Development of Multiwell-Plate Methods Using Pure Cultures of Methanogens To Identify New Inhibitors for Suppressing Ruminant Methane Emissions

Applied and Environmental Microbiology
M R WeimarGregory M Cook

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 librari...Continue Reading

References

Jan 1, 1986·Applied and Environmental Microbiology·T L MillerM P Bryant
Jul 31, 1980·Biochemical and Biophysical Research Communications·F D SauerJ D Erfle
Jun 6, 2000·Journal of Biomolecular Screening·J H ZhangK R Oldenburg
Sep 1, 2004·FEMS Microbiology Letters·Andrew K HaydockJohn A Leigh
Aug 1, 1990·Applied and Environmental Microbiology·K N JoblinA G Williams
Apr 22, 2006·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·F Sherwood Rowland
Apr 22, 2008·Applied and Environmental Microbiology·Peter H Janssen, Marek Kirs
Mar 30, 2010·The Veterinary Journal·Bryce M BuddleD Neil Wedlock
Mar 2, 2011·Nature Reviews. Drug Discovery·Ricardo MacarronG Sitta Sittampalam
Mar 16, 2011·Methods in Enzymology·Felipe B SarmientoWilliam B Whitman
Sep 29, 2011·The Journal of Biological Chemistry·Duncan G G McMillanGregory M Cook
May 16, 2012·Antimicrobial Agents and Chemotherapy·C BangR A Schmitz
Mar 15, 2013·Proceedings of the National Academy of Sciences of the United States of America·Felipe SarmientoWilliam B Whitman
Jun 14, 2013·Animal : an International Journal of Animal Bioscience·S C LeahyG T Attwood
Jul 16, 2013·Drug Discovery Today·Wei ZhengJohn C McKew
Aug 31, 2013·Standards in Genomic Sciences·S C LeahyG T Attwood
Nov 28, 2013·Animal : an International Journal of Animal Bioscience·J JeyanathanD P Morgavi
May 8, 2014·Archaea : an International Microbiological Journal·Leticia AbeciaDavid R Yáñez-Ruiz
Aug 1, 2015·Proceedings of the National Academy of Sciences of the United States of America·Alexander N HristovStephane Duval

❮ Previous
Next ❯

Citations

Jan 20, 2018·Archaea : an International Microbiological Journal·Feng LongWilliam B Whitman
Jun 28, 2019·The ISME Journal·Chris GreeningRoderick I Mackie
Oct 22, 2019·Frontiers in Microbiology·Natasha DoyleSinead Leahy
Nov 14, 2019·Animal Microbiome·William J KellyGraeme T Attwood
Mar 16, 2019·Current Opinion in Biotechnology·Zhe Lyu, William B Whitman

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.