Krebs Cycle

INTRODUCTION
The Krebs cycle is the central metabolic hub of the
cell.
This cycle is also citric acid cycle because the cycle
begins with the formation of citric acid which
containing 3 COOH groups.
It is the final common pathway for the oxidation of fuel
molecule such as amino acids, fatty acids, and
carbohydrates.
In, eukaryotes, the reactions of the citric acid cycle
take place inside mitochondria, in contrast with those
of glycolysis, which take place in the cytosol.

 Described by Hans Adolf Krebs in 1937.

A feature of cell chemistry shared by all types of life.
A complex series of reactions beginning and ending with the
compound oxaloacetate.
The cycle produces carbon dioxide and the energy-rich
compound ATP.
Eight successive reaction steps.
The six carbon citrate is formed from two carbon acetyl-CoA
and four carbon oxaloacetate.
Oxidation of citrate yields CO2 and regenerated
oxaloaxetate.
The energy released is captured in the reduced coenzymes
NADH and FADH2.

 The four-carbon molecule, oxaloacetate that initiates the

first step in the citric acid cycle is regenerated at the end
of one passage through the cycle.
The oxaloacetate acts catalytically: it participates in the
oxidation of the acetyl group but is itself regenerated.
Thus, one molecule of oxaloactetate is capable of
participating in the oxidation of many acetyl molecules.

Role of oxaloacetate

CONVERSION OF
PYUVATE TO ACEYL CoA

Pyruvate enters the mitochondrion from the cytoplasm.

One carbon atom is removed via decarboxylation and hydrogen is
removed using NAD+.
Pyruvate is converted into acetyl-CoA by the pyruvate
dehydrogenase complex.
Coenzyme A becomes attached to the remaining carbon atoms,
creating acetyl-CoA, which then enters the Krebs cycle.

Step 1: Formation of Citrate

Oxaloacetate first condenses with acetyl CoA to

form citryl CoA, which is then hydrolyzed to
citrate and CoA.
This reaction is catalyzed by the enzyme, which
is known as citrate synthase.

Step 2: Formation of Isocitrate

Citrate is isomerized into isocitrate to enable the
six-carbon unit to undergo oxiadative
decarboxylation
The isomerization of citrate is accomplished by a
dehydration step followed by a hydration step.
The result in an interchange of a hydrogen atom
and a hydroxyl group.
The enzyme catalyzing both steps is called
aconitase because cis-aconitase is an intermediate.

Aconitase is an iron-sulfur protein, or nonheme iron

protein. It contains four iron atoms that are not
incorporated as part of a heme group.
The poison Fluoroacetate is toxic, because fluroacetylCoA condenses with oxaloacetate to form fluorocitrate,
which inhibits Aconitase, causing citrate to accumulate.
The mode of inhibition is suicidal inhibition.

Step 3: Formation of -ketoglutarate

Isocitarte undergoes dehydragenation catalyzed by
isocitrate dehydrogenase to form, initially
oxalosuccinate, which remains enzyme-bound and
undergoes decarboxylation to -ketoglutarate.
The decarboxylation requires Mg2+ or Mn2+ ions.
There are three isoenzymes of isocitrate
dehydrogenase.
One, which uses NAD+, is found only in mitochondria.
The other two use NADP+ and are found in
mitochondria and the cytosol.

Respiratory chain-linked oxidation of isocitrate

proceeds almost completely through the NAD+dependent enzyme.
In this experiment, one CO2 is eliminated.
Hence, the -ketoglutarate has only 5 carbon
atoms.

Step 4: Formation of
Succinyl CoA

The conversion of isocitrate into -ketoglutarate

is followed by a second oxidative
decarboxylation reaction, the formation of
Succinyl CoA from -ketoglutarate.

The enzyme, which is -ketoglutarate

dehydrgenase is responsible in this reaction.
Two hydrogen atoms are released and they are
transferred to NAD+. The NAD+ is converted into
NADH.
The -ketoglutarate dehydrogenase complex
requires the same cofactors as the pyruvate
dehydrogenase complex thiamine diphosphate,
lipoate, NAD+ , FAD, and CoA- and result in the
formation of succinyl-CoA.

Step 5: Formation of Succinate

The succinyl CoA is decarboxylated to succinate
which yields one ATP per succinyl-CoA.
The cleavage of the thioester bond of succinyl
CoA is coupled to the phophorykation of a purine
nucleoside diphosphate, usually GDP.
This step is catalysed by succinyl-CoA synthetase.
CoA is liberated and displaced by a phosphate
group, which is transferred to GDP, forming GTP,
a molecule with function similar to ATP

This is the only example in the citric acid

cycle of substrate level phosphorylation.

Step 6: Formation of Fumarate

Succinate is oxidised to fumarate by the removal of 2

hydrogen atoms, which is bound to the inner surface of
the inner mitochondrial membrane.
The reaction is catalysed by succinate dehydrogenase.
The hydrogen atoms are accepted by FAD and it forms
FADH2.

Step 7: Formation of Malate

Fumerate is then converted to malate by hydration with

the use of fumerase.
Addition of a water molecule rearranges bonds in the
substrates.

Step 8: Regeneration of
oxaloacetate

It is the final step in Krebs cycle. Oxaloacetate is

regenerated from malate by a process of dehydrogenation.
This reaction is catalysed by malate dehydrogenase in the
presence of NAD. The 2 hydrogen atoms removed are
accepted by NAD and it forms NADH.
The oxaloacetate formed in the above reaction condenses
with the acetyl CoA to form citric acid again and thus the
cycle is repeated.

Summary

 two molecules of pyruvate

2 ATP molecules
10 NADH molecules
two FADH2 molecules as well as
six CO2 molecules in the form of
waste gas

The end result is