Bioenergetics

(Energy of metabolites in cells)

 Why do we eat?
To get Energy  To do work
Mechanical work: muscle contraction

Electrical work: transmission of nerve impulses

Chemical work: building up molecules (anabolism)

e.g proteins

Osmotic work: absorption , secretion, active

transport etc.
 What do we eat?
Carbohydrates
Lipids Energy
Proteins (ATP)

Vitamins & minerals (micronutrients)

How does your body convert
what you eat into energy?
Metabolism
 Metabolism
It involves all the biochemical changes (reactions)
that various foodstuffs (Nutrients) undergo inside
the body.

Metabolic pathway: It is a series of chemical reactions

occurring within a cell.
In this series of reactions, the product of each step in
the series is the starting material for the next step.
ENZ-1 ENZ-2 ENZ-3
AA B C D

The reactants, intermediates, and products of this

series are called metabolites.
Metabolism includes catabolism & anabolism

I- Anabolism: is the process of building up of

macromolecules (using the energy created in
converting ATP to ADP) e.g. glycogen, proteins and
lipids.
Catabolism: is the process of breaking down
macromolecules into simpler compounds.

Catabolic reactions are exothermic, involving

oxidative reaction, and commonly producing reduced
equivalents (NADH and FADH). These reduced
equivalents are mainly oxidized via the respiratory
chain with production of ATP.
Catabolism is classified into three main stages:
 Different Stages of Catabolism

Carbohydrates Proteins Lipids

Stage One:
No enough energy (no ATP) released (breakdown of low energy bonds)

Monosaccharides Amino Acids Glycerol + Fatty Acids

Stage Two:
Some Energy (ATP) is produced (breakdown of high energy bonds)

Acetyl-CoA
Stage Three: (Mitochondria)
Oxidized H2O + ATP +
Most of energy (ATP) will be Coenzymes Free Energy
produced in this stage
Electron
Krebs’Cycle
Reduced
Transport Chain
Coenzymes [O] + ADP + Pi

CO2
 ATP-ADP Cycle
Def: The cycle that connects the processes that
generate & consume high-energy phosphate (~ P).

Rate: The cycle occurs at a very rapid rate since the

total ATP/ADP pool is very small and the amount of
ATP present in any cell is sufficient to maintain its
activity for only a few seconds.
Storage:
• ATP very unstable to be stored
• Creatine phosphate is the major storage form of
high-energy phosphate in muscles.
• It is formed from ATP in energy-rich states. Creatine
phosphate gives its high-energy phosphate back to
ADP to form ATP in energy-poor states (muscle
contraction).
Energy Levels of Hydrolysable Bonds
Hydrolysable bonds are divided into 2 groups:
I-Low Energy Bonds
•These are bonds that upon hydrolysis liberate an amount
of free energy less than 7.3 Kcal/mole.
•
The bond is represented by an ordinary dash (−).
 II -High Energy Bonds
•These are bonds that upon hydrolysis liberate an amount of free
energy ≥ 7.3 Kcal/mole (7.3 – 14.8).

•The bond is represented by a double curved dash (~).

A- High Energy Phosphate Bonds

OH

~ P = ~ P=O

OH
Example: Pyrophosphate bond
This is the bond between two phosphates, such as that
connecting the terminal 2 phosphates in ATP.
γ β α

O O O A

HO - P ~ O – P ~ O – P – O – CH2
O

OH OH OH

H H

` 2`
H H
ATP ADP AMP

OH OH
III-Generation of High Energy Phosphate
Bonds

A- ELECTRON TRANSPORT CHAIN (ETC)

(Respiratory Chain)
 Different Stages of Catabolism

Carbohydrates Proteins Lipids

Stage One:
No enough energy (no ATP) released (breakdown of low energy bonds)

Monosaccharides Amino Acids Glycerol + Fatty Acids

Stage Two:
Some Energy (ATP) is produced (breakdown of high energy bonds)

Acetyl-CoA
Stage Three: (Mitochondria)
Oxidized H2O + ATP +
Most of energy (ATP) will be Coenzymes Free Energy
produced in this stage
Electron
Krebs’Cycle
Reduced
Transport Chain
Coenzymes [O] + ADP + Pi

CO2
 Oxidation Reduction

Addition of oxygen Addition of hydrogen

Removal of hydrogen Removal of oxygen

Loss of electrons Gain of electrons

Redox Reactions
•The oxidation-reduction reactions are called redox reactions.

•The oxidized and reduced forms of the same substrate of a redox

reaction are known as redox pair or redox couple.

•For example, NAD (oxidized form)/NADH (reduced form)

constitutes a redox couple.
•Redox reaction occurs, when there is transfer of electrons between
an electron donor (reducing agent) and an electron acceptor
(oxidizing agent).
•Redox reactions are accompanied by release of free energy. .
.
•The amount of released energy depends on the difference
in the electric potential between the donor and the acceptor.
Electrons are transferred from redox pairs of lower
electric potential (have higher tendency to loose
electron) to redox pairs of higher electric potential
(higher tendency to gain electron).
•Electron potential: tendency to acquire electrons & hence
to be reduced
 Electron Transport Chain
(Respiratory chain)
Oxidative phosphorylation

Site: Inner Mitochondrial Membrane

Components:
• 4 integral protein complexes I,II,III and IV.
• 2 Mobile carriers
• Complex V (ATP synthase)

Each complex accepts or donates electrons to a

mobile electron carrier.
1- Complex I (NADH dehydrogenase complex)
• It is a flavoprotein bound to FMN.
• It catalyzes transfer of 2 hydrogens from NADH to
ubiquinone to form QH2 (reduced).
2- Complex II (Succinate dehydrogenase complex)
• It is a flavoprotein composed of succinate
dehydrogenase bound to FAD.
• It catalyzes the transfer of 2 hydrogens from
succinate to ubiquinone to form fumarate and QH2.
• Other flavoproteins can donate their hydrogen to
ubiquinone.
3- Complex III (Cytochrome bc1 complex)
• It is a hemoprotein composed mainly of cytochrome b
and c1.
• It catalyzes the transfer of 2 electrons from QH2 to
cytochrome c. At this site 2 hydrogen ions (2H+) are
released and two electrons flow along the successive
cytochromes till they meet oxygen to form water.
4- Complex IV (Cytochrome c Oxidase)
• It is a hemoprotein composed of cytochrome a-a3 and
2 atoms of copper.
• It catalyzes the transfer of electrons from cytochrome c
to oxygen. At this complex oxygen, electrons and the
hydrogen ions (previously released from QH2) are
combined to form H2O.
 P : O Ratio

• Definition:
• It describes the relationship between ATP
generation and oxygen consumption
• It is the ratio of the number of ATP synthesized (or
phosphate utilized) per atom of oxygen consumed
(to form water) when electrons flow in the
respiratory chain from reduced coenzymes to O2.
• In case of oxidation of NADH the P:O ratio is 2.5
• While in case of oxidation of a substrate by
flavoprotein (FAD) dehydrogenase the P:O ratio is
1.5
 Regulation of Electron Transport Chain
Activity
Chemiosmotic Theory of ATP Synthesis by
Outer membrane ETC
Inter-membrane
space
4H+ 4H +
2H + 4H+

Q Cyt c
Inner ATP
[O]2-
membran III IV synthas
I
e II e
NADH,H 2H+ 4H+
+
NAD+ FADH2 FAD H2O
Matrix 4H+ 4H +
2H ADP + Pi
+
ATP
• ETC to work requires a source of hydrogen
(reduced NAD+ and FAD) and oxygen.
• The electrochemical potential difference across
the inner mitochondrial membrane once
established as a result of proton translocation,
inhibits further transport of reducing equivalents
(electrons) through the respiratory chain unless
discharged by back-translocation of protons
across the membrane through the ATP synthase.

• This in turn depends on the availability of ADP

and Pi (ADP is the activator of the ATP synthase)
During work,
 ATP utilization is increased with increased production of
ADP (↓ ATP/ADP ratio).
 ADP activates ATP synthase.
 Decrease proton back–pressure on the complexes of the
ETC.
 Activation of the proton pumps.
 Increase the rate of oxidation of NADH and ATP synthesis.
During rest,
 ATP utilization is decreased (↑ATP/ADP ratio),
 Inhibition of ATP synthase.
 Accumulation of protons in the inter-membrane space.
 Increase the back-pressure on the proton pumps Leads to
their inhibition.
 Decrease the rate of oxidation of NADH and ATP synthesis.
 Uncouplers
• Dissociate or uncouple oxidation in respiratory
chain from phosphorylation. So, the oxidation takes
place without ATP synthesis.
• Happens If protons could pass across the inner
mitochondrial membrane without passing through
Complex V ATP synthase.
• So energy liberated is lost in the form of heat
 Uncouplers
Example of Physiologic uncouplers:
•Thermogenesis: It is a mean of generating heat to maintain body
temperature in some animals (including humans) on exposure to cold.
This occurs mainly in brown adipose tissue

Brown adipose tissue: It is very rich in mitochondria, and it is

specialized for this process of thermogenesis. The inner mitochondrial
membrane of this tissue contains a large amount of thermogenin, which
called the uncoupling protein (UCP). Thermogenin forms a channel
for the passage of protons from the inter-membrane space to the
matrix (bypasses the ATP synthase) and causes the release of the
energy of the gradient as heat without ATP synthesis.
 Uncouplers
•Exposure to cold results in sympathetic stimulation. This
leads to release of norepinephrine (noradrenaline) which binds
to the β-adrenergic receptor in the cell membrane of brown
adipose tissue and results in a cascade of events ending in:
activation of hormone sensitive lipase→ hydrolysis of
triacylglycerol→ release of FFAs→ activation of thermogenin
 Uncouplers
Examples of pathologic Uncouplers:
1- 2, 4-dinitrophenol:
Obsolete drug was used previously in the treatment
of obesity.
2- Thyroxin: only in cases of Thyrotoxicosis
(Hyperthyroidism)
They act by increasing the inner mitochondrial
membrane permeability to the protons.
Brown Adipose tissue & Thermogenesis
 CITRIC ACID CYCLE
Tricarboxylic Acid Cycle (TCA cycle)

Krebs' Cycle
 Krebs' Cycle

 Definition:
It is series of reactions that are responsible for
the complete oxidation of acetyl-CoA.
 Krebs' Cycle
 Definition:
It is series of reactions that are responsible for
the complete oxidation of the acetyl moiety of
acetyl-CoA.
It is the final common pathway for the oxidation
of carbohydrates, lipids and proteins
During the oxidation of acetyl-CoA,
☑CO2 is produced
☑coenzymes (NAD+ and FAD) are reduced and
subsequently reoxidized in the respiratory chain with
the formation of ATP & water.
 Krebs' Cycle
 Site:
All enzymes of the TCA cycle are found in the
Mitochondrial Matrix except
Succinate Dehydrogenase
 CH3– CO ~ S-CoA
CoA- SH
Active acetate CH2– COOH
O=C- COOH HO–C– COOH
CH2-COOH CH2– COOH
Oxaloacetate Citrate synthase Citrate
H2O
Aconitase
NADH,H+ Fe2+
Malate dehydrogenase G-SH H2O
NAD + TCA Cycle
CH2– COOH
HO–CH–COOH
C– COOH
CH2– COOH
CH– COOH
L-Malate Cis-aconitate

Fumarase Aconitase H2O

H2O Fe2+
HC – COOH G-SH
CH2– COOH
HOOC –CH
Fumarate ETC 9 ATP HC– COOH
HOCH– COOH
Isocitrate
FADH2
Succinate NAD+
dehydrogenase Isocitrate
FAD dehydrogenase
CH2– COOH NADH,H+
CH2– COOH
Succinate CH2– COOH
HC– COOH
CoA-SH ATP O=C – COOH
Oxalosuccinate
Succinate thiokinase
Mg2+ Isocitrate
ADP + Pi dehydrogenase
CoA-SH Mn2+ CO2
CH2– COOH NADH,H+ -Ketoglutarate
NAD+ CH – COOH
CH2 dehydrogenase 2

O=C ~ S-CoA complex CH2

Succinyl-CoA O=C – COOH
TPP, Lipoate, & FAD
CO2 -Ketoglutarate
 Krebs' Cycle
 Steps
1) Formation of citrate
 Krebs' Cycle
 Steps
2) Formation of Isocitrate
 Krebs' Cycle
 Steps
3) Formation of α-ketoglutarate
 Krebs' Cycle
 Steps
4) Formation of succinyl-CoA
 Any α Keto acid can
undergo
Oxidative Decarboxylation

Removal of 2H Removal of CO2

To give next lower acyl CoA
(lesser in the number of carbons by 1 and carried by CoASH)
The process requires
 3 enzymes collectively called “----- dehydrogenase
complex”
 5 coenzymes they are “TPP, Lipoate, CoASH, FAD &
NAD”
 Krebs' Cycle
 Steps
5) Formation of succinate
 Krebs' Cycle
 Steps
6) Formation of fumarate
 Krebs' Cycle
 Steps
7) Formation of malate
 Krebs' Cycle
 Steps
8) Formation of oxaloacetate
 CH3– CO ~ S-CoA
CoA- SH
Active acetate CH2– COOH
O=C- COOH HO–C– COOH
CH2-COOH CH2– COOH
Oxaloacetate Citrate synthase Citrate
H2O
Aconitase
NADH,H+ Fe2+
Malate dehydrogenase G-SH H2O
NAD + TCA Cycle
CH2– COOH
HO–CH–COOH
C– COOH
CH2– COOH
CH– COOH
L-Malate Cis-aconitate

Fumarase Aconitase H2O

H2O Fe2+
HC – COOH G-SH
CH2– COOH
HOOC –CH
Fumarate ETC 9 ATP HC– COOH
HOCH– COOH
Isocitrate
FADH2
Succinate NAD+
dehydrogenase Isocitrate
FAD dehydrogenase
CH2– COOH NADH,H+
CH2– COOH
Succinate CH2– COOH
HC– COOH
CoA-SH ATP O=C – COOH
Oxalosuccinate
Succinate thiokinase
Mg2+ Isocitrate
ADP + Pi dehydrogenase
CoA-SH Mn2+ CO2
CH2– COOH NADH,H+ -Ketoglutarate
NAD+ CH – COOH
CH2 dehydrogenase 2

O=C ~ S-CoA complex CH2

Succinyl-CoA O=C – COOH
TPP, Lipoate, & FAD
CO2 -Ketoglutarate
 Krebs' Cycle
 Importance of Citric Acid Cycle
1. It is a common pathway for oxidation of carbohydrates,
fats and amino acids

Carbohydrates Proteins Lipids

Stage One

Monosaccharides Amino Acids Glycerol + Fatty Acids

Stage Two

Acetyl-CoA

Stage Three
Oxidized H2O + ATP +
Coenzymes Free Energy
Krebs’Cycle
Reduced
Coenzymes [O] + ADP + Pi
CO2
 Krebs' Cycle
 Importance of Citric Acid Cycle
2. Energy production:
• The oxidation of one mole of the acetyl group of acetyl-CoA
through Krebsʹ cycle yields 10 moles of ATP.
 Krebs' Cycle
 Regulation of Citric Acid Cycle
• In the Krebs' cycle there are three irreversible steps.
• These three reactions of the cycle are catalyzed by
 citrate synthase,
 isocitrate dehydrogenase and
 α-ketoglutarate dehydrogenase
• which are the rate controlling enzymes of the cycle
(key enzymes)
 Krebs' Cycle
 Regulation of Citric Acid Cycle
I. Substrate activation and product feedback inhibition
 Oxaloacetate and acetyl-CoA
 Oxaloacetate
 succinyl-CoA
 CH3– CO ~ S-CoA
CoA- SH
Active acetate CH2– COOH
O=C- COOH HO–C– COOH
CH2-COOH CH2– COOH
Oxaloacetate Citrate synthase Citrate
H2O
Aconitase
NADH,H+ Fe2+
Malate dehydrogenase G-SH H2O
NAD + TCA Cycle
CH2– COOH
HO–CH–COOH
C– COOH
CH2– COOH
CH– COOH
L-Malate Cis-aconitate

Fumarase Aconitase H2O

H2O Fe2+
HC – COOH G-SH
CH2– COOH
HOOC –CH
Fumarate ETC 9 ATP HC– COOH
HOCH– COOH
Isocitrate
FADH2
Succinate NAD+
dehydrogenase Isocitrate
FAD dehydrogenase
CH2– COOH NADH,H+
CH2– COOH
Succinate CH2– COOH
HC– COOH
CoA-SH ATP O=C – COOH
Oxalosuccinate
Succinate thiokinase
Mg2+ Isocitrate
ADP + Pi dehydrogenase
CoA-SH Mn2+ CO2
CH2– COOH NADH,H+ -Ketoglutarate
NAD+ CH – COOH
CH2 dehydrogenase 2

O=C ~ S-CoA complex CH2

Succinyl-CoA O=C – COOH
TPP, Lipoate, & FAD
CO2 -Ketoglutarate
 Krebs' Cycle
 Regulation of Citric Acid Cycle
II- NADH/NAD+ ratio
III- ATP/ ADP ratio (Energy Requirements)
IV- During muscular exercise
During Work
↓ATP → ↑ ADP → Activation of ATP synthase → Activation of
oxidation by ETC → ↓NADH → ↑NAD → Activation of TCA cycle
key enzymes

During Rest
↑ATP → ↓ADP → Inhibition of ATP synthase → Inhibition of
oxidation by ETC → ↑NADH → ↓NAD → Inhibition of TCA cycle
key enzymes