Lesson Content

Inquire: Glycolysis


Glycolysis is the first pathway used in the breakdown of glucose to extract energy. Because it is used by nearly all organisms on earth, it must have evolved early in the history of life. Glycolysis consists of two parts. The first part prepares the six-carbon ring of glucose for separation into two three-carbon sugars. Energy from ATP is invested into the molecule during this step, in order to energize the separation. The second half of glycolysis extracts ATP and high-energy electrons from hydrogen atoms and attaches them to NAD+. Two ATP molecules are invested in the first half and four ATP molecules are formed during the second half. This produces a net gain of two ATP molecules per glucose molecule for the cell.


Big Question

What is the purpose of glycolysis?

Watch: The Breakdown of Glucose

Read: Glycolysis


Even energy-releasing exergonic reactions require a small amount of activation energy to proceed. However, consider endergonic reactions, which require much more energy input because their products have more free energy than their reactants. Within the cell, where does energy to power such reactions come from? The answer lies with an energy-supplying molecule called adenosine triphosphate, or ATP. ATP is a small, relatively simple molecule, but its bonds contain the potential for a quick burst of energy that can be harnessed to perform cellular work. This molecule can be thought of as the primary energy currency of cells in the same way that money is the currency that people exchange for things they need. ATP is used to power the majority of energy-requiring cellular reactions.

ATP in Living Systems

A living cell cannot store significant amounts of free energy. Excess free energy would result in an increase of heat in the cell, which would denature enzymes and other proteins, and thus destroy the cell. Rather, a cell must be able to store energy safely and release it for use only as needed. Living cells accomplish this using ATP, which can be used to fill any energy need of the cell. How? It functions as a rechargeable battery.

When ATP is broken down, usually by the removal of its terminal phosphate group, energy is released. This energy is used to do work by the cell, usually by the binding of the released phosphate to another molecule, thus activating it. For example, in the mechanical work of muscle contraction, ATP supplies energy to move the contractile muscle proteins.

ATP Structure and Function

At the heart of ATP is a molecule of adenosine monophosphate (AMP), which is composed of an adenine molecule bonded to both a ribose molecule and a single phosphate group.

DecorativeRibose is a five-carbon sugar found in RNA, and AMP is one of the nucleotides in RNA. The addition of a second phosphate group to this core molecule results in adenosine diphosphate (ADP); the addition of a third phosphate group forms adenosine triphosphate (ATP).

The addition of a phosphate group to a molecule requires a high amount of energy and results in a high-energy bond. Phosphate groups are negatively charged and thus repel one another when they are arranged in series, as they are in ADP and ATP. This repulsion makes the ADP and ATP molecules inherently unstable. The release of one or two phosphate groups from ATP, a process called hydrolysis, releases energy.


Nearly all energy used by living things comes to them in the bonds of the sugar, or glucose. Glycolysis is the first step in the breakdown of glucose to extract energy for cell metabolism. Many living organisms carry out glycolysis as part of their metabolism. Glycolysis is an anaerobic process (does not use oxygen) which takes place in the cytoplasm of most prokaryotic, and all eukaryotic, cells.

Glycolysis begins with the six-carbon, ring-shaped structure of a single glucose molecule and ends with two molecules of a three-carbon sugar called pyruvate. Glycolysis consists of two distinct phases. In the first part of the glycolysis pathway, energy is used to make adjustments so that the six-carbon sugar molecule can be split evenly into two three-carbon pyruvate molecules. In the second part of glycolysis, ATP and nicotinamide-adenine dinucleotide (NADH) are produced.

DecorativeIf the cell cannot catabolize the pyruvate molecules further, it will harvest only two ATP molecules from one molecule of glucose. For example, mature mammalian red blood cells are only capable of glycolysis, which is their sole source of ATP. If glycolysis is interrupted, these cells would eventually die.

Outcomes of Glycolysis

Glycolysis begins with glucose and produces two pyruvate molecules, four new ATP molecules, and two molecules of NADH. If the cell cannot catabolize the pyruvate molecules further, it will harvest only two ATP molecules from one molecule of glucose. Mature mammalian red blood cells do not have mitochondria and thus are not capable of aerobic respiration — the process in which organisms convert energy in the presence of oxygen — and glycolysis is their sole source of ATP. If glycolysis is interrupted, these cells lose their ability to maintain their sodium-potassium pumps, and eventually, they die.

The last step in glycolysis will not occur if pyruvate kinase, the enzyme that catalyzes the formation of pyruvate, is not available in sufficient quantities. In this situation, the entire glycolysis pathway will proceed, but only two ATP molecules will be made in the second half. Thus, pyruvate kinase is a rate-limiting enzyme for glycolysis.

Reflect: What to Eat?


Since energy comes from the breakdown of glucose, which food group do you think would provide you with the most energy?

Expand: Adenosine Triphosphate


As its name suggests, adenosine triphosphate (ATP) is made up of adenosine bound to three phosphate groups. Adenosine is a nucleoside consisting of the nitrogenous base adenine and a five-carbon sugar, ribose. The three phosphate groups, in order of closest to furthest from the ribose sugar, are alpha, beta, and gamma. Together these chemical groups constitute an energy powerhouse. However, not all bonds within this molecule exist in a particularly high-energy state.

Both bonds that link the phosphates are equally high-energy bonds that, when broken, release sufficient energy to power a variety of cellular reactions and processes. These high-energy bonds are the bonds between the second and third phosphate groups and between the first and second phosphate groups. These bonds are “high-energy” because the products of such bond breaking — adenosine diphosphate (ADP) and one inorganic phosphate group — have considerably lower free energy than the reactants, ATP and a water molecule. Because this reaction takes place using a water molecule, it is a hydrolysis reaction. In other words, ATP hydrolyzes into ADP in the following reaction:

ATP+H2O→ADP+Pi+free energy

Like most chemical reactions, ATP to ADP hydrolysis is reversible. The reverse reaction regenerates ATP from ADP + Pi. Cells rely on ATP regeneration just as people rely on regenerating their spent money through some sort of income. Since ATP hydrolysis releases energy, ATP regeneration must require an input of free energy. This equation expresses ATP formation:

ADP+Pi+free energy→ATP+H2O

ATP is a highly unstable molecule. Unless quickly used to perform work, ATP spontaneously dissociates into ADP + Pi, and the free energy released during this process is lost as heat. Cells couple the ATP hydrolysis’ exergonic reaction allowing them to proceed. One example of energy coupling using ATP involves a transmembrane ion pump that is extremely important for cellular function. This sodium-potassium pump drives sodium out of the cell and potassium into the cell.

A large percentage of a cell’s ATP powers this pump, because cellular processes bring considerable sodium into the cell and push potassium out of it. The pump works constantly to stabilize cellular concentrations of sodium and potassium. In order for the pump to turn one cycle, one ATP molecule must hydrolyze. When ATP hydrolyzes, its gamma phosphate does not simply float away but actually transfers onto the pump protein. Scientists call this process in which a phosphate group binds to a molecule phosphorylation.

DecorativeAs with most ATP hydrolysis cases, a phosphate from ATP transfers onto another molecule. In a phosphorylated state, the Na+/K+ pump has more free energy and is triggered to undergo a conformational change. This change allows it to release Na+ to the cell’s outside. It then binds extracellular K+, which, through another conformational change, causes the phosphate to detach from the pump. This phosphate release triggers the K+ to release to the cell’s inside. Essentially, the energy released from the ATP hydrolysis couples with the energy required to power the pump and transport Na+ and K+ ions. ATP performs cellular work using this basic form of energy coupling through phosphorylation.

Often during cellular metabolic reactions, such as nutrient synthesis and breakdown, certain molecules must alter slightly in their conformation to become substrates for the next step in the reaction series. An example of this is during the very first steps of cellular respiration, when a sugar glucose molecule breaks down in the process of glycolysis. In the first step, ATP is required to phosphorylze glucose, creating a high-energy but unstable intermediate. This phosphorylation reaction powers a conformational change that allows the phosphorylated glucose molecule to convert to the phosphorylated sugar fructose. Fructose is a necessary intermediate for glycolysis to move forward. Here, ATP hydrolysis’ exergonic reaction couples with the endergonic reaction of converting glucose into a phosphorylated intermediate in the pathway. Once again, the energy released by breaking a phosphate bond within ATP was used for phosphorylyzing another molecule, creating an unstable intermediate and powering an important conformational change.

Check Your Knowledge

Use the quiz below to check your understanding of this lesson’s content. You can take this quiz as many times as you like.

Lesson Resources

Lesson Toolbox

Additional Resources and Readings

Glycolysis: An Overview

A video giving a better understanding of glucose breakdown by glycolysis

Glycolysis! (Mr. W’s Music Video)

A music video to help you understand and remember glycolysis

ATP Adenosine Triphosphate

A short clip explaining ATP

Lesson Glossary


AJAX progress indicator
  • adenosine triphosphate (ATP)
    a very high-energy molecule that is the primary energy currency of all cells
  • aerobic respiration
    process in which organisms convert energy in the presence of oxygen
  • anaerobic
    process that does not use oxygen
  • endergonic reactions
    a reaction that requires energy to be driven
  • exergonic reactions
    a reaction where energy is released
  • glycolysis
    process of breaking glucose into two three-carbon molecules with the production of ATP and NADH
  • hydrolysis
    the release of one or two phosphate groups from ATP, which releases energy
  • phosphorylation
    a biochemical process that involves the addition of phosphate to an organic compound
  • pyruvate
    three-carbon sugar that can be decarboxylated and oxidized to make acetyl CoA, which enters the citric acid cycle under aerobic conditions; the end product of glycolysis
  • pyruvate kinase
    the enzyme that catalyzes the formation of pyruvate

License and Citations

Content License

Lesson Content:

Authored and curated by Jill Carson for The TEL Library. CC BY NC SA 4.0

Adapted Content:

Title: Biology – 7.2 Glycolysis – First Half of Glycolysis (Energy-Requiring Steps); OpenStax CNX. License: CC BY 4.0

Title: Biology – 6.4 Adenosime Triphosphate, OpenStax CNX. License: CC BY 4.0

Title: Biology – 4.2 Glycolysis – ATP in Living Systems; Glycolysis; OpenStax CNX. License: CC BY 4.0

Media Sources

Decorative1,3-BPG-3-PGakane700Wikimedia CommonsPublic Domain
DecorativeGSK3 phosphorylationHyouWikimedia CommonsPublic Domain
Decorativeglucose moleculeOpenStaxOpenStaxCC BY 4.0
DecorativeATPOpenStaxOpenStaxCC BY 4.0