Module 3C: CELLULAR RESPIRATION

Photosynthesis shows how autotrophs (e.g. plants) convert CO2 and H2O in the presence of
sunlight to sugars, such as glucose. In glucose, energy is stored in its chemical bonds. How do plants and
animals utilize such energy to sustain life? This is accomplished through a series of metabolic pathways,
collectively called cellular respiration. Cellular respiration extracts the energy from the bonds in glucose
and converts it into a form, such as ATP(Adenosine Triphosphate), that all living things can use.

Learning Objectives:
1. To describe the metabolic pathway of cellular respiration;
2. To account the number of ATP produced in each stage of cellular respiration; and
3. To explain the process of fermentation (an anaerobic pathway of respiration).

Cellular respiration is a catabolic process involving a series of oxidation-reduction reactions; each

reaction is catalyzed by a specific enzyme. Oxidation is the loss of electrons from one substance while
reduction is the addition of electrons to another substance. However, electrons are always accompanied
by hydrogen (H+). It can occur in the presence of oxygen (aerobic respiration) or even without oxygen
(anaerobic respiration, such as fermentation).

Carbohydrates, fats, and proteins can be used as energy sources; however, it is glucose that is
most often used by cells. In cellular respiration, the significant event is the release of energy with
concomitant production of CO2 and H2O. Write the general equation for cellular respiration.

Cellular respiration involves 3 stages: glycolysis, Krebs cycle (also known as citric acid cycle), and
electron transport chain (ETC). Glycolysis is anaerobic and occurs in the cytoplasm; the Krebs cycle and
ETC are aerobic and occur in the mitochondrion (Fig. 1).

Figure 1. Overall pathway of cellular respiration.

Stage 1: Glycolysis (Fig.2)

In glycolysis, glucose (a 6C sugar) is split into 2 molecules of pyruvate (pyruvic acid). During the
process, some ATP is directly produced when phosphate groups are transferred to ADP (adenosine
diphosphate). Glycolysis involves 2 phases: energy investment and energy payoff.
 A. Energy Investment Phase
1. In the cytoplasm, a glucose molecule is phosphorylated by the transfer of a phosphate group from
ATP to form glucose-6-phosphate.
2. Glucose-6-phosphate is rearranged and converted to its isomer, fructose-6-phospate.
3. Another ATP molecule is invested wherein the phosphate group is attached to fructose-6-
phosphate resulting to fructose-1,6-bisphosphate.
4. Fructose-1,6-bisphosphate is cleaved into two 3C sugars: dihydroxyacetone phosphate (DHAP)
and glyceraldehyde-3-phosphate (G3P). These are isomers of each other and are reversibly
convertible.
5. Since only G3P is used, DHAP is converted to another G3P. Hence, from 1 glucose molecule, 2
molecules of G3P are formed.

Figure 2. The energy investment phase of glycolysis

B. Energy Payoff Phase

6. G3P is oxidized. Electrons and H+ are transferred to NAD (Nicotinamide Adenine Dinucleotide) to
form NADH. Also, a phosphate group is attached to G3P yielding 1,3-bisphosphoglycerate (BPG).
7. From BPG, a phosphate group is removed and accepted by ADP forming ATP (by substrate level
phosphorylation). The product of this reaction is 3-phosphoglycerate (3PG).
8. 3PG undergoes molecular rearrangement and 2-phosphoglycerate results.
9. From 2-phosphoglycerate, a water molecule is removed to yield phosphoenolpyruvate (PEP).
10. The phosphate group from PEP is transferred to ADP forming ATP. The resulting molecule from
this reaction is pyruvate.

Figure 3. The energy payoff phase of glycolysis.

Question: Overall, what are the products of glycolysis?

Pyruvate Oxidation
Only a small amount of chemical energy stored in glucose is released during glycolysis; most of the
energy are still found in pyruvate. If oxygen is present, pyruvate undergoes oxidation in the mitochondrial
matrix of eukaryotic cells (Fig. 4) or in the cytoplasm of prokaryotic cells. During this process, the following
events happen (Fig. 5):

1. Pyruvate undergoes decarboxylation; the carboxyl group (-COO-) is removed and released as
CO2.
2. The remaining 2C molecule (acetate) is oxidized; the electrons are transferred to NAD+ to form
NADH.
3. Coenzyme A is attached to the acetate to form acetyl coenzyme A (acetyl-CoA). Acetyl-CoA
enters the Krebs cycle.

Figure 4. The fate of pyruvate

 Figure 5. Pyruvate oxidation

Question: Overall, what are the products of pyruvate oxidation?

Stage 2: Krebs Cycle or Citric Acid Cycle (Fig. 6)

The Krebs Cycle generates more energy by further oxidizing the acetate to yield CO2 and the electrons
are transferred to electron carriers, NAD and FAD (Flavin Adenine Dinucleotide). Electron transport then
converts the energy in NADH and FADH2 to ATP.
1. The acetyl group of acetyl-CoA combines with oxaloacetate forming citrate.
2. Citrate removes one water molecule and accepts another water molecule to form its isomer
isocitrate.
3. Isocitrate undergoes oxidative decarboxylation; the electrons are accepted by NAD to produce
NADH and a molecule of CO2 is released. The product is α-ketoglutarate.
4. Another oxidative decarboxylation event happens; a CO2 molecule is lost and the resulting
compound is oxidized (NAD is reduced to NADH). At the same time, coenzyme A is attached
forming succinyl CoA.
5. Succinate is formed when CoA is displaced from succinyl CoA by a phosphate group, which is
utilized by guanosine diphosphate (GDP) to form guanosine-5’-triphosphate (GTP). GTP then
transfers the phosphate group to ADP producing ATP.
6. Succinate is oxidized; electrons are accepted by FAD to form FADH2. Fumarate is produced.
7. A water molecule is added and fumarate is converted to malate.
8. Malate is oxidized; NAD is reduced to NADH; oxaloacetate is regenerated to start a new cycle.
 Figure 6. The Krebs Cycle (or Citric Acid Cycle)

Question: Overall, what are the products of the Krebs Cycle?

Stage 3: Electron Transport Chain (ETC)and Chemiosmosis (Fig. 7)

A. Electron Transport Chain

The ETC is a series of electron carriers embedded in the inner mitochondrial membrane. NADH
and FADH2 transfer their electrons to the ETC to generate ATP by oxidative phosphorylation. The
following are the events involved:
1. Electrons from NADH are transferred to flavoprotein, the first molecule of the ETC.
2. Flavoprotein is oxidized as it passes electrons to an iron-sulfur protein (Fe.S).
3. Iron-sulfur protein passes the electrons to ubiquinone (Q).
4. FADH2 transfers electrons to ubiquinone.
5. Electrons then pass through a series of cytochromes. The last cytochrome in the chain (Cyt a3)
transfers the electrons to oxygen (the final electron acceptor). Oxygen then combines with
hydrogen ions (H+) and water is formed.

B. Chemiosmosis
The ETC does not produce ATP directly; ATP is generated because of the creation of
electrochemical proton gradient in the mitochondrion. As electrons are transported along the ETC,
protons (H+) from the mitochondrial matrix are pumped to the intermembrane space. Eventually, H+
build up in the intermembrane space creating a concentration gradient of H+. H+ then tend to move into
the mitochondrial matrix through the ATP synthase, which converts the energy of H+ flow into chemical
energy that is transferred to ATP.
 For every NADH participating in the ETC, 3 ATP are generated whereas 2 ATP are generated for
every FADH2. How much ATP then can a cell produce from 1 glucose molecule? In glycolysis, a net of
2 ATP are produced; in the Krebs cycle, another 2 ATP are synthesized; whereas the ATP synthase
apparatus generates 34 ATP. The total is 38 ATP. In eukaryotic cells, only 36 ATP are formed (Fig. 8).
Why? (For update on ATP generation, please read the link below)
https://iubmb.onlinelibrary.wiley.com/doi/abs/10.1002/bmb.2005.49403306416

Figure 7. The Electron Transport Chain and Chemiosmosis

 Figure 8. Energy (ATP) tally

Anaerobic Pathway
What happens if there is no oxygen or the amount of oxygen is insufficient after pyruvate is formed?
Another pathway exists known as anaerobic pathway. This includes fermentation. In this pathway, pyruvate
serves as the electron acceptor producing molecules (ethanol or lactic acid) that the cell cannot use.

Lactic Acid Fermentation

The fermentation method used by animals and certain bacteria, like those in yogurt, is lactic acid
fermentation (Figure 9). This type of fermentation is used routinely in mammalian red blood cells and in
skeletal muscle that has an insufficient oxygen supply to allow aerobic respiration to continue (that is, in
muscles used to the point of fatigue). In muscles, lactic acid accumulation must be removed by the blood
circulation and the lactate brought to the liver for further metabolism. The chemical reactions of lactic acid
fermentation are the following:

Pyruvic acid + NADH ↔ lactic acid + NAD+

The enzyme used in this reaction is lactate dehydrogenase (LDH). The reaction can proceed in
either direction, but the reaction from left to right is inhibited by acidic conditions. Such lactic acid
accumulation was once believed to cause muscle stiffness, fatigue, and soreness, although more recent
research disputes this hypothesis. Once the lactic acid has been removed from the muscle and circulated
to the liver, it can be reconverted into pyruvic acid and further catabolized for energy.
During lactic acid fermentation, two molecules of ATP are produced for every molecule of glucose
used (Fig. 9).

Glucose → 2 Lactate + 2 ATP

Figure 9. Fermentation in yeast and in muscle cells.

Alcohol Fermentation
Another familiar fermentation process is alcohol fermentation (Figures 9 and 10) that produces
ethanol, an alcohol. The first chemical reaction of alcohol fermentation is the following (CO2 does not
participate in the second reaction):

Pyruvic acid → CO2 + acetaldehyde + NADH → ethanol + NAD+

The first reaction is catalyzed by pyruvate decarboxylase, a cytoplasmic enzyme, with a coenzyme
of thiamine pyrophosphate (TPP, derived from vitamin B1 and also called thiamine). A carboxyl group is
removed from pyruvic acid, releasing carbon dioxide as a gas. The loss of carbon dioxide reduces the size
of the molecule by one carbon, making acetaldehyde. The second reaction is catalyzed by alcohol
dehydrogenase to oxidize NADH to NAD+ and reduce acetaldehyde to ethanol. The fermentation of pyruvic
acid by yeast produces the ethanol found in alcoholic beverages. Ethanol tolerance of yeast is variable,
ranging from about 5 percent to 21 percent, depending on the yeast strain and environmental conditions.
During alcohol fermentation (which occurs in yeasts), two molecules of ATP are produced for every
molecule of glucose used (Fig. 9).

Glucose → 2 Ethanol + 2 CO2 + 2ATP

Figure 10. Fermentation of grape juice into wine produces CO2 as a byproduct. Fermentation tanks have
valves so that the pressure inside the tanks created by the carbon dioxide produced can be released.

Other Types of Fermentation

Other fermentation methods occur in bacteria. Many prokaryotes are facultatively anaerobic. This
means that they can switch between aerobic respiration and fermentation, depending on the availability of
oxygen. Certain prokaryotes, like Clostridia, are obligate anaerobes. Obligate anaerobes live and grow in
the absence of molecular oxygen. Oxygen is a poison to these microorganisms and kills them on exposure.
It should be noted that all forms of fermentation, except lactic acid fermentation, produce gas. The
production of particular types of gas is used as an indicator of the fermentation of specific carbohydrates,
which plays a role in the laboratory identification of the bacteria. Various methods of fermentation are used
by assorted organisms to ensure an adequate supply of NAD+ for the sixth step in glycolysis. Without these
pathways, that step would not occur and no ATP would be harvested from the breakdown of glucose.