 were developed. A number of key parameters (inoculum size, reducing agents for medium preparation, assay duration, inhibitor solvents, and culture volume) were optimized to achieve robust and reproducible growth in a high-throughput microtiter plate format. The method was validated using published methanogen inhibitors and statistically assessed for sensitivity and reproducibility. The Sigma-Aldrich LOPAC library containing 1,280 pharmacologically active compounds and an in-house natural product library (120 compounds) were screened against M. maripaludis as a proof of utility. This screen identified a number of bioactive compounds, and MIC values were confirmed for some of them against M. maripaludis and M. AbM4. The developed method provides a significant increase in throughput for screening compound libraries and can now be used to screen larger compound libraries to discover novel methanogen-specific inhibitors for the mitigation of ruminant methane emissions.
Keyword Greenhouse gas
Highthroughput
Methanobrevibacter
Methanococcus maripaludis
Methanogen
Rumen
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ
Additional Notes Article number e00396-17

Document type: Journal Article
Sub-type: Article (original research)
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