Bioenergetics

is the branch of biochemistry that focuses on how cells

transform energy, often by producing, storing or consuming
adenosine triphosphate (ATP). Bioenergetic processes, such as
cellular respiration or photosynthesis, are essential to most
aspects of cellular metabolism, therefore to life itself.
 Lesson Outline
ENERGY REQUIREMENTS FOR METABOLISM

GLYCOLYSIS

THE TRICARBOXYLIC ACID ( TCA) cycle

OXIDATIVE PHOSPHORYLATION IN THE

RESPIRATORY CHAIN
 I. ENERGY REQUIREMENTS FOR METABOLISM
Metabolism- encompasses the chemical processes
used to synthesize complex molecules from basic
precursor molecules (anabolism) and to break
down complex molecules into less complex
components (catabolism).

Anabolism-building large molecules

from smaller ones.
( consumes energy)

Catabolism- is the breakdown of large molecules

into smaller molecules.
(Generates energy)
 concept of free energy
• Every living cell and organism must perform work to stay alive, to grow and to reproduce. The energy
processes in living organisms are defined by the basic laws of thermodynamics.
The energy actually available to do work (utilizable) is known as free energy.
• Changes in the free energy (AG) are valuable in predicting the feasibility of chemical reactions.
• The reactions can occur spontaneously if they are accompanied by decrease in free energy.
• During a chemical reaction, heat may be released or absorbed.

free energy of a chemical reaction depends on the heat energy and entropy of the reactants and products. Free
energy also depends on the concentration of reactants and products.
 concept of free energy

In biochemistry, the change in Gibbs free energy under standard conditions (when all components are at a
concentration of 1 molar, 25° C, 1 atm pressure, and a pH of 7) is indicated by AG°
• Cells are isothermal systems, meaning they function at a constant temperature & pressure.
• Photosynthetic cells acquire free energy from absorbed solar radiation.
• Heterotrophic cells acquire free energy from nutrient molecules.
• Cells transform this free energy into ATP & other energy-rich compounds to provide energy for biological
work.
Types of reactions in free energy
 Types of reactions in free energy

Equilibrium- the rate of the forward reaction equals the rate of the reverse reaction.

AG=0
 SOURCES OF ENERGY

• The hydrolysis of adenosine triphosphate (ATP) provides the main source of free energy in
biochemical reactions.
The triphosphate group of ATP contains two high-energy phosphoanhydride bonds that, when
hydrolyzed, each release approximately 7.3 kcal/mol of free energy; AG°' for ATP hydrolysis to
adenosine diphosphate (ADP) = -7.3 kcal/mol
 ANOTHER SOURCES OF ENERGY
• Nucleoside triphosphates (NTPs) such as guanosine-, uridine-, and cytidine triphospahte ( GTP, UTP, AND
CTP, respectively) also yield free energy through the hydrolysis of their phosphoanydride bonds.
• a source of energy for cellular reactions and are involved in signalling pathways.
• The nucleoside triphosphates have the sugar deoxyribose;

• Another high energy molecule , acethyl CoA transfer acethyl group in a way that is analogous to the
transfer of phosphoryl groups by ATP
• Acetyl-CoA is a metabolite derived from glucose, fatty acid, and amino acid catabolism.

CELLS REPLINISH ATP THROUGH THREE PATHWAYS

• GLYCOLYSIS-Glycolysis is the process in which glucose is broken down to produce energy. It produces two molecules of pyruvate, ATP, NADH
and water. The process takes place in the cytoplasm of a cell and does not require oxygen. It occurs in both aerobic and anaerobic organisms.
The pyruvate- can be used in the citric acid cycle or serve as a precursor for other reactions.
is a three-carbon acid that is naturally formed during glycolysis, the process in which the body breaks down sugar (glucose).

ANAEROBIC- NO
OXYGEN IS USED
 CELLS REPLINISH ATP THROUGH THREE PATHWAYS

2. The Krebs cycle starts with the products of glycolysis, which are two three-carbon
molecules known as pyruvate.

The Krebs cycle or TCA cycle (tricarboxylic acid cycle) or Citric acid cycle-cells first
convert pyruvate to acetyl coenzyme A (acetyl Co) and then oxidize it completely to
CO2; the oxidation of two pyruvate molecules provides enough energy to form two
molecules of the high-energy compound GTP, which can be converted to ATP

3.Oxidative phosphorylation-involving the electron transport chain takes place in the

inner mitochondrial membrane. The electron transport chain is a collection of proteins
bound to the inner mitochondrial membrane and organic molecules, which electrons
pass through in a series of redox reactions, and release energy

AEROBIC- OXYGEN IS
USED
 RELEASE OF ENERGY BY OXIDATION FOODSTUFFS

The human body obtains the energy needed for all its metabolic processes through the oxidation of
carbohydrates, fats, and proteins in food.

what is oxidation? When oxygen combines with an element or compound, an oxidation reaction occurs.
Oxidation can also be defined as the process of the removal of hydrogen from the reactant species. Oxidation is
the process of losing electrons by a molecule, atom, or ion.
what is foodstuffs? Carbohydrates, proteins, and lipids are the three most frequent types of molecules found in food. These
molecules are also known as 'macronutrients,' and they are nutritionally important to us. The three groupings of molecules
have quite different properties that will affect how your meal turns out.

This process involving oxidation of food to release energy to form of ATP is called Cellular respiration.
Respiration involves breakdown of glucose. It produces carbon, water and releases energy in the form of ATP.
It is a biochemical process and takes place inside the cell. The released energy is utilized for various metabolic
activities.
 REGULATION OF METABOLISM

•Metabolism regulation involves intricate processes that maintain the balance between energy intake and expenditure
in living organisms. Hormones, such as insulin and glucagon, play key roles in regulating metabolism. Insulin
promotes glucose uptake by cells, lowering blood sugar levels, while glucagon raises blood sugar levels by
stimulating the liver to convert stored glycogen into glucose. Additionally, metabolic rate is influenced by factors like
physical activity, diet, and genetics. Disruptions in this regulation can lead to metabolic disorders like diabetes and
obesity.
• Prokaryotes regulate metabolism by synthesizing enzymes based on immediate cellular needs, adjusting to the
availability of specific substrates.
• Eukaryotes regulate metabolism through enzyme control, separate pathways for anabolism and catabolism, and
physical segregation of metabolic reactions. Additionally, the energy charge of a cell, determined by ATP, AMP,
and ADP levels, governs the synthesis or utilization of ATP to maintain cellular energy balance.
 II.GLYCOLYSIS

GLYCOLYSIS- GLYCO + LYSIS

GLUCOSE- 2 PYRUVATE
2 C-C-C

C-C-C-C-C-C

ATP & NAHD

GLYCOLYSIS
2 AT P- C OMPL E T E OX IDAT IO N O F T WO
MOL E C UL E S O F PYR U VAT E TO C O

2 PYR UVAT E - C onv ersion o f o n e m o lecu le o f.

GLUCOSE glu co se to two mo lecu les o f p y ru v ate

Oxidative phosphorylation-Produces 32 or 34 molecules

 GLUCOSE

PYRUVIC ACID

OXYGEN PRESENT? NO OXYGEN

LACTIC ACID ETHANOL

KREBS CYCLE (ANIMALS) (YEAST)

OXIDATIVE PHOSPHORYLATION
 • in ALL GLYCOlYSIS, ALL THE INTERMEDIATE
MOLECULES BETWEEN GLUCOS ( THE STARTING
COMPOUND ) AND PYRUVATE ( THE FINAL PRODUCT )
ARE PHOSPHORYLATED; THE PHOSPHATE IS ADDED
ASAN ESTER OR AN ANHYDRIDE; ANHYDRIDE BONDS
ARE USUALLY HIGH-ENERGY BONDS.
 WHAT IS GLYCOLYSIS ?

Glycolysis- is a nearly universal pathway in prokaryotes and eukaryotes, is the series of enzyme catalyzed
reactions by which one molecule of glucose is converted to two molecules of pyruvate, two ATP molecules are
produced.

• the complete oxidation of glucose through glycolysis, the TCA cycle, and oxidation phosphorylation produces
energy that is stored in the form of ATP.
• in glycolysis, all the intermediate molecules between glucose ( the starting compound) and pyruvate or lactate
( the final product) are phosphorylated; the phosphate group is added as an ester or ananhydride; anhydride bonds
are usually high energy bonds.

The glycolytic pathway- the multistep process of glycolysis converts one molecule of glucose to two molecule of
pyruvate , the conversion of pyruvate to lactate generally occurs only in anaerobic conditions.
THE GLYCOLYTIC PATHWAY

• Priming Stage- the first series of reactions in glycolysis, convert

glucose to fructose-1, 6-biphosphate.
• Splitting stage- the second series of reactions in glycolysis, splits
the six-carbon compound fructose 1 , 6-biphosphate into two
three-carbon compounds ( eliminately two molecules of
glyceraldehyde 3-phosphate.
• The oxideration- phosphorylation stage- converts the two
molecules glyceraldehyde 3-P two molecules of pyruvate ( or
lactate, under anaerobic conditions), and produces for molecule
of ATP.
• in feast and certain bacteria, pyruvate is converted to ethanol
and CO2 by anaerobic fermentation.
III. THE TRICARBOXYLIC ACID ( TCA) CYCLE
 GLYOCXYLATE CYCLE

•The glyoxylate cycle is a variant of the tricarboxylic acid (TCA) cycle found in certain microorganisms and plants.
Unlike the conventional TCA cycle, the glyoxylate cycle enables these organisms to bypass two decarboxylation steps,
conserving carbon during the metabolism of fatty acids or other carbon compounds. This cycle allows the conversion of
acetyl-CoA into oxaloacetate, which can then be used for gluconeogenesis or for replenishing the TCA cycle
intermediates. The glyoxylate cycle plays a significant role in organisms that rely on acetate or fatty acids as a carbon
source, allowing them to efficiently utilize these carbon compounds for energy production and biosynthesis.

The glyoxylate cycle, found in plants and bacteria, diverges from the conventional TCA cycle by allowing two acetyl units
to enter simultaneously. In this cycle, isocitrate generated from one acetyl unit is cleaved into succinate and glyoxylate,
bypassing the TCA cycle's decarboxylation steps. Glyoxylate then reacts with the second acetyl unit to form malate, which
is further oxidized to oxaloacetate. This oxaloacetate serves as a precursor for gluconeogenesis, enabling the synthesis of
glucose from non-carbohydrate sources, a crucial process for these organisms' energy and biosynthesis needs.
 IV. OXIDATIVE PHOSPHORYLATION IN THE RESPIRATORY CHAIN

General Information

1.Oxidative phosphorylation is a process whereby ATP is produced as electrons are shuttled through the energy-generating
components of the respiratory chain (also called the electron transport chain) to molecular oxygen (O). forming water.

2. During oxidative phosphorylation, the oxidation of each NADH produces the molecules of ATP, the oxidation of each
FADH, produces two molecules of ATP.

3. Oxidative phosphorylation is the major source of ATP for aerobic organisms

4. In eukaryotes, oxidative phosphorylation occurs in the mitochondria membrane; in prokaryotes, it occurs in the
cytoplasmic membrane.
 IV. OXIDATIVE PHOSPHORYLATION IN THE RESPIRATORY CHAIN

Energetics of oxidative phosphorylation

The Mechanism of Oxidative Phosphorylation

Most of the usable energy obtained from the breakdown of carbohydrates or fats is derived by
oxidative phosphorylation, which takes place within mitochondria. For example, the breakdown of
glucose by glycolysis and the citric acid cycle yields a total of four molecules of ATP, ten molecules
of NADH, and two molecules of FADH2 (see Chapter 2). Electrons from NADH and FADH2 are
then transferred to molecular oxygen, coupled to the formation of an additional 32 to 34 ATP
molecules by oxidative phosphorylation. Electron transport and oxidative phosphorylation are critical
activities of protein complexes in the inner mitochondrial membrane, which ultimately serve as the
major source of cellular energy.
 IV. OXIDATIVE PHOSPHORYLATION IN THE RESPIRATORY CHAIN

Respiratory chain components

The repiratory chain is a collection of enzymes that induce the electrochemical gradient of protons (H+),
necessary for the ATP synthesis. Fatty acid, amino acid and monosaccharide metabolism yields acetyl- CoA, a
molecule that is oxidized in the Citric acid cycle producing reduced coenzymes (NADH+H+, FADH2) and GTP.
These coenzymes are subsequently collected and transported by the repsiratory chain, directing them to their final
reaction with O2 to form H2O. The respiratory chain consists of enzymes that are found only in the inner
membrane of mitochondria, thus only mitochondrion-containing cells are able to produce energy through
oxidative phosphorylation (aerobic respiration). Mitochodrionless cells, such as erythrocytes, produce energy only
through glycolysis, an anaerobic process of glucose oxidation into pyruvate.
Oxidative phosphorylation
It is the process through which the liberated free energy provided by the electrochemical gradient of protons (H+), is
trapped as high energy phosphate. The electrochemical built-up of hydrogen ions, by the respiratory chain is used to drive
protons through an enzyme called ATP synthase. The kinetic energy of protons is used by the synthase to store energy as
high energy phosphate in ATP by binding ADP and Pi.

Components of the respiratory chain

Electrons flow through the repsiratory chain, passing through three large protein enzyme complexes.
NADH-Q oxidoreductase/Complex I: the electrons are transported from the NADH+H+ to coenzyme Q Q-Cytochorme c
oxidoreductase/Complex III: coenzyme Q passes the electrons to the cytochrome c
Cytochrome c oxidase/Complex IV:completes the chain, passing the electrons to O2 and causing it to be reduced to H2O
Succinate-Q reductase/Complex II:the electrons are transferred from FADH2 to coenzyme Q
The flow of electrons through the respiratory chain generates ATP through the process of oxidative phosphorylation. Enzyme complexes
I, III and IV act as proton pumps which cause the accumulation of protons in the intermembrane space of mitochondria constituting it
more positive than the intramembrane space (matrix) which becomes progressively more negative building an electrochemical gradient
with protons hving the tendency to return back into the matrix which is more negative. The only way to do that is passing through the
ATP synthase which exploits their energy to form ATP.
Inhibitors

Inhibitors of the respiratory chain:

Barbiturates: they inhibit complex I/NADH-Q oxidoreductase, preventing electron transfer from NADH+H+ to coenzyme
Q

CO2, H2S and CN-: they inhibit complex IV/cytochrome c oxidase preventing reduction of O2 to H2O
Antimycin A: it inhibits complex III/Q-cytochrome c oxidoreductase preventing transfer of electrons from CoQ to cyt c.

Inhibitors of the oxidative phosphorylation: atractyloside which inhibits ATP synthase by blocking the flow of protons
through the F0 subunit. Uncouplers of oxidative phosphorylation: 2,4 dinitrophenol which uncouples proton pumping
from ATP synthesis because it carries protons across the inner mitochondrial membrane (no ATP synthase inhibiton, rather
elimination of the proton electrochemical gradient built-up)
 GENERATION OF ATP BY PROTON-MOTIVE FORCE

Proton Motive Force

• The transfer of H+ through a proton pump generates an electrochemical
gradient of protons.

1. The flow of electrons from NADH to is a thermodynamically favorable reaction

2. Recall that a mitochondrion has to primary membranes, an outer membrane and

an inner membrane that invaginates into a central space called matrix; the space
between the inner membrane and the outer membrane is called the intermembrane
space

3.During the synthesis of ATP, protons flow from the intermembrane space back
into the matrix through special ion channels; these channels are a structural feature
of the enzyme complex that synthesis ATP, called ATPase or ATP synthase.

4. ATPase, located in the inner mitochondrial membrane, is composed of different

protein subunits
 GENERATION OF ATP BY PROTON-MOTIVE FORCE

5. The chemiosmotic hypothesis describes how the proton gradient between the mitochondrial matrix and the
intermembrane space results in production of ATP

RESPIRATORY CHAIN INHIBITORS AND

UNCOUPLERS

1.The three pumping sites of the respiratory chain can be blocked by various inhibitors

2. Continued exposure to respiratory chain inhibitors leads to death of the organism from insufficient energy
production

3. Oligomycin (an antibiotic) inhibits mitochondrial ATPase directly

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