# Glycolysis and Cellular Respiration

### Introduction

#### What is Cellular Respiration?

Cellular respiration allows organisms to use (release) energy stored in the chemical bonds of glucose (C6H12O6). The energy in glucose is used to produce ATP. Cells use ATP to supply their energy needs. Cellular respiration is therefore a process in which the energy in glucose is transferred to ATP.

In respiration, glucose is oxidized and thus releases energy. Oxygen is reduced to form water.

The carbon atoms of the sugar molecule are released as carbon dioxide (CO2).

The complete breakdown of glucose to carbon dioxide and water requires two major steps: 1) glycolysis and 2) aerobic respiration.  Glycolysis produces two ATP. Thirty-four more ATP are produced by aerobic pathways if oxygen is present.

In the absence of oxygen, fermentation reactions produce alcohol or lactic acid but no additional ATP.

### Review of Electron Carriers

$\mathrm{NAD^+ + 2H \rightarrow NADH + H^+}$

$\mathrm{FAD + 2H \rightarrow FADH_2}$

### Glycolysis

During glycolysis, glucose (C6) is broken down to two molecules of pyruvate (C3).  (Note that compounds that end in "___ate" can be called "___ic acid". For example, lactate is lactic acid and malate is malic acid.)

Glycolysis occurs in the cytoplasm (cytosol) and does not require oxygen.

There are ten steps in glycolysis and each one is catalyzed by a specific enzyme. A brief summary of these reactions is presented here.

2 ATP molecules are used to phosphorylate and activate compounds that will eventually become converted to pyruvate (or pyruvic acid) (see diagram below).

Two hydrogen atoms are removed by NAD+ forming 2 NADH (see diagram).

Additional phosphorylation results in intermediate 3-carbon molecules with 2 phosphate groups.

Four ATP are produced by substrate-level phosphorylation. Recall that substrate-level phosphorylation is the production of ATP using energy from other high-energy compounds but without the use of the electron transport system in the mitochondria.

The net yield of ATP in glycolysis is 2 for each glucose molecule (2 are used but 4 are produced).

Some bacteria have alternative energy-producing reactions. Two of these are the pentose phosphate pathway and the Entner-Doudoroff pathway.

### Formation of Acetyl CoA

Pyruvate produced by glycolysis (see above) enters the mitochondrion by active transport and is converted to acetyl CoA as shown below. The remainder of the reactions of cellular respiration occur in the mitochondrion.

$\mathrm{pyruvate\: (C_3) \rightarrow acetyl\: CoA\: (C_2) + CO_2}$

A carbon atom is removed from each of the pyruvate molecules forming a two-carbon compound and CO2.

Coenzyme A is attached to each of the two-carbon compounds producing two acetyl CoA molecules.

### Citric Acid Cycle

The citric acid cycle can be summarized by either of the diagrams below. The diagram below occurs twice, once for each acetyl CoA.

Coenzyme A is removed when the two-carbon compound is attached to a four-carbon compound producing a six-carbon compound (citrate).

Each citrate molecule undergoes a series of reactions that removes 2 carbon atoms which are released as CO2. In addition, 3 NADH, 1 ATP, and 1 FADH2 are produced. In addition, the four-carbon compound that began the cycle is regenerated.

The diagram below also summarizes the citric acid cycle.

#### Mitochondrion structure

The inner membrane forms folds called cristae. These folds contain the carriers of the electron transport system.

Acetyl CoA formation and the citric acid cycle occur in the inner space called the matrix.

The space outside the inner membrane is the intermembrane space. The electron transport system pumps hydrogen ions (H+) into this space for oxidative phosphorylation.

#### Oxidative Phosphorylation

The electron transport system is found in the mitochondrion and chloroplast of eucaryotes and in the plasma membrane of procaryotes. It consists of a series of carrier molecules which pass electrons from a high-energy compound to a final low-energy electron acceptor. Energy is released during these oxidation-reduction reactions to produce ATP.

The discussion below applies to the mitochondria of eucaryotes.

NADH or FADH2 bring electrons to the electron transport system in the mitochondria.

The system contains membrane-bound electron carriers that pass electrons from one to another. When a carrier reduces another, some of the energy that is released as a result of that reduction is used to pump hydrogen ions across the membrane into the intermembrane space. The remaining energy is used to reduce the next carrier.

As a result of the electron transport system, hydrogen ions become concentrated in the intermembrane space. These concentrated ions contain energy much like a dam. The enzyme ATP synthase is able to use the energy of this osmotic gradient to produce ATP as the hydrogen ions move under osmotic pressure through the enzyme back into the matrix of the mitochondrion.

Oxygen is the final electron acceptor. The low-energy electrons that emerge from the electron transport system are taken up by O2. The negatively charged oxygen molecules take up protons from the medium and form water ($$\mathrm{2H^+ + 2e^- + \dfrac{1}{2} O_2 \rightarrow H_2O}$$).

### Summary of Glycolysis and Cellular Respiration

##### Glycolysis

During glycolysis, glucose (C6) is converted to two pyruvates (C3).

$\textrm{C-C-C-C-C-C} \rightarrow \textrm{C-C-C + C-C-C}$

##### Formation of Acetyl CoA

One acetyl CoA is formed for each pyruvate produced by glycolysis (see the step above).

$\textrm{C-C-C} \rightarrow \textrm{C-C} + \mathrm{CO_2}$

$\mathrm{pyruvate \rightarrow acetyl\: CoA + CO_2}$

##### Citric Acid Cycle

$\textrm{C-C} \rightarrow \mathrm{2 CO_2}$

##### Electron Transport System

In the electron transport system, NADH and FADH2 are oxidized and the energy is used to produce ATP.

### Total ATP yield per glucose

#### Conversions

NADH produced in the cytoplasm produces two to three ATP by the electron transport system.

NADH produced in the mitochondria produces approximately three ATP.

FADH2 adds its electrons to the electron transport system at a lower level than NADH, so it produces approximately two ATP.

#### Glycolysis

2 ATP

2 NADH (= 4 ATP; these are converted to ATP in the mitochondria during cellular respiration)

2 ATP

#### Total Yield

Glycolysis produces 2 ATP; aerobic respiration produces 34 more ATP

 Pathway Substrate-Level Phosphorylation Oxidative Phosphorylation Total ATP Glycolysis 2 ATP 2 NADH = 4 - 6 ATP 6 - 8 CoA 2 NADH = 6 ATP 6 Citric Acid Cycle 2 ATP 6 NADH = 18 ATP 2 FADH2 = 4 ATP 24 TOTAL 4 ATP 32 ATP 36 - 38

### Fermentation

Without oxygen, cellular respiration could not occur because oxygen serves as the final electron acceptor in the electron transport system. The electron transport system would therefore not be available.

Glycolysis can occur without oxygen. Although glycolysis does not require oxygen, it does require NAD+. Cells without oxygen available need to regenerate NAD+ from NADH so that in the absence of oxygen, at least some ATP can be made by glycolysis.

The total ATP yield of fermentation comes from glycolysis; 2 ATP molecules are produced per glucose.

#### Usefulness of Fermentation

##### Anaerobic exercise

During vigorous exercise, oxygen is consumed faster than it is needed. Additional ATP energy is provided to the muscles by glycolysis and the result is a buildup of lactate in the muscles.

When lactate builds up, the blood pH drops and the muscles fatigue.

At rest, lactate is converted back to pyruvate (the oxygen debt is repaid). This is why you continue to breathe hard after you have finished running or rapid stair climbing.

##### Yeast

Yeast produce alcohol which accumulates in their environment. As the concentration of alcohol in their environment increases, it becomes more and more toxic to them. Beer and wine have a maximum alcohol concentration because a higher concentration will kill the yeast cells.

#### Evolution of Cellular Respiration

Early cells probably fermented organic molecules in the oceans.

Today, nearly all organisms show some form of fermentation which indicates that it evolved early in evolutionary history.

Evolution typically operates by building upon or adding to what is already there. Aerobic respiration appears to have been added to fermentation.

### Summary

#### Glycolysis

Two ATP molecules are used to phosphorylate and activate glucose.

Four ATP molecules are produced by substrate-level phosphorylation.

The net yield of ATP is two; two are used and four are produced.

#### Fermentation

Fermentation is needed to regenerate NAD+ from NADH so that at least some ATP can be made in glycolysis.

Electrons from NADH are added to pyruvate (reduction) to produce alcohol (plants, yeast) or lactate (animals, bacteria)

#### Aerobic Respiration

Aerobic respiration occurs when oxygen is available.

$\mathrm{pyruvate \rightarrow CO_2 + H_2O}$

It occurs in the mitochondrion.

In the electron transport system, NADH and FADH2 are used to produce ATP as electrons are passed from one carrier to another. Eventually the electrons combine with hydrogen ions and oxygen (reduction) to form water.

Click the icon (left) to see an enlarged, color version of the diagram above.

### Review Exercise

Draw each of the following. Draw only the carbon skeletons. Do not name the molecules. Draw carbon dioxide as CO2.

• glycolysis
• formation of acetyl CoA
• citric acid cycle

• glucose
• pyruvate
• acetyl CoA