In the absence of oxygen, bacteria use fermentation (substrate-level phosphorylation with organic electron acceptors) or anaerobic respiration (electron transport with inorganic acceptors like nitrate). These pathways regenerate NAD+ and generate ATP, enabling growth in anoxic environments such as gut, sediment, and aquatic systems.
Culture bacteria anaerobically and measure lactate, ethanol, or other fermentation products. Compare growth rates under aerobic vs. anaerobic conditions.
Anaerobic respiration is not the same as fermentation—anaerobic respiration still uses an electron transport chain. Not all bacteria can ferment; many strictly require oxygen or an alternate electron acceptor.
From your study of microbial fermentation, you know the basic problem: when cells oxidize glucose through glycolysis, they reduce NAD+ to NADH, and they need a way to regenerate NAD+ to keep glycolysis running. Aerobic organisms solve this by passing electrons from NADH through an electron transport chain to oxygen, the ultimate electron acceptor. But many environments — deep sediments, waterlogged soils, the interior of the mammalian gut — contain little or no oxygen. Bacteria thriving in these habitats have evolved two fundamentally different strategies for coping, and the distinction between them is one of the most important concepts in microbial metabolism.
Fermentation is the simpler strategy. Instead of using an electron transport chain at all, fermentative bacteria transfer electrons from NADH directly to an organic molecule — typically pyruvate or a derivative of it. Lactic acid bacteria reduce pyruvate to lactate; *Saccharomyces* (yeast, though not a bacterium) converts it to ethanol and CO₂; other organisms produce butyrate, propionate, or mixed acids. The sole purpose of these reactions is to regenerate NAD+ so that glycolysis can continue generating ATP through substrate-level phosphorylation. The organic end products still contain substantial chemical energy, which is why fermentation yields far less ATP per glucose molecule (typically just 2 ATP) compared to aerobic respiration (up to 38). The diverse fermentation products are not waste in an ecological sense — they feed other organisms in the community and form the basis of food webs in anaerobic environments.
Anaerobic respiration is a more sophisticated strategy that retains the electron transport chain but substitutes a different terminal electron acceptor in place of oxygen. Denitrifying bacteria use nitrate (NO₃⁻), reducing it stepwise to nitrite, nitric oxide, nitrous oxide, and finally N₂ gas — a process critical to the global nitrogen cycle. Sulfate-reducing bacteria use sulfate (SO₄²⁻), producing hydrogen sulfide (H₂S), the compound responsible for the rotten-egg smell of anoxic mud. Others use iron(III), manganese(IV), or even carbon dioxide as electron acceptors. Because anaerobic respiration uses a proton motive force and an electron transport chain, it generates significantly more ATP than fermentation — though still less than aerobic respiration, because these alternative acceptors have lower reduction potentials than oxygen.
The ecological significance of these pathways is enormous. Fermentation and anaerobic respiration drive biogeochemical cycling of nitrogen, sulfur, and carbon in oxygen-depleted habitats that cover vast areas of the planet. In the human gut, anaerobic bacteria outnumber aerobic ones by orders of magnitude, and their fermentation products — particularly short-chain fatty acids like butyrate — serve as major energy sources for intestinal epithelial cells and play roles in immune regulation. Understanding whether an organism ferments or respires anaerobically also has direct clinical relevance: it determines which metabolic products accumulate in an infection, how the organism will behave in culture, and which antibiotics (like aminoglycosides, which require aerobic uptake) will be ineffective against it.