Fermentation, mitochondria and regulation

Learning objectives

  1. Identify what cellular metabolic pathways can operate in the absence of respiration
  2. Predict how cellular pathways respond to the absence of terminal electron acceptors
  3. Compare and contrast how NAD+ is regenerated in respiration and fermentation
  4. Compare and contrast eukaryotic and prokaryotic metabolic pathways
  5. Cite evidence to support the endosymbiotic origin of mitochondria


Some cells make ATP solely via substrate-level phosphorylation, either because they lack the electron transport chain, or because suitable terminal electrons acceptors are unavailable. They use glycolysis to make 2 ATP and 2 pyruvates from a molecule of glucose, plus 2 NADH. However, such cells cannot continue running glycolysis indefinitely because they quickly run out of NAD+, when all available NAD+ has been reduced to NADH. In respiring cells, NADH dumps electrons to the electron transport chain and regenerates NAD+. In the absence of respiration, this method of regenerating NAD+ is not available.

Fermentation reactions reduce pyruvate with electrons from NADH to regenerate NAD+ (opposite of pyruvate oxidation). These reactions produce ethanol in yeast, and lactic acid in mammalian cells (muscle cells under oxygen deficit and most tumor cells – see Warburg effect below).

Ethanol fermentation regenerates NAD+ from NADH by releasing CO2 from pyruvate and reducing the two-carbon acetaldehyde molecule to ethanol. Image by Davidcarmack, retrieved from Wikimedia commons.

Mammalian cells regenerate NAD+ by reducing pyruvate to lactic acid. Image by sjantoni, retrieved from Wikimedia commons

Fermentation reactions occur in the cytoplasm of both prokaryotic and eukaryotic cells. In the absence of oxygen, pyruvate does not enter the mitochondria in eukaryotic cells.

Cancer and lactic acid fermentation – the Warburg effect

The Nobel Prize winner Otto Warburg observed that many, and perhaps most, cancer cells derive most of their energy from glycolysis and lactic acid fermentation, even when oxygen is plentiful (see review by Liberti and Locasale, 2016). Several explanations have been proposed. One is that cancer cells can promote biosynthesis and cell growth by NOT respiring organic carbon to CO2, and using the organic carbon to build cellular biomolecules, instead. Another hypothesis is that ramping up glycolysis allows tumor cells to out-compete normal cells or immune system cells for glucose. A third hypothesis is that lactic acid secretion causes changes in the tumor cells’ environment that favors tumor cell growth and spread.

Energy metabolism in eukaryotes vs prokaryotes

In prokaryotic cells, all the metabolic pathways occur in the cytoplasm, except for chemiosmosis and oxidative phosphorylation, which occur on the plasma membrane. Prokaryotic cells are capable of anaerobic respiration using alternative electron acceptors such as nitrate and sulfate, although they prefer oxygen as the terminal electron acceptor to drive chemiosmotic ATP synthesis. In the absence of any suitable electron acceptor, they use fermentation pathways.

In eukaryotic cells, glycolysis and fermentation reactions occur in the cytoplasm. The remaining pathways, starting with pyruvate oxidation, occur in the mitochondria. Most eukaryotic mitochondria can use only oxygen as the terminal electron acceptor for respiration. In the presence of oxygen, pyruvate enters the mitochondrial matrix and is oxidized to acetyl-CoA, and then to CO2 via the citric acid cycle. The electron transport chain and ATP synthase are located on the mitochondrial inner membrane.

Energy metabolism in eukaryotic cells, from Wikipedia. This is a highly condensed illustration. Students should identify the various compounds and talk through (explain) this diagram.

In the absence of oxygen, pyruvate does not enter mitochondria, but instead undergoes fermentation to either lactic acid or ethanol.

The video below reviews what happens when cells cannot respire to create ATP:

Endosymbiont theory for origin of mitochondria

The location of the electron transport chain on the inner mitochondrial membrane, and the pyruvate oxidation and citric acid cycle in the mitochondrial matrix, makes sense in light of the endosymbiont theory for the origin of mitochondria. These locations correspond to the plasma membrane and cytoplasm of the aerobic bacterial endosymbiont, most likely an alpha-proteobacterium, that was the ancestor of mitochondria. The outer mitochondrial membrane derived from the endosomal membrane that originally engulfed the endosymbiont.

Further evidence to support the endosymbiont theory is that mitochondria have their own DNA, in the form of a circular chromosome that is topologically like bacterial chromosomes. The sequence of the mitochondrial DNA most closely resembles the sequences of genes in alpha-proteobacteria. Mitochondrial ribosomes are structurally more similar to bacterial ribosomes than to eukaryotic ribosomes. Mitochondria reproduce in eukaryotic cells by fission, again resembling bacterial cell division.


Liberti MV, Locasale JW (2016) The Warburg Effect: How Does It Benefit Cancer Cells? Trends Biochem Sci 41:211 – 218 DOI: http://dx.doi.org/10.1016/j.tibs.2015.12.001

A couple of music videos for your study breaks:

Industry, Innovation, and Infrastructure

UN Sustainable Development Goal (SDG) 9: Industry, Innovation, and Infrastructure – Fermentation allows for ATP generation when oxygen is not available. This is important in anaerobic environments, such as the bottom of lakes and ponds, where organisms such as bacteria and yeast survive by carrying out fermentation. Studying such organisms has lead to insights on the factors that promote fermentation as a metabolic process, as well as the factors that preserve ecosystem health through the utilization of different metabolic pathways under specific conditions. Providing organisms with the conditions that favor fermentation, or selectively remove the ability to ferment, also has industrial applications in the production of bread, beer, and wine. Here, toggling the ability of organisms to ferment creates characteristic molecules (like flavors and alcohols) and preserves food safety by limiting the growth of disease-causing microorganisms.

4 Responses to Fermentation, mitochondria and regulation

  1. Jung Choi says:

    A good, brief summary of respiration in eukaryotes in this Biology Crash Course video on ATP and respiration by Hank Green: http://www.youtube.com/watch?v=00jbG_cfGuQ

  2. Abhinav says:

    I have a question on the formation of lactate. When I compare the two lactate molecules with the 2 pyruvates, there are 4 additional H atoms. So, does each NADH lose 2 Hydrogen atoms, or is there another source of H atoms?

    • Jung Choi says:

      The additional H atoms are protons picked up from dissociation of water. Water spontaneously dissociates into H+ and OH- ions; at neutral pH (pH 7), the concentration of H+ is 10exp-7 Molar. The complete equation for reduction of pyruvate is: pyruvate + NADH + H+ –> lactate + NAD+

  3. Jung Choi says:

    CNN has a story about a couple who learned that their child has a mitochondrial disease: http://www.cnn.com/2012/09/21/health/mitochondrial-disease-martin-family/index.html

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