BIOLOGICAL OXIDATION

Biological oxidation
The oxidation of food stuffs occurs in three stages

i. Primary metabolism – Digestion

ii. Secondary metabolism - Metabolism of bio molecules

iii. Tertiary metabolism – Electron transport chain

Reducing equivalents/Electron carriers

• NADH + H
• FADH2
 Biological oxidation

The transfer of electrons from the reduced coenzymes (NADH

+ H & FADH2) through the respiratory chain to oxygen is
known as Biological oxidation.

All the enzymes involved in the process of biological oxidation

belong to the major class of oxidoreductases.
sub class : Dehydrogenases, oxidases, oxygenases &
hydroperoxidases.

Coenzymes : NAD , FAD & FMN

 BIOENERGETICS
Study of energy changes in biochemical reactions

I. Exergonic Reaction - Energy releasing

II. Endergonic Reaction - Energy Consuming

Transfer of free energy from an exergonic to an

endergonic reaction via high energy intermediate
compound (~ E)

ATP acts as a “energy currency” of the cell,

transferring free energy……..
 High energy compounds

• Certain compounds are encountered in the biological system

which, on hydrolysis, yields energy.
• High energy compounds : Compounds when on hydrolysed
liberate more energy than that of ATP (– 7.3kcal/mol ).
• Low energy compounds : Compounds when on hydrolysed
liberate less energy than that of ATP (– 7.3kcal/mol ).
• Most of the high-energy compounds contain phosphate group
(exception acetyl CoA) hence they are called high-energy
phosphate compounds.
• The high-energy compounds possess acid anhydride bonds
STANDARD FREE ENERGY OF HYDROLYSIS OF COMPOUNDS

ENERGY OF
HYDROLYSIS
OF COMPOUNDS
 High energy compounds
Significance :

I. ATP required for phosphorylation, synthetic

reactions.
II. GTP used for protein synthesis
III. UDP-Glucose required for glycogen and lactose
synthesis.
IV. Succinyl CoA , used for substrate phosphorylation.
V. Creatine phosphate energy for muscle contraction.
 Cellular respiration

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 NADH shuttle
• Inner mitochondrial membrane is Impermeable to
NADH
• NADH produced in the cytosol is entering into
mitochondria through membrane shuttle system.
• There are two shuttle to transport cytosolic NADH
to mitochondria

I. Glycerophosphate shuttle
II. Malate shuttle
Glycerophosphate shuttle
Malate shuttle
 Biological oxidation

Oxidation : It is defined as loss of electrons.

Reduction : It is defined as gain of electrons.

Redox couple :
substance exists both in the reduced state and in the
oxidised state, the pair is called a redox couple.

Redox potential:[EO] Electron transfer potential of this

redox couple is known as redox potential.
Redox potentials
 Biological oxidation
• Negative redox potential
Substances having lower affinity for electrons

• Positive redox potential

Substances having higher affinity for electrons
 REDOX POTENTIAL

The five complexes of electron transport chain

are arranged in a order of Increasing REDOX
POTENTIAL
ELECTRON TRANSPORT CHAIN
 Components of ETC

Complex I - NADH: coenzyme Q reductase

Complex II - Succinate : coenzyme Q reductase
Complex III - Cytochrome reductase
Complex IV - Cytochrome oxidase
Complex V - ATP synthase
 Components of ETC
• Iron Sulphur Proteins :
These are non- heme iron containing proteins
and are also called Rieske iron sulphur
protein.

• Cytochrome b :
It has two heme residues in one polypeptide.
 Mobile electron carrier of ETC

I. Coenzyme Q or ubiquinone is a lipid soluble

compound functions as a mobile electron carrier
within the mitochondrial inner membrane.
Reduced form- ubiquinol (UQH2)
I. oxidised form - ubiquinone (UQ)

II. Cytochrome C : It is water soluble compound and

contains one heme group.
 Electron transport chain

•Succinate dehydrogenase
•Fatty acyl-CoA dehydrogenase
•Glycerol phosphate dehydrogenase

•Glyceraldehyde 3 phosphate
dehydrogenase
•Isocitrate dehydrogenase
•Malate dehydrogenase
•Glutamate dehydrogenase
•Beta hydroxy acylcoA
dehydrogenase

SITES OF OXIDATIVE PHOSPHORYLATION

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 OXIDATIVE PHOSPHORYLATION
Oxidative phosphorylation is the series of energy
transformations occurring in mitochondria involving
transformation of electron-motive force into a
proton-motive force and, finally, the proton-motive
force is used for Phosphorylation of ADP to ATP
 1961 | 1978
Peter Mitchell
• Proposed chemiosmotic hypothesis

proton motive force

1920-1992

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 Mechanism of oxidative phosphorylation

CHEMIOSMOTIC THEORY

I. The flow of electrons from NADH or FADH2 to O2 through electron

transport chain located in the mitochondrial inner membrane
leads to the pumping of protons out of the mitochondrial matrix.
II. The resulting uneven distribution of protons generates a pH
gradient and a transmembrane electrical potential that creates
a proton-motive force.
III. ATP is synthesized when protons flow back to the mitochondrial
matrix through an enzyme complex. Thus, the oxidation of fuels
and the phosphorylation of ADP are coupled by a proton gradient
across the inner mitochondrial membrane.
IV. This proton motive force drives the synthesis of ATP by ATP
synthase complex.

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 Mechanism of oxidative phosphorylation

For complete rotation of F1 head through the 3 states , 10 protons are translocated.

I. Complex I Pumps 4 protons (H+)

II. Complex III pumps 4 protons (H+)
III. Complex IV Pumps 2 protons (H+)
For every 4 protons (H+) 1 ATP is synthesized
NADH = 2.5 ATP
FADH2 = 1.5 ATP
 ATP synthase – Complex - V

I. It Is embedded In the Inner mitochondrial

membrane and looks like a ball on a stick

II. ATP synthase composed of a proton -

conducting unit and a catalytic unit.

III. Proton conducting unit is called FO and the

catalytic unit which protrudes into the
mitochondrial matrix is called F1

IV. The FI unit consists of five subunits

(α3β3γδϵ)

V. The proton gradient driven rotation of the C

ring drives the rotation of gamma subunit
The world’s smallest molecular motor in 120 – degree , which in turn promotes the
synthesis of ATP through the binding change
mechanism.
 ATP synthase

Synthesis of ATP by Binding change mechanism

L- loose binding ; T – Tight binding and O- open

The 3 catalytic sites on the enzyme ATP synthase bind ADP and phosphate in
sequence and then undergo a conformational change so as to make a tightly
bound ATP. The sites then change conformation again to release the ATP.
 P: O Ratio

It is defined as the number of ADP molecules

Phosphorylated to ATP for every atom of
oxygen consumed.

P:O ratio for NADH = 2.5

P:O ratio for FADH2 = 1.5
 Ratios regulating cellular respiration
• Electrons do not flow from fuel molecules to
oxygen unless ATP needs to be synthesized.
• The energy change , or more precisely ADP
concentration, normally determines the rate
of electron transport.
• The electrons flow down the respiratory
chain only when ADP levels are high

I. ATP/ADP Ratio
II. NADH/NAD Ratio
INHIBITORS OF ETC
 UNCOUPLERS

I. Uncouplers allow the electron transport to

proceed but prevent the coupling of electron
transfer to the phosphorylation of ADP.
II. It disturbs proton gradient across the
membrane.
III. It is biologically useful to newborn Infants.
 UNCOUPLERS

• Chemicals
• 2,4 dinirtophenol
• 2,4 dinitro cresol
• (CCP)Chlorocarbonyl cyanide phenyl hydrazone

• Physiological
• Thyroxine in high dozes
• Thermogenin are in brown adipose tissue
• UCP (Uncoupling protein) I,II,III
•I- adipose tissue
•II- Skeletal Muscle
•III-adipose tissue
 Ionophores

I. Ionophores are molecules that facilitate ion passage

in or out of cell membranes.
II. They are lipophilic molecules that complex specific
cations and facilitate their transport through biologic
membranes, eg, valinomycin (K+).
III. The classic uncouplers such as dinitrophenol are, in
fact, proton Ionophores.
Thank You