BIO 202 — Cell Biology

Lecture 9: Cellular Respiration & Energy Metabolism

Date: Thursday, 14 March 2024
Professor: Dr. Adaeze Nwosu | Room: SCI-214
Student: [Your Name] | Student #: [Your ID]
Department of Biological Sciences, Semester 1 — 2024

1. Overview & Learning Objectives

This lecture covers the complete process of cellular respiration — the metabolic pathways by which
living cells break down glucose and other organic molecules to produce ATP (adenosine triphosphate),
the universal energy currency of the cell. Understanding these processes is fundamental to cell biology,
biochemistry, medicine, and physiology.
By the end of this lecture, students should be able to:
• Describe the three main stages of cellular respiration: glycolysis, the Krebs cycle, and the
electron transport chain.
• Explain where each stage occurs within the cell and under what conditions (aerobic vs.
anaerobic).
• Calculate the net ATP yield from the complete oxidation of one glucose molecule.
• Understand the roles of electron carriers NADH and FADH2 in energy transfer.
• Distinguish between substrate-level phosphorylation and oxidative phosphorylation.
• Explain the chemiosmotic theory and the role of ATP synthase.
• Describe the processes of fermentation as an anaerobic alternative.
• Connect cellular respiration to other metabolic pathways including glycogenolysis and beta-
oxidation.

Overall reaction summary:

C6H12O6 + 6O2 --> 6CO2 + 6H2O + ~36-38 ATP

2. Background: Why Cells Need Energy

All living organisms require a continuous supply of energy to maintain their structure, carry out
chemical reactions, grow, reproduce, and respond to their environment. In eukaryotic cells, this energy
primarily comes from the oxidation of organic molecules — mainly glucose — through the process of
cellular respiration.
ATP is the molecule that directly powers most cellular work. When the terminal phosphate bond of
ATP is hydrolyzed (split), energy is released that can drive endergonic (energy-requiring) reactions. The
cell constantly regenerates ATP from ADP and inorganic phosphate (Pi) using the energy stored in food
molecules.
2.1 Redox Reactions in Respiration
Cellular respiration is fundamentally a series of oxidation-reduction (redox) reactions. Glucose is
progressively oxidized (loses electrons/hydrogen atoms), while oxygen is ultimately reduced (gains
electrons) to form water. The electrons do not move directly from glucose to oxygen; instead, they are
passed along a chain of electron carriers.
Key electron carriers:
• NAD+ (nicotinamide adenine dinucleotide) — accepts 2 electrons + 1 H+ to become NADH
• FAD (flavin adenine dinucleotide) — accepts 2 electrons + 2 H+ to become FADH2
• Both NADH and FADH2 deliver electrons to the electron transport chain

3. Stage 1: Glycolysis
Location: Cytoplasm (cytosol). Does NOT require mitochondria. Can occur under both aerobic and
anaerobic conditions.
Glycolysis (from Greek: glykys = sweet, lysis = splitting) is the splitting of glucose (6-carbon) into two
molecules of pyruvate (3-carbon). It is an ancient metabolic pathway present in virtually all living
organisms.

3.1 Two Phases of Glycolysis

Phase 1: Energy Investment (Steps 1-5)
The cell spends 2 ATP to phosphorylate glucose and convert it to glyceraldehyde-3-phosphate (G3P).
This destabilizes the glucose molecule and prepares it for the energy-releasing phase.
1. Step 1: Glucose + ATP --> Glucose-6-phosphate (enzyme: hexokinase)
2. Step 2: Glucose-6-phosphate --> Fructose-6-phosphate (isomerization)
3. Step 3: Fructose-6-phosphate + ATP --> Fructose-1,6-bisphosphate (enzyme:
phosphofructokinase — key regulatory step!)
4. Steps 4-5: 6C molecule split into two 3C molecules (G3P)
Phase 2: Energy Payoff (Steps 6-10)
Each of the two G3P molecules is converted to pyruvate, generating ATP and NADH in the process.
Because there are two G3P molecules per original glucose, all yields below are per glucose.
5. Step 6: G3P oxidized --> 1,3-bisphosphoglycerate; 2 NAD+ reduced to 2 NADH
6. Steps 7-8: 2 ATP produced by substrate-level phosphorylation (x2 = 4 ATP total)
7. Steps 9-10: 2 more ATP produced; pyruvate formed

3.2 Net Products of Glycolysis (per glucose)

• 2 ATP net (4 produced - 2 invested)
• 2 NADH
• 2 pyruvate (3-carbon molecules)
• 2 H2O
EXAM TIP: Remember glycolysis USES 2 ATP before gaining 4. Net gain = 2 ATP. This is one of the most
common mistakes students make — confusing gross (4) vs. net (2) ATP yield from glycolysis.
3.3 Regulation of Glycolysis
Glycolysis is tightly regulated to match the cell's energy needs. The key regulatory enzyme is
phosphofructokinase (PFK), which catalyzes Step 3. PFK is:
• Inhibited by high ATP levels (cell has enough energy — slow down)
• Inhibited by high citrate levels (Krebs cycle intermediates building up)
• Activated by high AMP/ADP levels (cell needs more energy — speed up)
• Activated by fructose-2,6-bisphosphate (hormonal signal to increase glycolysis)

4. Bridge Step: Pyruvate Oxidation

Location: Mitochondrial matrix (inner compartment). Pyruvate produced in the cytoplasm is actively
transported across the inner mitochondrial membrane into the matrix.
Each pyruvate is converted into Acetyl-CoA (2-carbon) by the pyruvate dehydrogenase complex — a
large multi-enzyme assembly. This is an irreversible step.
Per pyruvate (x2 per glucose):
• 1 CO2 released (decarboxylation)
• 1 NADH produced
• 1 Acetyl-CoA formed (2-carbon unit that enters Krebs cycle)
Per glucose total from pyruvate oxidation: 2 CO2, 2 NADH, 2 Acetyl-CoA.

5. Stage 2: The Krebs Cycle (Citric Acid Cycle)

Location: Mitochondrial matrix. Also called the citric acid cycle (after its first intermediate) or the TCA
cycle (tricarboxylic acid cycle). Discovered by Hans Krebs in 1937 (Nobel Prize 1953).
The Krebs cycle is a series of 8 enzymatic reactions that completely oxidize Acetyl-CoA, extracting its
electrons for the ETC and releasing the remaining carbon as CO2. The cycle regenerates oxaloacetate
so it can accept another Acetyl-CoA and repeat.
Because one glucose produces TWO Acetyl-CoA, the cycle runs TWICE per glucose molecule.

5.1 Steps of the Krebs Cycle (per turn)

8. Acetyl-CoA (2C) + Oxaloacetate (4C) --> Citrate (6C) [enzyme: citrate synthase]
9. Citrate rearranged to Isocitrate
10. Isocitrate --> alpha-ketoglutarate (5C): CO2 released, NADH produced
11. Alpha-ketoglutarate --> Succinyl-CoA (4C): CO2 released, NADH produced
12. Succinyl-CoA --> Succinate: 1 ATP (or GTP) produced (substrate-level phosphorylation)
13. Succinate --> Fumarate: FADH2 produced
14. Fumarate --> Malate: H2O added
15. Malate --> Oxaloacetate: NADH produced; cycle is complete

5.2 Products per Krebs Cycle Turn (x2 for glucose)

• 3 NADH (x2 = 6 NADH per glucose)
 • 1 FADH2 (x2 = 2 FADH2 per glucose)
• 1 ATP (x2 = 2 ATP per glucose)
• 2 CO2 (x2 = 4 CO2 per glucose)

5.3 Regulation of the Krebs Cycle

Three key regulatory enzymes respond to energy status:
• Citrate synthase — inhibited by ATP and NADH; activated by ADP
• Isocitrate dehydrogenase — inhibited by ATP and NADH; activated by ADP and Ca2+
• Alpha-ketoglutarate dehydrogenase — inhibited by succinyl-CoA and NADH

6. Stage 3: Electron Transport Chain (ETC) & Oxidative

Phosphorylation
Location: Inner mitochondrial membrane. This is where the majority of ATP is produced —
approximately 32-34 ATP per glucose. The ETC consists of four large protein complexes (I, II, III, IV) and
two mobile electron carriers (ubiquinone/CoQ and cytochrome c).

6.1 The Four Protein Complexes

• Complex I (NADH dehydrogenase)
- Accepts electrons from NADH; pumps 4 H+ across inner membrane into intermembrane space;
passes electrons to CoQ.
• Complex II (Succinate dehydrogenase)
- Accepts electrons from FADH2; does NOT pump H+ (important! FADH2 produces less ATP than
NADH); passes electrons to CoQ.
• Complex III (Cytochrome bc1 complex)
- Accepts electrons from CoQ; pumps 4 H+ across membrane; passes electrons to cytochrome c.
• Complex IV (Cytochrome c oxidase)
- Accepts electrons from cytochrome c; pumps 2 H+ across membrane; transfers electrons to O2,
forming H2O. This is why oxygen is essential.

6.2 Chemiosmosis and ATP Synthase

As electrons move through the ETC, Complexes I, III, and IV pump H+ ions (protons) from the matrix
into the intermembrane space. This creates an electrochemical gradient — the proton-motive force
(PMF) — consisting of:
• A chemical gradient: high H+ concentration in intermembrane space vs. matrix
• An electrical gradient: intermembrane space is positively charged relative to matrix
H+ ions flow back into the matrix through ATP synthase (Complex V) — a remarkable molecular motor.
This flow of protons (chemiosmosis) drives the rotation of ATP synthase, which catalyzes the synthesis
of ATP from ADP + Pi. This process is called oxidative phosphorylation.
ATP yield from ETC: 1 NADH = ~2.5 ATP | 1 FADH2 = ~1.5 ATP
Note: These values (2.5 and 1.5) reflect current research. Older textbooks list 3 and 2 — both are acceptable in
exams unless Prof specifies.
7. Anaerobic Respiration & Fermentation
When oxygen is unavailable, cells cannot run the ETC. To continue producing ATP (via glycolysis), cells
must regenerate NAD+ from NADH — otherwise glycolysis halts. This is achieved through
fermentation.

7.1 Lactic Acid Fermentation

Occurs in: muscle cells during intense exercise, red blood cells (no mitochondria), many bacteria
(yogurt, cheese).
Pyruvate is reduced to lactate, oxidizing NADH back to NAD+. No CO2 is produced. Net yield: only 2
ATP per glucose.

7.2 Alcoholic Fermentation

Occurs in: yeast, some plant cells. Pyruvate is converted to acetaldehyde (releasing CO2), then to
ethanol. NADH is oxidized to NAD+ in the process. This is the basis of bread rising and alcohol
production.

8. Complete ATP Accounting (per glucose)

Stage-by-stage summary:
• Glycolysis: 2 ATP (net) + 2 NADH
• Pyruvate Oxidation: 0 ATP + 2 NADH
• Krebs Cycle: 2 ATP + 6 NADH + 2 FADH2
• ETC from NADH (10 total x 2.5): ~25 ATP
• ETC from FADH2 (2 total x 1.5): ~3 ATP
• TOTAL: ~30-32 ATP per glucose (theoretical maximum: ~36-38 ATP)
Why less than theoretical max? Some H+ leaks across the inner membrane without passing through
ATP synthase. Energy is also used to transport pyruvate and ADP/ATP across the membrane.

9. Key Concepts & Vocabulary

ATP (adenosine triphosphate): Energy currency of the cell. Hydrolysis of its terminal phosphate
releases ~7.3 kcal/mol.
NADH / FADH2: Reduced electron carriers. Deliver high-energy electrons to ETC for ATP synthesis.
Substrate-level phosphorylation: Direct ATP synthesis when a phosphate group is transferred from a
substrate to ADP. Occurs in glycolysis and Krebs.
Oxidative phosphorylation: ATP synthesis driven by ETC and chemiosmosis. Accounts for ~90% of total
ATP.
Chemiosmosis: ATP synthesis powered by proton gradient across a membrane. Proposed by Peter
Mitchell (Nobel Prize 1978).
Proton-motive force (PMF): The electrochemical gradient created by H+ pumping across the inner
mitochondrial membrane.
Aerobic respiration: Requires O2 as final electron acceptor. Produces ~36-38 ATP per glucose.
Anaerobic respiration: Does not use O2. Produces only 2 ATP (via glycolysis + fermentation to
regenerate NAD+).
Pyruvate dehydrogenase complex: Multi-enzyme complex that converts pyruvate to Acetyl-CoA.
Bridge between glycolysis and Krebs.
Oxaloacetate: 4-carbon Krebs cycle regenerator. Combines with Acetyl-CoA to form citrate and
restarts the cycle.

10. Common Exam Mistakes to Avoid

• Confusing gross vs. net ATP from glycolysis (4 produced - 2 invested = NET 2 ATP)
• Forgetting that the Krebs cycle runs TWICE per glucose (because glycolysis makes 2 pyruvate)
• Saying oxygen is used in glycolysis — it is NOT. Glycolysis is anaerobic.
• Confusing NADH with NAD+. NADH is the reduced, energy-carrying form.
• Placing glycolysis in the mitochondria — it occurs in the CYTOPLASM.
• Forgetting that Complex II does NOT pump protons (FADH2 enters at a lower point than NADH).
• Stating that fermentation produces ATP — it does NOT. Fermentation only regenerates NAD+
so glycolysis can continue.
• Using the old textbook values (3 ATP per NADH, 2 per FADH2) when current values (2.5 and 1.5)
are required.

11. Practice Questions

16. How many total ATP molecules are produced when 3 glucose molecules are completely
oxidized under aerobic conditions?
17. A cell is exposed to cyanide, which blocks Complex IV of the ETC. What would happen to NADH
levels? Why?
18. Compare and contrast substrate-level phosphorylation and oxidative phosphorylation.
19. Why does a runner 'feel the burn' in their muscles during a sprint? Explain in terms of cellular
respiration.
20. A mutation disables ATP synthase. How would this affect the proton gradient? What cellular
processes would be affected?
21. How does high ADP concentration affect glycolysis? What is the molecular mechanism?
22. Yeast placed in an anaerobic environment begins producing CO2. Explain which metabolic
process is responsible and why.

UPCOMING: Quiz on Cellular Respiration — Tuesday, March 19. Covers Lectures 8-9. Bring calculator.
30 minutes.
ESSAY DUE: Week 15 — 'Compare aerobic and anaerobic respiration in the context of human athletic
performance.' 1,500 words, APA format.
READING: Campbell & Reece, Biology 12th Ed., Chapter 9 (pp. 164-196). Focus on Figures 9.6, 9.12,
9.16.
LAB: Fermentation experiment with yeast — Report due Friday, March 22. Follow lab manual protocol
Section 4B.