BIO462

BIOCHEMISTRY

THE CITRIC ACID

CYCLE

Lesson Learning Outcomes

Upon completion of this chapter, students should be

able to:
1. explain the steps of the citric acid cycle
2. differentiate between citric acid cycle and glyoxylate

cycle
3. relate citric acid cycle as an energy source

INTRODUCTION
Also known as Kreb cycle or tricarboxylic acid cycle

(TCA cycle)
Three processes play central roles in aerobic
metabolism
the citric acid cycle (this chapter)
electron transport
oxidative phosphorylation

Metabolism consists of
catabolism: the oxidative breakdown of nutrients
anabolism: the reductive synthesis of biomolecules

The citric acid cycle is amphibolic; that is, it plays a

role in both catabolism and anabolism

Catabolism
Fats

Anabolism

Polysaccharides

beakdown
of larger
molecules Fatty acids Monosacto smaller and glycerol charides
ones
oxidation and the
release of energy

Proteins
Amino
Acids

Small
Anabolism
molecules of proteins

Excretion

Anabolism

Catabolism

Excretion

Products of anabolism,
including proteins and
nucleic acids
energy and
reducing
agents
Some nutrients and
products of catabolism

The role of ATP as an energy source

The Citric Acid Cycle

Where citric acid cycle

Takes place in the matrix of mitochondria except
happens?
for one in which the intermediate electron
acceptor is FAD (inner mitochondrail membrane)

In the citric acid cycle and the pyruvate

dehydrogenase reactions, one molecule of pyruvate

is oxidized to 3 molecules of CO2 as a result of
oxidative phosphorylation.
The oxidations are accompanied by reductions.
4 NAD+ are reduced to NADH
1 FAD is reduced to FADH2
1 GDP is phosphorylated to GTP

Pyruvate to Acetyl-CoA
Step 1: pyruvate loses CO2 and HETPP is formed
pyruvate
O
dehydrogenase
CH3 CCOO + TPP
Pyruvate

OH
CO2 + CH3 CH-TPP
Hydroxyethyl-TPP

Step 2: requires lipoic acid

the active form of lipoic acid is bound to the enzyme

by an amide bond to the amino group of a lysine

COOH

reduction
oxidation

S S
Lipoic acid

COOH
HS

SH
Dihydrolipoic acid

Pyruvate to Acetyl-CoA
Step 3: the acetyl group is transferred to the

sulfhydryl group of coenzyme A

O
C-NH-Enz

SH
CoA-SH + CH3 C-S
Coenzyme A
Dihydrolipoamide
dihydrolipoyl transacylase
O
C-NH-Enz

O
CoA-S-CCH3
Acetyl-CoA

+ HS

SH
Dihydrolipoamide

Pyruvate to Acetyl-CoA
Step 4: oxidation of dihydrolipoamide
O
C-NH-Enz
HS

SH

Dihydrolipoamide
NAD+
NADH
O
C-NH-Enz

S S

Lipoamide

The Citric Acid Cycle

Step 1: condensation of acetyl-CoA with

oxaloacetate;
O
CH3 C-SCoA
Acetyl-CoA citrate
synthase
+
O C-COO
CH2 -COOOxaloacetate

CH2 -COO-

HO C-COO- + CoA-SH
CH2 -COO- Coenzyme A
Citrate

The Citric Acid Cycle

Step 2: dehydration and rehydration

gives isocitrate; catalyzed by aconitase

CH2 -COOHO C-COO-

CH2 -COOC-COO-

CH2 -COO
Citrate

CH-COOAconitate

CH2 -COOH C-COOHO CH-COOIsocitrate

only one of the 4 stereoisomers of

isocitrate is formed in the cycle

The Citric Acid Cycle

Step 3: oxidation of isocitrate followed

by decarboxylation

CH2 -COO- NAD+ NADH

H C-COO-

HO CH-COO
Isocitrate

isocitrate
dehydrogenase
CH2 -COOH C-COOO C-COOOxalosuccinate

CO2

CH2 -COOH C-H

O C-COO-Ketoglutarate

isocitrate dehydrogenase is an allosteric

enzyme; it is inhibited by ATP and NADH,

activated by ADP and NAD+

The Citric Acid Cycle

Step 4: oxidative decarboxylation of -

ketoglutarate to succinyl-CoA
CoA-SH
CH2 -COOCH2
O C-COO-Ketoglutarate

NAD+

NADH

-ketoglutarate
dehydrogenase
complex

CH2 -COOCH2

+ CO2

O C SCoA
Succinyl-CoA

like pyruvate dehydrogenase, this enzyme is

a multienzyme complex and requires

coenzyme A, thiamine pyrophosphate, lipoic
acid, FAD, and NAD+

The Citric Acid Cycle

Step 5: formation of succinate
CH2-COOCH2

+ GDP + Pi

O C SCoA

Succinyl-CoA

succinyl-CoA
synthetaseCH2-COO-

CH2-COO

+GTP + CoA-SH

Succinate

this is the first energy-yielding step of the

cycle
the overall reaction is slightly exergonic

The Citric Acid Cycle

Step 6: oxidation of succinate to

fumarate
CH2 -COO-

FAD

CH2 -COO-

succinate
dehydrogenase

Succinate

FADH2
-

OOC

C
C

COOH

Fumarate

Step 7: hydration
of fumarate
H
COO
C
HO
CH-COO
H2 O
C
CH
-COO
fumarase
2
OOC
H
Fumarate
L-Malate

The Citric Acid Cycle

Step 8: oxidation of malate
-

HO CH-COO

CH2 -COOL-Malate

NAD NADH O C-COOmalate

dehydrogenase

CH2 -COOOxaloacetate

From Pyruvate to CO2

Pyruvate dehydrogenase complex
Pyruvate+CoA-SH+NAD+
+
Acetyl-CoA
+ NADH+CO2+ H
Citric acid cycle
+
Acetyl-CoA
+3NAD +FAD+GDP+Pi +2H2O
2CO2 +CoA-SH+3NADH+ 3H++FADH2 +GTP
+
Pyruvate+4NAD+FAD+ GDP+ Pi + 2H2O

3CO2 +4NADH+FADH2 +GTP+ 4H

Control of the CA Cycle

Three control points within the cycle
citrate synthase: inhibited by ATP, NADH, and

succinyl CoA; also product inhibition by citrate

isocitrate dehydrogenase: activated by ADP
and NAD+, inhibited by ATP and NADH
-ketoglutarate dehydrogenase complex:
inhibited by ATP, NADH, and succinyl CoA;
activated by ADP and NAD+

One control point outside the cycle

pyruvate dehydrogenase: inhibited by ATP and

NADH; also product inhibition by acetyl-CoA

Control of the CA Cycle

The Glyoxylate Cycle

Plants and some bacteria, but not animals, use a

modification of the citric acid cycle to produce

four-carbon dicarboxylic acids and eventually
glucose

the glyoxylate cycle bypasses the two oxidative

decarboxylations of the citric acid cycle

instead, it routes isocitrate via glyoxylate to malate
key enzymes in this cycle are isocitrate lyase and
malate synthase

Glyoxylate Cycle

The Glyoxylate Cycle

The glyoxylate cycle takes place
in plants: in glyoxysomes, specialized organelles

devoted to this cycle

in yeast and algae: in the cytoplasm

Helps plants grow in the dark

seeds are rich in lipids, which contain fatty acids
during germination, plants use the acetyl-CoA

produced in fatty acid oxidation to produce

oxaloacetate and other intermediates for carbohydrate
synthesis
once plants begin photosynthesis and can fix CO2,
glyoxysomes disappear

CA Cycle in Catabolism
The catabolism of proteins, carbohydrates, and

fatty acids all feed into the citric acid cycle at one
or more points
Proteins
Amino Acids

Carbohydrates

Fatty Acids

Pyruvate
Acetyl-CoA
-Ketoglutarate
Succinyl-CoA
Fumarate
Malate
Oxaloacetate

intermediates
of the citric
acid cycle

CA Cycle in Anabolism
The citric acid cycle is the source of starting

materials for the biosynthesis of other compounds

examples:

O
-

OOCCH2 CH2 CCOO

transamination

NH3 +
-

OOCCH2 CH2 CHCOOGlutamate

-Ketoglutarate
O
transamination
OOCCH2 CCOO
Oxaloacetate

NH3 +
-

OOCCH2 CHCOOAspartate