Lec#33-Lec#36

Introduction to TCA cycle

Citric Acid cycle
Introduction
The citric acid cycle (Krebs cycle or
tricarboxylic acid-TCA cycle) is the most
important metabolic pathway for the energy
supply to the body. About 65-70% of the ATP is
synthesized in Krebs cycle. Citric acid cycle
essentially involves the oxidation of acetyl CoA
to CO2 and H2O.
TCA- the central metabolic
pathway
• The citric acid cycle is the final common
oxidative pathway for carbohydrates, fats and
amino acids.
This cycle not only supplies energy but also
provides many intermediates required for the
synthesis of amino acids, glucose, heme etc.
Krebs cycle is the most important central pathway
connecting almost all the individual metabolic
pathways (either directly or indirectly).
Brief History
• The citric acid cycle
was proposed by
Hans Adolf Krebs in
1937, based on the
studies of oxygen
consumption in
pigeon breast muscle.
• The cycle is named in
his honour (Nobel
Prize for Physiology
and Medicine in 1953.)
Location of TCA CYCLE

• The enzymes of TCA cycle are located in

mitochondrial matrix, in close proximity to
the electron transport chain.
This enables the synthesis of ATP by
oxidative phosphorylation without any
hindrance.
TCA - is an open cycle
• Krebs cycle is a cyclic process. However, it
should not be viewed as a closed circle, since
many compounds enter the cycle and leave. TCA
cycle is comparable to a heavy traffic circle in a
national highway with many connecting roads.
Each intermediate of the cycle connecting
another pathway is a road!
ROLE
• The cycle oxidizes pyruvate (formed
during the glycolytic breakdown of glucose)
to CO2 and H2O, with the concomitant
production of energy.
• Acetyl CoA from fatty acid breakdown and
amino acid degradation products are also
oxidized.
• In addition, the cycle has a role in
producing precursors for biosynthetic
pathways.
Location

• The citric acid cycle occurs within the

mitochondria of eukaryotes and the
cytosol of prokaryotes.
The citric acid cycle has eight stages:
1. Production of citrate from oxaloacetate and acetyl CoA (catalyzed by
citrate synthase).
2. Isomerization of citrate to isocitrate (catalyzed by aconitase).

3. Oxidation of isocitrate to  ketoglutarate (catalyzed by isocitrate

dehydrogenase; the reaction requires NAD+).

4. Oxidation of  ketoglutarate to succinyl CoA (catalyzed by the

 ketogluterate dehydrogenase complex; the reaction requires NAD+).

5. Conversion of succinyl CoA to succinate [catalyzed by succinyl CoA

synthetase; the reaction requires inorganic phosphate and GDP (or
ADP).

6. Oxidation of succinate to fumarate (catalyzed by succinate

dehydrogenase; the reaction involves FAD).

7. Hydration of fumarate to malate (catalyzed by fumarase).

8. Oxidation of malate to oxaloacetate (catalyzed by malate

dehydrogenase
 aconitase
Citrate
Synthase

malate isocitrate
dehydrogenase. dehydrogenase

fumarase

succinate
dehydrog
enase
succinyl CoA  ketoglutarate
synthetase dehydrogenase
The citric acid cycle
The cycle carries out the oxidation of acetyl groups from acetyl CoA to CO2 with the production of four
pairs of electrons, stored initially in the reduced electron carriers NADH and FADH2
The cycle has eight stages:
1. Citrate (6C) is formed from the irreversible condensation of acetyl CoA (2C) and oxaloacetate (4C) –
catalyzed by citrate synthase.
2. Citrate is converted to isocitrate (6C) by an isomerization catalyzed by aconitase.
This is actually a two-step reaction during which cis-aconitate is formed as an intermediate. It is the
cis-aconitate which gives the enzyme its name.
3. Isocitrate is oxidized to  ketoglutarate (5C) and CO2 by isocitrate dehydrogenase. This
mitochondrial enzyme requires NAD+, which is reduced to NADH.
4.  Ketoglutarate is oxidized to succinyl CoA (4C) and CO2 by the  ketoglutarate dehydrogenase
complex. Like pyruvate dehydrogenase, this is a complex of three enzymes and uses NAD+ as a
cofactor.
5. Succinyl CoA is converted to succinate (4C) by succinyl CoA synthetase. The reaction uses the
energy released by cleavage of the succinyl–CoA bond to synthesize either GTP (mainly in
animals) or ATP (exclusively in plants) from Pi and, respectively, GDP or ADP.
6. Succinate is oxidized to fumarate (4C) by succinate dehydrogenase. FAD is tightly bound to the
enzyme and is reduced to produce FADH2.
7. Fumarate is converted to malate (4C) by fumarase; this is a hydration reaction requiring the addition
of a water molecule.
8. Malate is oxidized to oxaloacetate (4C) by malate dehydrogenase. NAD+ is again required by the
enzyme as a cofactor to accept the free pair of electrons and produce NADH.
Step 3 – Oxidation of NADH and FADH2 produced by the citric acid cycle
The NADH and FADH2 produced by the citric acid cycle are reoxidized and the energy released is
used to synthesize ATP by oxidative phosphorylation
Energy Yield
• Each of the three NADH molecules produced per turn of
the cycle yields 3 ATPs and the single FADH2 yields 2
ATPs by oxidative phosphorylation (although some
measurements indicate that the quantities are 2.5 and
1.5 respectively.
• One GTP (or ATP) is synthesized directly during the
conversion of succinyl CoA to succinate.
Thus the oxidation of a single molecule of glucose via the
citric acid cycle produces 12 ATP molecules.
Role of TCA Cycle
 Regulation of TCA
The cellular demands of ATP are crucial in
controlling the rate of citric acid cycle. The
regulation is brought about either by enzymes or
the levels of ADP.
Three enzymes-namely :
citrate synthase, isocitrate dehydrogenase and
 -ketoglutarate dehydrogenase-regulate citric acid cycle.
1. Citrate synthase is inhibited by ATP, NADH, acetyl CoA and
succinyl CoA.
2. lsocitrate dehydrogenase is activated by ADP, and inhibited
by ATP and NADH.
3.  Ketoglutarate dehydrogenase is inhibited by succinyl CoA
and NADH.
4. Availability of ADP is very important for
the citric acid cycle to proceed. This is due to
the fact that unless sufficient levels of ADP are
available, oxidation (coupled with phosphorylation
of ADP to ATP) of NADH and FADH2
through electron transport chain stops.
The accumulation of NADH and FADH2 will lead to
inhibition of the enzymes (as stated above) and
also limits the supply of NAD+ and FAD which
are essential for TCA cycle to proceed.