The actual production of ATP in cellular respiration takes place through the process of chemiosmosis. Chemiosmosis involves the pumping of protons through special channels in the membranes of mitochondria from the inner to the outer compartment. The pumping establishes a proton gradient. After the gradient is established, protons pass down the gradient through particles designated F1. In these particles, the energy of the protons generates ATP, using ADP and phosphate ions as the starting points.

The energy production of cellular respiration is substantial. Most biochemists agree that 36 molecules of ATP can be produced for each glucose molecule during cellular respiration as a result of the Krebs cycle reactions, the electron transport system, and chemiosmosis. Also, two ATP molecules are produced through glycolysis, so the grand total is 38 molecules of ATP. These ATP molecules may then be used in the cell for its needs. However, the ATP molecules cannot be stored for long periods of time, so cellular respiration must constantly continue in order to regenerate the ATP molecules as they are used. Each ATP molecule is capable of releasing 7.3 kilocalories of energy per mole.