Chapter 7: How cells harvest energy

Autotrophs & Heterotrophs

Autotrophs Heterotrophs
(Self-feeders) (Fed by others)
Mechanism Absorbing radiant energy of Feed on the organic
sunlight and use it to form compounds produced by
organic compounds through autotrophs
photosynthesis
Examples Plants, algae and some All animals and fungi, and
bacteria most protists and
prokaryotes

Extraction of Potential Energy by Heterotrophs

 Potential energy is stored in C-H bonds of foods which contains energy-rich organic
compounds such as carbohydrates, proteins and fats
 Chemical reaction which harvests the potential energy from chemical bonds are
oxidation reactions
 Cells utilize enzyme-facilitated redox reactions to take energy from food sources and
convert it to ATP. This process is known as cellular respiration
Cellular respiration

Cellular respiration refers to a series of metabolic processes that take place within a cell in
which biochemical energy is harvested from organic substance (e.g. glucose) and then
stored in an energy-rich ATP for use in energy-requiring activities of the cell.

 In cellular respiration, a number of redox reactions take place, and a number of

molecules, including NAD+, act as electron carriers
 During each transfer of electrons, energy is released. A substantial portion of this energy
is captured and used to make ATP or to form other chemical bonds; the rest is lost as
heat
 The ultimate goal of cellular respiration is to produce ATP

3 main types of cellular respiration which differs in the final electron acceptor:

-O2
Anaerobic respiration
+O2 Final e- acceptor = Inorganic molecule (not O2)
Aerobic Cellular
respiration respiration Fermentation
Final e acceptor = O2
-
Final e- acceptor = Organic molecule (not O2)
Aerobic Respiration
 In aerobic respiration, the final electron acceptor is oxygen (O 2)
 Free energy released = – 686 kcal/mol of glucose (when glucose is oxidized)
 This large amount of energy must be released in small steps rather than all at once 
 Oxidation of glucose by aerobic respiration also results in the release of 6 carbon dioxide
and 6 water molecules 
 C6H12O6 + 6O2 → 6CO2 + 6H2O + ATP

Electron Carriers: NAD+ and NADH

Electron Carriers
 Molecules that transfer electrons from one molecule to another
 Some only carries electrons, some carries both electrons and proton
 Can be reversibly oxidized and reduced
 Aid in the gradual, stepwise release of the energy from oxidation
 Can exist as:
 Soluble carrier e.g. NAD+ and FAD2+
 Membrane bound carriers that form a redox chain 
 Carrier that moves within the membrane

Structure of NAD+ (nicotinamide adenosine dinucleotide)

 Composed of 2 two nucleotides joined head-to-head by their phosphate groups
 The two nucleotides are:

1. Nicotinamide monophosphate (NMP)

 The active part of the molecule
 Easily accepts electrons and being reduced

2. Adenosine monophosphate (AMP)

 Act as the core
 Providing a shape recognized by many enzymes

NAD+ and NADH: This dinucleotide serves as an “electron shuttle” during cellular

respiration. NAD+ accepts a pair of electrons and a proton from catabolized macromolecules
and is reduced to NADH. NADH now carries the energetic electrons and can supply them to
other molecules and reduce them. It also acts as a cofactor in oxidation-reduction reactions.
Two electrons and a proton are transferred to NAD+ with another proton donated to the
solution. Molecules that gain electrons are said to be reduced, and ones that lose energetic
electrons are said to be oxidized. NAD+ oxidizes energy-rich molecules by acquiring their
electrons
FAD2+ and FADH
 FAD2+ = flavin adenine dinucleotide
** we refer to electron carriers as FAD2+ not FADH

Substrate Level Phosphorylation & Oxidative Phosphorylation

Synthesis of ATP
 There are 2 ways by which ATP can be synthesized in the cell:
1. Substrate-level phosphorylation
2. Oxidative phosphorylation

Substrate-level Phosphorylation
 ATP is formed by transferring a phosphate group directly to ADP from a phosphate-
bearing molecule
 For example: phosphoenolpyruvate (PEP) – has a high-energy phosphate bond similar to
the bonds in ATP
 PEP’s phosphate group is transferred enzymatically to ADP to create ATP
 Produces 4 ATP in aerobic respiration
Aerobic respiration
1. Glycolysis (cytoplasm)
2. Pyruvate oxidation (mito matrix)
3. Krebs Cycle (mito matrix)
4. Electron Transport Chain/ Chemiosmosis (mito. Cristae)

Oxidative Phosphorylation
 ATP is synthesized from ADP by the enzyme ATP synthase, using energy from a proton
(H+) gradient, which is generated by transfer of high-energy electrons (from glucose
oxidation) ferried by NADH and FADH2 to O2 via a series of electron carriers (in the ETC)
 The energy from the proton gradient is used to catalyse the reaction : ADP + Pi → ATP
 Produces 28 ATP in aerobic respiration

Aerobic respiration
5. Glycolysis (cytoplasm)
6. Pyruvate oxidation (mito matrix)
7. Krebs Cycle (mito matrix)
8. Electron Transport Chain/ Chemiosmosis (mito. Cristae)

Substrate-level phosphorylation Oxidative phosphorylation

Formation of ATP by transferring a Formation of ATP by the enzyme, ATP
phosphate group directly to ADP from a synthase, using energy from a proton (H+)
phosphate-bearing molecule gradient
 1 glucose molecule

It takes 1ATP to transport

1. Net yield = 2 pyruvate + 2 NADH + 2 ATP each molecule of NADH
that is generated by
glycolysis in the
2. Net yield = 2 Acetyl-CoA+ 2 CO2 + 2 NADH cytoplasm into the
mitochondria *

3. Net yield = 4 CO2 + 6 NADH + 2 FADH2 + 2 ATP

4. 10 NADH 2 FADH2 4 ATP

x2.5 ATP x1.5 ATP ATP produced /
30
glucose molecule
25 ATP 3 ATP 2* ATP
Stage 1: Glycolysis

 Converts glucose into two 3-carbon molecules of pyruvates

 Occurs in the cytoplasm
 Also results in net gain of 2 ATP molecules and 2 NADH
 Involves 10 reactions:
 Reactions 1-3: Priming reactions - converts glucose to fructose 1,6-biphosphate
(requires 2 ATP)
 Reactions 4-5: Cleavage - results in 2 Glyceraldehyde 3-phosphate (G3P) molecules
 Reactions 6-10: Oxidation and ATP formation - results in 2 pyruvate molecules
(produce 2 NADH and 4 ATP)

Rxn Activity Results

1-3 Priming Glucose  Fructose 1,6 biphosphate
reactions (Impt: requires 2 ATP )
4-5 Cleavage 2 Glyceraldehyde-3-phosphate (G3P) molecules
(lysis)
6-10 Oxidation & 2 Pyruvate molecules (+ 2 NADH + 4 ATP are
ATP produced)
formation

The 10 reactions of glycolysis. The first half of glycolysis consists of five sequential reactions
that convert one molecule of glucose into two molecules of the 3-carbon compound
glyceraldehyde 3-phosphate (G3P). These reactions require the expenditure of ATP, so they
constitute an endergonic process. In the second half of glycolysis, five more reactions
convert G3P into pyruvates in an energy-yielding process that generates ATP. This 10-step
biochemical pathway occurs in the cytoplasm where it converts 1 glucose (6 carbons) to 2
pyruvate (3 carbons)

Net yield (per glucose molecule): 2 Pyruvate + 2 NADH + 2 ATP

Stage 2: Pyruvate oxidation

 In the presence of oxygen, pyruvate is oxidized to acetyl-CoA

 Occurs in the mitochondria matrix in eukaryotes
 Multienzyme complex called pyruvate dehydrogenase catalyzes the reaction

Products of Pyruvate Oxidation

 Each pyruvate molecule will produce: 1 CO2 + 1 NADH + 1 Acetyl-CoA
 Each glucose molecule will produce: 2 CO2 + 2 NADH + 2 Acetyl-CoA
 AKA 2 pyruvate  2 acetyl CoA + 2 CO2 + 2 NADH
(Since one glucose molecule is converted into 2 pyruvate molecules during glycolysis.)
Stage 3: Krebs cycle

 Oxidizes the acetyl group from pyruvate

 Occurs in the matrix of the mitochondria
 Biochemical pathway of 9 steps in 3 segments

1. Acetyl-CoA + oxaloacetate → citrate

2. Citrate rearrangement and decarboxylation to form a new 4-carbon molecule
succinate
3. Regeneration of oxaloacetate

Step of the Krebs cycle:

Step 1. In the first step of the citric acid cycle, acetyl CoA joins with a four-carbon molecule,
oxaloacetate, releasing the CoA group and forming a six-carbon molecule called citrate.
Step 2 to 5. The citrate is converted to a 5-carbon intermediate (α-ketoglutarate), and then
converted into 4-carbon succinate. Oxidation reactions combined with 2 decarboxylation
reactions, 2 CO2, 2 NADH and 1 ATP are produced.
Step 6 to 9. The succinate undergoes three additional reactions to regenerate the 4C
oxaloacetate again. During these reactions 1 NADH is produced, a molecule of flavin adenine
dinucleotide (FAD), another cofactor, becomes reduced to 1 FADH2.

Summary
1. Reaction 1: Condensation
2-3. Reaction 2 & 3: Isomerization
4. Reaction 4: The first oxidation
5. Reaction 5: The second oxidation
6. Reaction 6: Substrate-level phosphorylation
7. Reaction 7: The third oxidation
8-9. Reaction 8 & 9: Regeneration of oxaloacetate and the fourth oxidation
Products of the Krebs Cycle

For each Acetyl-CoA entering the Krebs Cycle:

 2 molecules of CO2 are released

 3 NAD+ are reduced to 3 NADH
 1 FAD (electron carrier) is reduced to 1 FADH2  
 1 ATP is produced
 Oxaloacetate is regenerated

Total products of glycolysis + pyruvate oxidation + Krebs cycle

One glucose molecule has been oxidized to:

 6 CO2
 4 ATP
 10 NADH
 2 FADH2
Stage 4: Electron Transport Chain & Chemosis

4 4
2

Electron Transport Chain

** A complex is a structure that consists of a central atom/ molecule/ protein that is

weakly-connected to surrounding atoms/molecules/proteins.
** Cytochromes are a group of proteins that have Fe atoms as cofactors

 The NADH and FADH2 carry energetic electrons to the electron transport chain
 ETC is a series of electron carriers embedded in the inner mitochondrial membrane
 NADH will transfer the 2 electrons to NADH dehydrogenase in the ETC
 FADH2 will transfer the 2 electrons to ubiquinone in the ETC
 The electrons are transferred to the complexes NADH dehydrogenase, bc1 complex &
cytochrome oxidase complex
 Mobile carriers ubiquinone transfers electrons from NADH dehydrogenase to bc1
complex and Cytochrome c transfers electrons from bc1 complex to cytochrome oxidase
complex
 Each complex uses a portion of the electron’s energy to pump protons out of the
mitochondrial matrix into the intermembrane space, creating a proton gradient
 NADH dehydrogenase and bc1 complex transport 4 protons, and the cytochrome
oxidase complex transports 2 protons
 The electrons are finally used to reduce oxygen, forming water.
 Protons return to the matrix through the membrane by diffusion
 Complex/ Event No. of H+ No. of H+
molecule pumped out pumped out
from 1 NADH: from 1 FADH2:
1/NADH Redox of NADH + H+  4H+ 0
dehydrogenase NAD+, 2e- move to Q
Q Redox of FADH2  FAD 0 0
2/ bc1 complex Carrier C (Cytochrome 4H+ 4H+
C) transports 2e- to
complex IV
3/ Cytochrome O2 (in matrix) accepts 2H+ 2H+
oxidase complex the 2e-; leading to
formation of H2O
O2 + 4H+ + 4e-  2H2O
Total 10H+ 6H+
4H+ are required to make 1 ATP 2.5 ATP 1.5 ATP
Number of H+ pumped out by 1 glucose 10H+ x 10 = 6H x 2 = 12H+
+

molecule 100 H+

Chemiosmosis
Definition: Chemiosmosis is the movement of ions across a selectively permeable
membrane, down their concentration gradient.
 Uses a proton gradient to make ATP
 Accumulation of protons in the intermembrane space drives protons into the matrix via
diffusion
 Membrane is relatively impermeable to ions
 Most protons will re-enter the matrix through ATP synthase which is a transmembrane
proton channel protein
 The movement of proton down its concentration gradient provide the energy for the
synthesis of ATP from ADP + Pi
Yield of ATP
 The number of ATP molecules produced by ATP synthase per molecule of glucose
depends on:

1. The number of protons transported across the inner membrane (in the electron
transport chain)
 One NADH can transport 10 H+
 One FADH2 can transport 6 H+

2. The number of protons needed per ATP synthesized (when passing through the ATP
synthase)
 4 H+ are required to synthesize 1 ATP
 Therefore:

o Each NADH is able to produce 2.5 ATP

o Each FADH2 is able to produce 1.5 ATP

Theoretical ATP yield. The theoretical yield of ATP harvested from glucose by aerobic
respiration totals 32 molecules (28 of them are synthesized by oxidative
phosphorylation and 4 of them by substrate-level phosphorylation). In eukaryotes
this is reduced to 30 because it takes 1 ATP to transport each molecule of NADH that
is generated by glycolysis in the cytoplasm into the mitochondria. 

 32 ATP per G in prokaryotes

 30 ATP per G in eukaryotes due to the transport of cytoplasmic NADH into mitochondria
 Yield

Stage Site of Initial substrate Intermediate Final product Net ATP NADH FADH2 CO2
occurrence molecule/s
Glycolysis Cytoplasm One glucose 2 Pyruvate 2 2 0 0
molecule
2 Glyceraldehyde 3-
phosphate
&
1 fructose 1,6-
biphosphate

Pyruvate Mitochondria 2 Pyruvate 2 Acetyl-CoA 0 2 0 2

Oxidation

NIl

Krebs cycle Mitochondria 2 Acetyl-CoA Two citrate 2 Oxaloacetate 2 6 2 4

Total 4 10 2 6

x2.5 ATP x1.5 ATP

2* ATP 25 ATP 3 ATP

 Regulation of Aerobic Respiration

 Regulation of aerobic respiration is by feedback inhibition at two key control points:

1. In glycolysis: Phosphofructokinase is allosterically inhibited by ATP and/or citrate
2. In pyruvate oxidation: Pyruvate dehydrogenase inhibited by high levels of NADH

Allosteric Source of Inhibitors The stage The enzyme The reaction inhibited Allosteric activators
Inhibitors inhibited/ inhibited
activated

Glycolysis, Krebs cycle, Glycolysis and Phosphofructokinase Conversion of fructose 6- ADP

ATP Chemiosmosis thus an phosphate to fructose 1,6-
decreased/ biphosphate
increased level
of pyruvate
Krebs cycle Glycolysis and Phosphofructokinase Conversion of fructose 6-
Citrate thus an phosphate to fructose 1,6-
decreased/ biphosphate
increased level
of pyruvate
Krebs cycle pyruvate Pyruvate Conversion of pyruvate to Nil
NADH oxidation and dehydrogenase Acetyl-CoA
thus a decrease
level of acetyl-
CoA
Anaerobic Respiration & Fermentation

Anaerobic Respiration
 Anaerobic respiration use inorganic molecules as the final electron acceptor such as
Inorganic Sulfur, iron, nitrate, sulphate (Due to no C or O inside them)
 E.g. Sulfur bacteria: use inorganic sulfate (SO4) as the final electron acceptor and
reduced it to hydrogen sulfide (H2S)
 Thermoproteus tenax. This organism can use elemental Sulfur as a final electron
acceptor for anaerobic respiration. 
 Thermoproteus is often found in Sulfur-containing hot springs

Fermentation
 Use organic molecules as the final electron acceptor
 e.g. 2-Acetaldehyde in ethanol fermentation by yeast
 e.g. Pyruvate in lactic acid fermentation by animal cell (muscle cells)
 Under these conditions, the electrons generated by glycolysis are donated to organic
molecules (other than Oxygen) in a process called fermentation.
 This process recycles NAD+, the electron acceptor that allows glycolysis to proceed

 In ethanol fermentation by yeast: Electrons are transferred from NADH to 2-

Acetaldehyde (derived from pyruvate) to produce ethanol
 In lactic acid fermentation in muscle cells: Electrons are transferred from NADH
to pyruvate to produce lactic acid
 Hence the final e- acceptors are Acetaldehyde and pyruvate receiving e- and H+ from
NADH
Fate of Pyruvate
Presence of O2 Absence of O2
Pyruvate molecule will be oxidised to Pyruvate is reduced to Latic acid, which is
from Acetyl-CoA, which will then enter coupled with the oxidation of NADH to
the Krebs cycle regenerate NAD+. This process is called
fermentation
Catabolism of Proteins & Fats

Catabolism of Protein
 Proteins are first broken down into their individual amino acid
 Amino acids then undergo deamination to remove the amino group
 Remainder of the amino acid is converted to a molecule that enters glycolysis or the
Krebs cycle and contribute to ATP formation e.g.:
 Alanine is converted to pyruvate
 Aspartate is converted to oxaloacetate
 Glutamate is converted to α-ketoglutarate

Catabolism of Fat

 Fats are first broken down to fatty acids and glycerol

 Fatty acids are then converted to acetyl-CoA by β-oxidation 
 The acetyl-CoA  produced will enter the Krebs cycle and contribute to ATP formation

β oxidation. Through a series of reactions known as β oxidation, the last two carbons in a

fatty acid combine with coenzyme A to form acetyl-CoA, which enters the Krebs cycle. The
fatty acid, now two carbons shorter, enters the pathway again and keeps re-entering until
all its carbons have been used to form acetyl-CoA molecules. Each round of β oxidation uses
one molecule of ATP and generates one molecule each of FADH2 and NADH.