The contraction of skeletal muscles is an energy-requiring process. In order to perform the mechanical work of contraction, actin and myosin utilize the chemical energy of the molecule adenosine triphosphate (ATP). ATP is synthesized in muscle cells from the storage polysaccharide glycogen, a complex carbohydrate composed of hundreds of covalently linked molecules of glucose (a monosaccharide or simple carbohydrate). In a working muscle, glucose is released from the glycogen reserves and enters a metabolic pathway called glycolysis, a process in which glucose is broken down and the energy contained in its chemical bonds is harnessed for the synthesis of ATP. The net production of ATP depends on the level of oxygen reaching the muscle. In the absence of oxygen (anaerobic conditions), the products of glycolysis are converted to lactic acid, and relatively little ATP is produced. In the presence of oxygen (aerobic conditions), the products of glycolysis enter a second pathway, the citric acid cycle, and a large amount of ATP is synthesized by a process called oxidative phosphorylation.

In addition to carbohydrates, fats supply a significant amount of energy for working muscles. Fats are stored in the body as triglycerides (also called triacylglycerols). A triglyceride is composed of three fatty acid molecules (nonpolar hydrocarbon chains with a polar carboxyl group at one end) bound to a single glycerol molecule. If the fat deposits are required for energy production, fatty acids are released from the triglyceride molecules in a process called fatty acid mobilization. The fatty acids are broken down into smaller molecules that can enter the citric acid cycle for the synthesis of ATP by oxidative phosphorylation. Therefore, the utilization of fats for energy requires the presence of oxygen.

An important protein of muscle cells is the oxygen-binding protein myoglobin. Myoglobin takes up oxygen from the blood (transported by the related oxygen-binding protein hemoglobin) and stores it in the muscle cells for oxidative metabolism. The structure of myoglobin includes a nonprotein group called the heme ring. The heme ring consists of a porphyrin molecule bound to an iron (Fe) atom. The iron atom is responsible for the binding of oxygen to myoglobin and has two possible oxidation states: the reduced, ferrous form (Fe²⁺) and the oxidized, ferric form (Fe³⁺). In the Fe²⁺ state iron is able to bind oxygen (and other molecules). However, oxidation of the iron atom to the Fe³⁺ state prevents oxygen binding.

pH changes

Normally, after death, muscle becomes more acidic (pH decreases). When an animal is bled after slaughter (a process known as exsanguination), oxygen is no longer available to the muscle cells, and anaerobic glycolysis becomes the only means of energy production available. As a result, glycogen stores are completely converted to lactic acid, which then begins to build up, causing the pH to drop. Typically, the pH declines from a physiological pH of approximately 7.2 in living muscle to a postmortem pH of approximately 5.5 in meat (called the ultimate pH).

Protein changes

When the energy reserves are depleted, the myofibrillar proteins, actin and myosin, lose their extendability, and the muscles become stiff. This condition is commonly referred to as rigor mortis. The time an animal requires to enter rigor mortis is highly dependent on the species (for instance, cattle and sheep take longer than hogs), the chilling rate of the carcass from normal body temperature (the process is slower at lower temperatures), and the amount of stress the animal experiences before slaughter.

Eventually the stiffness in the muscle tissues begins to decrease owing to the enzymatic breakdown of structural proteins (i.e., collagen) that hold muscle fibres together. This phenomenon is known as resolution of rigor and can continue for weeks after slaughter in a process referred to as aging of meat. This aging effect produces meats that are more tender and palatable.