Blog Archive

Thursday, September 4, 2025

Methanogenesis: Part 3: Fingerprinting Methane by Isotopes Reveals Sources and CRISPR Gives Insights into Enzyme Mechanisms for Methanogenesis

     I am certainly no expert, but I think I should better explain methanogenesis for this multiple-part post. Methanogenesis refers to biologically produced methane and is also referred to as bio-methanization. In my oil & gas studies and work, I came to understand the two main sources of subsurface methane generation: biogenic and thermogenic. Methane in drilled natural gas fields may be of either type or a combination, but the bulk of it is thermogenic, produced by the heat and pressure in deeply buried rock that is rich in organic matter, typically decomposed algal matter. The PT conditions influence the chemical “cracking” of heavier hydrocarbons into methane. Dry natural gas occurs when the “wetter” liquids that make up crude oil are used up in the process. Dry natural gas occurs where hydrocarbons are said to be thermally mature. There is also abiotic methane produced by the interaction of seawater and magmatic olivine in the process known as serpentinization. This has recently been shown to be more prevalent at deep-sea trenches than thought. According to a 2019 release from Woods Hole Oceanographic Institution:

“…seawater, moving through the deep oceanic crust, is trapped in magma-hot olivine.  As the mineral cools, the water trapped inside undergoes a chemical reaction, a process called serpentinization that forms hydrogen and methane. The authors demonstrate that in otherwise inhospitable environments, just two ingredients—water and olivine—can form methane.”   

     According to Wikipedia:

Methanogenesis or biomethanation is the formation of methane coupled to energy conservation by microbes known as methanogens. It is the fourth and final stage of anaerobic digestion. Organisms capable of producing methane for energy conservation have been identified only from the domain Archaea, a group phylogenetically distinct from both eukaryotes and bacteria, although many live in close association with anaerobic bacteria. The production of methane is an important and widespread form of microbial metabolism. In anoxic environments, it is the final step in the decomposition of biomass. Methanogenesis is responsible for significant amounts of natural gas accumulations, the remainder being thermogenic.”




     Marine sediments are an important habitat for methane-generating microbial communities. In the most common pathway, the microbes consume acetate, which comprises two-thirds of global methane production. The abbreviated reactions are CO2 + 4 H2 → CH4 + 2 H2O and CH3COOH → CH4 + CO2. The pathways are shown in more detail below.




     Both carbon and hydrogen isotopes can be used to fingerprint methane. An August 2025 paper in Science explores the main enzyme in methanogenesis, methyl–coenzyme M reductase (MCR), and how modulating it with CRISPR alters the isotopic composition of microbial methane, showing that a similar modulation pathway exists in nature. Jonathan Gropp, the paper’s lead author, notes:

“…for methane, large uncertainties in fluxes exist -- within tens of percents for some of the fluxes -- that challenge our ability to precisely quantify the relative importance and changes in time of the sources. To quantify the actual sources of methane, you need to really understand the isotopic processes that are used to constrain these fluxesMicrobes respond to the environment by manipulating their gene expression, and then the isotopic compositions change as well. This should cause us to think more carefully when we analyze data from the environment."

     Co-author Dipti Nayak, UC Berkeley assistant professor of molecular and cell biology, added:

"It is well understood that methane levels are rising, but there is a lot of disagreement on the underlying cause. This study is the first time the disciplines of molecular biology and isotope biogeochemistry have been fused to provide better constraints on how the biology of methanogens controls the isotopic composition of methaneI think what's unique about the paper is, we learned that the isotopic composition of microbial methane isn't just based on what methanogens eat. What you 'eat' matters, of course, but the amount of these substrates and the environmental conditions matter too, and perhaps more importantly, how microbes react to those changes.”

     Geochemist and co-author Daniel Stolper, UC Berkeley associate professor of earth and planetary science, explained isotopic signatures and variation in methanogenesis pathways:

"Over the last 70 years, people have shown that methane produced by different organisms and other processes can have distinctive isotopic fingerprints. Natural gas from oil deposits often looks one way. Methane made by the methanogens within cow guts looks another way. Methane made in deep sea sediments by microorganisms has a different fingerprint. Methanogens can consume or 'eat', if you will, a variety of compounds including methanol, acetate or hydrogen; make methane; and generate energy from the process. Scientists have commonly assumed that the isotopic fingerprint depends on what the organisms are eating, which often varies from environment to environment, creating our ability to link isotopes to methane origins."

     The researchers used CRISPR to reduce the activity of the enzyme and found that when they did, the isotopic composition of the methane changed. They also discovered that changes in the availability of food sources for the microbes result in changes in gene expression that lead to changes in isotopic signatures. These archaean microbes consume acetate (essentially vinegar), methanol (the simplest alcohol), or molecular hydrogen (H2)  and produce methane, CH4, with a ratio of hydrogen and carbon isotopes different from the ratios observed in the environment. They found that the microbes, which normally get hydrogen from what they consume, can also get it from the water in the environment if their food source becomes scarce.




     According to Science Daily and the authors, using CRISPR in similar isotope/enzyme studies can help increase understanding:

Beyond this study, the CRISPR technique for tuning production of enzymes in methanogens could be used to manipulate and study isotope effects in other enzyme networks broadly, which could help researchers answer questions about geobiology and the Earth's environment today and in the past.”

"This opens up a pathway where modern molecular biology is married with isotope-geochemistry to answer environmental problems," Stolper said. "There are an enormous number of isotopic systems associated with biology and biochemistry that are studied in the environment; I hope we can start looking at them in the way molecular biologists now are looking at these problems in people and other organisms -- by controlling gene expression and looking at how the stable isotopes respond."

     Another potential benefit could be using the knowledge gained to one day use it to reduce the methane output to the atmosphere, but this is likely far off.

 

 

    

References:

 

Methanogenesis. Wikipedia. Methanogenesis - Wikipedia

Scientists just found a hidden factor behind Earth’s methane surge: Using CRISPR to dial down enzyme helps to understand the isotope signatures of methane from different environments. Sceince Daily. University of California at Berkeley. August 17, 2025. Scientists just found a hidden factor behind Earth’s methane surge | ScienceDaily

Modulation of methyl–coenzyme M reductase expression alters the isotopic composition of microbial methane. Jonathan Gropp, Markus Bill, Max K. Lloyd, Rebekah A. Stein, Dipti D. Nayak, and Daniel A. Stolper. Science. 14 Aug 2025. Vol 389, Issue 6761. pp. 711-715. Modulation of methyl–coenzyme M reductase expression alters the isotopic composition of microbial methane | Science

Origin of Massive Methane Reservoir Identified. Woods Hole Oceanographic Institution. August 20, 2019. Origin of Massive Methane Reservoir Identified – Woods Hole Oceanographic Institution

No comments:

Post a Comment

       This is an interesting blog by a senior geologist specializing in CCS and decarbonization. I have attended one of Jason’s excellent ...