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Metabolism:
Citric Acid Cycle

461

Cellular Respiration
Molecular processes by which cells
consume O2 and produce CO2
Three Major Stages
▪ Stage 1 - organic fuel molecules—glucose,
fatty acids, and some amino acids—are
oxidized to yield two-carbon fragments in
the form of the acetyl group of acetyl-
coenzyme A (acetyl-CoA).

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Cellular Respiration
Molecular processes by which cells
consume O2 and produce CO2
Three Major Stages
▪ Stage 2 - the acetyl groups are fed into
the citric acid cycle, which enzymatically
oxidizes them to CO2; the energy
released is conserved in the reduced
electron carriers NADH and FADH2

463

Cellular Respiration
Molecular processes by which cells
consume O2 and produce CO2
Three Major Stages
▪ Stage 3 - the reduced coenzymes are
themselves oxidized, giving up protons
(H+) and electrons. The electrons are
transferred to O2—the final electron
acceptor—via a chain of electron-carrying
molecules known as the respiratory chain.

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The Citric Acid Cycle

Tricarboxylic acid cycle; Krebs cycle
▪ series of reactions that oxidizes the acetyl group of acetyl-CoA to
two molecules of CO2 in a manner that conserves the liberated
free energy for utilization in ATP generation.
▪ It accounts for the major portion of carbohydrate, fatty acid, and
amino acid oxidation and generates numerous biosynthetic
precursors
▪ Amphibolic – operates both catabolically and anabolically

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The Citric Acid Cycle

Tricarboxylic acid cycle; Krebs cycle
▪ Amphibolic – operates both
catabolically and anabolically

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The Citric Acid Cycle

Production of Acetyl-CoA
▪ Before entering the citric acid
cycle, the carbon skeletons of
sugars and fatty acids are
degraded to the acetyl group
of acetyl-CoA
▪ Pyruvate is oxidized to acetyl-
CoA and CO2 by the pyruvate
dehydrogenase (PDH)
complex

467

The Citric Acid Cycle

Production of Acetyl-CoA
The Pyruvate Dehydrogenase Complex Requires Five Coenzymes
▪ thiamine pyrophosphate (TPP),
▪ flavin adenine dinucleotide (FAD),
▪ coenzyme A (CoA)
▪ nicotinamide adenine dinucleotide (NAD)
▪ lipoate

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The Citric Acid Cycle

Production of Acetyl-CoA
The Pyruvate Dehydrogenase Complex Requires Five Coenzymes
▪ coenzyme A (CoA)

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The Citric Acid Cycle

Production of Acetyl-CoA
The Pyruvate Dehydrogenase
Complex Requires Five Coenzymes
▪ lipoate

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The Citric Acid Cycle

Production of Acetyl-CoA
The Pyruvate Dehydrogenase Complex Consists of Three Distinct
Enzymes

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The Citric Acid Cycle

Production of Acetyl-CoA
The Pyruvate Dehydrogenase Complex Consists of Three Distinct
Enzymes

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The Citric Acid Cycle

Production of Acetyl-CoA
The Pyruvate Dehydrogenase Complex
Consists of Three Distinct Enzymes
▪ E1 catalyzes first the decarboxylation
of pyruvate, producing hydroxyethyl-
TPP, and then the oxidation of the
hydroxyethyl group to an acetyl
group.
▪ The electrons from this oxidation
reduce the disulfide of lipoate bound
to E2, and the acetyl group is
transferred into thioester linkage with
one OSH group of reduced lipoate.

473

The Citric Acid Cycle

Production of Acetyl-CoA
The Pyruvate Dehydrogenase
Complex Consists of Three Distinct
Enzymes
▪ E2 catalyzes the transfer of the
acetyl group to coenzyme A,
forming acetyl-CoA.

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The Citric Acid Cycle

Production of Acetyl-CoA
The Pyruvate Dehydrogenase
Complex Consists of Three Distinct
Enzymes
▪ E3 catalyzes the regeneration of
the disulfide (oxidized) form of
lipoate; electrons pass first to FAD,
then to NAD+

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The Citric Acid Cycle

Production of Acetyl-CoA Occurs in Five Reactions

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Reactions of the Citric Acid Cycle

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Reactions of the Citric Acid Cycle

(1) Formation of Citrate
▪ the condensation of acetyl-CoA
with oxaloacetate to form citrate
▪ Enzyme: citrate synthase
▪ The CoA liberated in this reaction
is recycled to participate in the
oxidative decarboxylation of
another molecule of pyruvate by
the PDH complex

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Reactions of the Citric Acid Cycle

(2) Formation of Isocitrate via cis-
Aconitate
▪ Enzyme: aconitase
▪ reversible transformation of
citrate to isocitrate, through the
intermediary formation of the
tricarboxylic acid cis-aconitate.
▪ Aconitase can promote the
reversible addition of H2O to the
double bond of enzyme-bound
cis-aconitate

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Reactions of the Citric Acid Cycle

(3) Oxidation of Isocitrate to α-Ketoglutarate and CO2
▪ Enzyme: isocitrate dehydrogenase
▪ Oxidative decarboxylation of isocitrate to form α-ketoglutarate

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Reactions of the Citric Acid Cycle

(4) Oxidation of α-Ketoglutarate to
Succinyl-CoA and CO2
▪ Enzyme: α-ketoglutarate
dehydrogenase
▪ oxidative decarboxylation in which
α-ketoglutarate is converted to
succinyl-CoA and CO2
▪ NAD+ serves as electron acceptor
and CoA as the carrier of the
succinyl group

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Reactions of the Citric Acid Cycle

(5) Conversion of Succinyl-CoA to
Succinate
▪ Enzyme: succinyl-CoA synthetase
▪ energy released in the breakage
of the thioester bond is used to
drive the synthesis of a
phosphoanhydride bond in either
GTP or ATP

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Reactions of the Citric Acid Cycle

(6) Oxidation of Succinate to
Fumarate
▪ Enzyme: succinate
dehydrogenase
▪ oxidation of succinate’s central
single bond to a trans double
bond, yielding fumarate with the
concomitant reduction of the
redox coenzyme FAD to FADH2

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Reactions of the Citric Acid Cycle

(7) Hydration of Fumarate to Malate
▪ Enzyme: fumarase
▪ The reversible hydration of
fumarate to L-malate

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Reactions of the Citric Acid Cycle

(8) Oxidation of Malate to
Oxaloacetate
▪ Enzyme: L-malate dehydrogenase
▪ Reformation of oxaloacetate by
oxidizing malate’s secondary
alcohol group to the corresponding
ketone with concomitant reduction
of a third NAD+ to NADH.

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The Citric Acid Cycle

Functions of the TCA Cycle
▪ Glucose biosynthesis which occurs
in the cytosol, utilizes oxaloacetate
as its starting material.
▪ Oxaloacetate is not transported
across the mitochondrial
membrane, but malate is.
▪ Malate that has been transported
across the mitochondrial
membrane is converted to
oxaloacetate in the cytosol for
gluconeogenesis.

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The Citric Acid Cycle

Functions of the TCA Cycle
▪ Lipid and cholesterol biosynthesis
▪ a cytosolic process that requires
acetyl-CoA.
▪ Acetyl-CoA is generated in the
mitochondrion and is not transported
across the inner mitochondrial
membrane.
▪ Cytosolic acetyl-CoA is therefore
generated by the breakdown of
citrate, which can cross the inner
mitochondrial membrane, in a
reaction catalyzed by ATP-citrate
lyase

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The Citric Acid Cycle

Functions of the TCA Cycle
▪ Amino acid biosynthesis utilizes
citric acid cycle intermediates in
two ways.
▪ α-Ketoglutarate is converted to
glutamate in a reductive amination
reaction involving either NAD+ or
NADP+ catalyzed by glutamate
dehydrogenase
▪ α-Ketoglutarate and oxaloacetate
are also used to synthesize
glutamate and aspartate in
transamination reactions

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Regulation of the Citric Acid Cycle

Rate-limiting Enzymes
▪ Citrate Synthase
▪ Isocitrate Dehydrogenase
▪ α-Ketoglutarate Dehydrogenase

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Regulation of the Citric Acid Cycle

Rate-limiting Enzymes
Citrate Synthase
▪ Citrate is a competitive inhibitor of
oxaloacetate for citrate synthase
(product inhibition);
▪ Succinyl-CoA also competes with
acetyl-CoA

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Regulation of the Citric Acid Cycle

Rate-limiting Enzymes
Isocitrate Dehydrogenase
▪ inhibited by NADH and ATP

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Regulation of the Citric Acid Cycle

Rate-limiting Enzymes
α-Ketoglutarate Dehydrogenase
▪ Inhibited by its products, NADH
and succinyl-CoA

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