The Methanogens

Only the Methanosarcinales (e.g., Methanosarcina barkeri ) can reduce other substrates to CH4 according to the following equations:

Methanogenic organisms gain energy by using H2 to reduce CO2 to CH4. These organisms can also decarboxylate acetate to CO2 and CH4. Methane formation represents the terminal portion of a complex series of anaerobic reactions that occur in nature and involve a number of organisms that degrade biopolymers such as cellulose, starch, or proteins to acetate, H2, and CO2 . Conversion of complex organic material to these simple products requires the action of both primary and secondary fermenters from the clostridia and other anaerobic organisms. Primary fermenters can yield acetate, H2, and CO2. Other products require additional degradation by the secondary fermenters. Methanogenic microorganisms conduct the last portion of the conversion sequence to yield CH4 as a final product. Methanogens belong to the archaea (archaebacteria). Most methanogens can produce CH4 from H2 and CO2 as shown in the first equation below.

The major genera are listed in Table 12-1.

 Methanogens are obligate anaerobic Archaea that produce energy from the biosynthesis of methane. These lithotrophic microorganisms are widely distributed in oxygen-free environments and participate actively in the carbon cycle. Indeed, methanogenesis plays a major role in the last step of the anoxic degradation of organic substances, transforming acetate, CO(2), and H(2) to methane. The vast majority of the known methanogens are classified as hydrogenotrophic because they use principally H(2) as the electron donor to drive the reduction of CO(2). Unlike many other cultured Archaea, many methanogens thrive in neutral pH, low salinity, and temperate environments.

 Methanogens can be used to produce methane (aka natural gas, biogas) from biomass and degrade and detoxify agricultural, municipal, and industrial wastes. Methane is also a big contributor to global warming, which is why a better understanding of how methanogenesis works is needed so that we can use methane as a renewable source of energy and limit its greenhouse gas effect

 Although most methanogens have a pH optima near neutral, there are some methanogens that live in extreme pH environments. Methanogenesis has been shown to occur at low pH's (pH=3.0), but the optimal pH value is near 6.0.

Methanogens thrive in anaerobic environments rich in organic matter: the rumen and intestinal system of animals, freshwater and marine sediments, swamps and marshes, hot springs, anaerobic sludge digesters, and even within anaerobic protozoa. Methanogens often are of ecological significance. The rate of methane production can be so great that bubbles of methane will sometimes rise to the surface of a lake or pond. Rumen methanogens are so active that a cow can belch 200 to 400 liters of methane a day

 Methanogens are autotrophic archaebacteria that use anaerobic respiration for ATP synthesis. Methanogens use CO2 taken up from their growth environment as the carbon substrate for growth. They use some CO2 as the ultimate oxidizing agent of an electron transport chain which, by a chemiosmotic mechanism, maintains a transmembrane electrochemical ion gradient which powers ATP production.

 The reducing agent that drives the electron transport chain is hydrogen also taken up from the growth environment. This hydrogen is the waste end product of the metabolism of other, heterotrophic microorganisms. Methanogens use this hydrogen and this process maintains a lowered hydrogen partial pressure in the reticulo-rumen. Some of the hydrogen producing heterotrophic microorganisms show altered patterns of metabolism because of methanogen usage of the hydrogen they produce. This process lies at the heart of some of the best characterised syntrophic relations seen in the reticulo-rumen and and is referred to as "interspecies hydrogen transfer"

 This phenomenon is important because hydrogen utilization by the methanogens reduces the hydrogen partial pressure of the reticulorumen and this alters the pattern of metabolism of syntrophic hydrogen "donor" partner species. This is due to these organisms having hydrogen-sensitive hydrogenases. Raising or lowering the partial pressure of hydrogen in their growth environment determines their fermentation pathways and thus affects their ATP production which affects their growth. As we shall see further on in this section when studying the fermentation patterns of Ruminococcus flavifaciens a lowering of the partial pressure of hydrogen by interspecies hydrogen transfer allows enhanced growth of the syntrophic hydrogen "donor" species. The equation shows the reduction of CO2 by H2 to produce methane. This redox reaction sustains anaerobic respiration which allows the production of ATP.

The methane produced by reduction of the carbon dioxide is lost from the reticulorumen by eructation. It is a waste of feed carbon because the rumen does not have methanotrophic bacteria and the host ruminant can not utilize this gas. Methane lost in this way is one reason why methanogens contribute to lowered food conversion efficiency of the host ruminant.

Methanogenic archaea are potentially of great practical importance since methane is a clean-burning fuel and an excellent energy source.

 Methanogens are strict anaerobes that obtain energy by converting CO2, H2, formate, methanol, acetate, and other compounds to either methane or methane and CO2. They are autotrophic when growing on H2 and CO2.

This is the largest group of archaea.

There are five orders (Methanobacteriales, Methanococcales, Methanomicrobiales, Methanosarcinales, and Methanopyrales) and 26 genera, which differ greatly in overall shape, 16S rRNA sequence,cell wall chemistry and structure, membrane lipids, and other features.

Metabolism
Most methanogens can grow on CO2 and H2 as their sole energy source. coenzyme bound C1-intermediates Methanofuran (MFR), tetrahydromethanopterin (H4MPT), and coenzyme M (H-S-CoM). Other key coenzymes are F420 and N-7mercaptoheptanoyl-O-phospho-Lthreonine (H-S-HTP).

their metabolism is unusual. These procaryotes contain several unique cofactors:

tetrahydromethanopterin (H4MPT), coenzyme M methanofuran (MFR),

(2-mercaptoethanesulfonic acid), coenzyme F420, and coenzyme F430 The first three cofactors bear the C1 unit when CO2 is reduced to CH4. F420 carries electrons and hydrogens, and F430 is a nickel tetrapyrrole serving as a cofactor for the enzyme methyl-CoM methylreductase

These procaryotes contain several unique cofactors:

tetrahydromethanopterin (H4MPT), coenzyme M methanofuran (MFR),

(2-mercaptoethanesulfonic acid), coenzyme F420, and coenzyme F430 The first three cofactors bear the C1 unit when CO2 is reduced to CH4. F420 carries electrons and hydrogens, and F430 is a nickel tetrapyrrole serving as a cofactor for the enzyme methyl-CoM methylreductase

1. 4 CH3OH 3 CH4 + CO2 + 2 H20

2. CH3OH +H2 CH4 + H2O 3. 4 CH3NH2 + 2 H2O 3 CH4 + CO2 + 4 NH3 4. 2 (CH3)2NH + 2 H2O 3 CH4 + CO2 + 2 NH3 5. 4 (CH3)3N + 6 H2O 9 CH4 + 3 CO2 + 4 NH3 6. 2 (CH3)2S + 2 H2O 3 CH4 + CO2 + 2 H2S

G -106
-112.5 -76.7 -74.8 -75.8 -52.1

Energy-yielding reactions used by methylotrophic methanogens. One of the more common reactions, and the reaction showed above, is the 9th one down. While the last reaction involving methane is the most favorable, methane is not as readily available as carbon dioxide.

7. 4 (CH3)SH + 2 H20 3 CH4 + CO2 + 4 H2S 8.(CH3)SH + H2 CH4 +H2S

-51 8. -69.3

9. 4 H2 + CO2 CH4 + 2 H2O

-130.4

10. CH3COO- + H+ CH4 + CO2 -36 11. 4 CO + 2 H20 CH4 + 3 CO2 211

 Methanogenic archaea have an unusual type of metabolism because they use H2 + CO2, formate, methylated C1 compounds, or acetate as energy and carbon sources for growth. The methanogens produce methane as the major end product of their metabolism in a unique energy-generating process. The organisms received much attention because they catalyze the terminal step in the anaerobic breakdown of organic matter under sulfate-limiting conditions and are essential for both the recycling of carbon compounds and the maintenance of the global carbon flux on Earth. Furthermore, methane is an important greenhouse gas that directly contributes to climate changes and global warming. Hence, the understanding of the biochemical processes leading to methane formation are of major interest. This review focuses on the metabolic pathways of methanogenesis that are rather unique and involve a number of unusual enzymes and coenzymes. It will be shown how the previously mentioned substrates are converted to CH4 via the CO2-reducing, methylotrophic, or aceticlastic pathway.

 All catabolic processes finally lead to the formation of a mixed disulfide from coenzyme M and coenzyme B that functions as an electron acceptor of certain anaerobic respiratory chains. Molecular hydrogen, reduced coenzyme F420, or reduced ferredoxin are used as electron donors. The redox reactions as catalyzed by the membrane-bound electron transport chains are coupled to proton translocation across the cytoplasmic membrane. The resulting electrochemical proton gradient is the driving force for ATP synthesis as catalyzed by an A1A0-type ATP synthase.

 Other energy-transducing enzymes involved in methanogenesis are the membrane-integral methyltransferase and the formylmethanofuran dehydrogenase complex. The former enzyme is a unique, reversible sodium ion pump that couples methylgroup transfer with the transport of Na+ across the membrane. The formylmethanofuran dehydrogenase is a reversible ion pump that catalyzes formylation and deformylation of methanofuran. Furthermore, the review addresses questions related to the biochemical and genetic characteristics of the energy-transducing enzymes and to the mechanisms of ion translocation.

 Some examples of rod shaped cells include Methanobacterium spp. and Methanopyrus kandleri. Examples of the coccoid methanogens include species from Methanococcus and Methanosphaera to name a few. Methanoculleus and Methanogenium are coccoid as well but are irregularly shaped, possibly due to S-layers not being so strongly bonded like other wall structures. Methanogens are not just limited to these shapes, but include a plate shaped genus Methanoplanus, Methanospirillum that are long thin spirals, and Methanosarcina that are cluster of round cells.

Ref Prescott pg 612