Embedded Files
Research Team
Research Team
Hailey Hodulik
Hailey Hodulik
Biology
Debdeep Das
Debdeep Das
Microbiology
Abstract/Introduction
Abstract/Introduction
Methanogenesis contributes to greenhouse gas production
Methane production can occur through degradation of compounds such as acetate, methanol, & methlyamines
Proteomic studies of Methanococcoides methylutens Q3c showed upregulation of proteins MttB and MttC
Understand the enzymatic pathway of dimethylethanolamine (DMEA) degradation
Methods
Methods
Transformation and protein expression:
WWM73 transformed with plasmids containing 2119 & 2117
Cultures grown anaerobically, in high-salt conditions, and with puromycin selection
Protein purification:
Cells harvested & lysed with French Press
Purification with Ni-NTA affinity chromatography
Enzyme activity assays:
Ellman’s assay to detect methyl-coenzyme M
Product verification:
HPLC or NMR analysis will be used to verify DMEA → MMEA
Results/Findings and Discussion
Results/Findings and Discussion
Volcano plots show upregulated proteins when grown with choline or QMA. Upregulated (red) and downregulated (blue) in each growth condition.
Plot A: choline vs TMA
Plot B: QMA vs TMA
Conclusions and Future Study
Conclusions and Future Study
MttB and MttC are likely involved in DMEA metabolism
Successful transformation and partial purification
DMEA Catabolism
DMEA Catabolism
Image indicates the degradation of dimethylethanolamine (DMEA). The first step of this process is the demethylation of DMEA into monomethylethanolamine (MMEA) by methyltransferase 1 (MT1/2119/MttB). MT1 transfers the methyl group to a coronoid binding protein (2117 or MttC). MttC has an inner core with cobalt, which must be activated by a protein called Ram. Ram activates cobalt by transferring electrons from ferrodoxin. Then an enzyme called methyl transferase 2 transfers the methyl group to Coenzyme-M. Coenzyme-M and CH3 are reduced by MCR, which is a reductase. This produces methane gas as a byproduct.
The following is an image of poster presented at the 2026 Undergraduate Research Forum
Acknowledgements
Acknowledgements
Miami University, NSF, and CBFG
References
References
Buan NR. Methanogens: pushing the boundaries of biology. Emerg Top Life Sci. 2018 Dec 14;2(4):629-646. doi: 10.1042/ETLS20180031. PMID: 33525834; PMCID: PMC7289024.
Ferguson DJ Jr, Gorlatova N, Grahame DA, Krzycki JA. Reconstitution of dimethylamine:coenzyme M methyl transfer with a discrete corrinoid protein and two methyltransferases purified from Methanosarcina barkeri. J Biol Chem. 2000 Sep 15;275(37):29053-60. doi: 10.1074/jbc.M910218199. PMID: 10852929.
Chao, J. New study finds the economic harms of methane can vary greatly by region. 2021, May 21. https://climatesciences.lbl.gov/2021/05/21/new-study-finds-the-economic-harms-of-methane-emissions-can-vary-greatly-by-region/.
NACE Career Readiness Competencies
NACE Career Readiness Competencies
Communication: Throughout the course of this project I have developed my communication skills by communicating with those I am collaborating on the project with and by communicating the process and results of my research.
Teamwork: Working on this research project has strengthed my ability to work in a team, which is essential for any future career.
Critical Thinking: This research project has helped me engage in critical thinking about real life topics and how they apply to what I learn in the lab.
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