Lipid Metabolism

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• Lipid metabolism involves breakdown and
synthesis of lipid: cholesterol, fatty acids,
triacylglycerol, phospholipids etc.

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 Fatty acid oxidation (β-oxidation)

• Fatty oxidation occurs in the mitochondria but

synthesis occurs in the cytosol.
• It is an aerobic process requiring the presence of oxygen
• Increased fatty acid oxidation is a characteristic of
starvation and of diabetes mellitus, leading to ketone
body formation by the liver (ketosis).
• Because gluconeogenesis is dependent on fatty acid
oxidation , any impairment of fatty acid oxidation leads
to hypoglycaemia

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Steps in Fatty Acid Degradation and Synthesis

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Oxidation of Fatty Acids occurs in the Mitochondria

• Fatty-acid oxidation begins with activation of the

molecule
• Fatty acids are activated before being catabolized

• First converted to an active intermediate before they can

be catabolized .
• Only step in the complete degradation of fatty acids that
requires energy from ATP.
• ATP is converted to AMP and Ppi.
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• In the presence of ATP and CoA, the enzyme acyl-CoA
synthetase (thiokinase) catalyzes the conversion of a fatty
acid to an ‘’active fatty acid’’ or acyl-CoA which uses
high energy phosphate with the formation of AMP and
Ppi.

• Long chain fatty acids do not cross the inner membrane as

acyl-CoA but as carnitine derivatives

• Short and medium chain fatty acids enter the mitochondria

passively
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 β-Oxidation of Fatty Acids involves successive
cleavage with release of Acetyl-CoA

• In β-oxidation, two carbons at a time are cleaved from

acyl-CoA molecules, starting at the carboxyl end

• The cyclic reaction sequence generates FADH2 and

NADH

• The chain is broken between α (2) and β (3)-carbon

atoms, hence the name β-oxidation.

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Reaction Sequence for the Degradation of Fatty Acids

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10
• Long-chain acyl-CoA is cycled between reactions 2 to 5,
acetyl-CoA being split off, each cycle by thiolase. When
acyl radical is 4 carbon atoms in length, two acetyl-CoA
molecules are formed in the reaction 5.

Oxidation of fatty acids produces a large quantity of

ATP

• Eg Palmitate- a 16-carbon fatty acid

FADH2 and NADH will result in 4 high energy
phosphates in the respiratory chain for each of the first 7
acetyl-CoA molecules formed by β-
oxidation (7 X 4 =28)
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• A total of 8 molecules of acetyl-CoA is formed and each
will give 12 molecules of ATP on oxidation in the CAC.
Total (8 X 10 = 80)

• One is subtracted from the initial activation of fatty acid

yielding a net gain of 107 molecules of ATP per mole of
palmitate

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 Biosynthesis of Fatty Acids

• The main pathway for de novo synthesis of fatty

acids (lipogenesis) occurs in the cytosol

• The system is present in many tissues including liver,

kidney, brain, lung, mammary gland, and adipose
tissue

• Acetyl-CoA is the precursor and free palmitate is

the end product

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• Production of malonyl-CoA is the initial and
controlling step in fatty acid synthesis

• Bicarbonate is a source of CO2 in the carboxylation

of acetyl-CoA to malonyl-CoA in the presence of
ATP and acetyl-CoA carboxylase

• Acetyl-CoA carboxylase - requires biotin

- is a multienzyme protein
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Acetyl-CoA is the principal building block of fatty acids

Fatty acid synthesis

Pyruvate Glucose
Fatty acids

Mitochondrial
membrane
Pyruvate
dehydrogenase
Acetyl-CoA
Acetyl-CoA
Oxaloacetate

ATP-citrate lyase NADH

Malate
Oxaloacetate dehydrogenase
Citrate
Citrate NAD+
Malate
CAC NADP+
Malic enzyme
Pyruvate NADPH*
carboxylase Pyruvate Pyruvate
CO2 CO2
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Fatty Acid Synthesis.

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• The fatty acid synthase complex catalyzes formation
of fatty acids
▪ Is a dimer
▪ Each polypeptide contains seven enzyme activity

• Acetyl-CoA used as a primer forms carbon atoms 15

and 16

• Addition of all the other C2 units is via malonyl-CoA

• Propionyl-CoA acts as a primer for synthesis of long-

chain fatty acids having odd number of carbon atoms
-ruminant fat and milk
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 Acetyl –CoA
Carboxylase
Malonyl-CoA
BIOTIN, Mn
Malonyl-CoA
transacylase
Acetyl
transacylase

ENZYME Acetyl-ACP-malonyl-ACP
COMPLEX
3 Ketoacyl -
synthase
3-Ketoacyl-ACP

NADPH + H+ 3 Ketoacyl
reductase

NADP+
D-3-Hydroxyacyl-ACP

From PPP,
isocitrate Hydratase
dehydrogenase,
2,3-Enoyl-ACP
malic enzyme
Enol reductase
NADPH + H+

NADP+
Acyl-ACP (e.g. buturyl-ACP)
Thioesterase
After cycling through
steps 2-5 seven times 18
Palmitate
The main source of NADPH for lipogenesis is the
pentose phosphate pathway

• The oxidative reactions of the Pentose Phosphate

Pathway (PPP) are the chief source of hydrogen
required for reductive synthesis of fatty acids

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• Tissues specializing in active lipogenesis ie liver,
adipose tissue, and lactating mammary gland also
possess an active PPP – both occur in the cytosol and
so there are no barriers against the transfer of NADPH

• Other sources of NADPH

- reaction that converts malate to pyruvate (catalysed
by NADP malate dehydrogenase ‘‘ Malic enzyme’’;
extramitochondrial isocitrate dehydrogenase )

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Elongation of fatty acids occurs in the endoplasmic
reticulum

• This pathway elongates saturated and unsaturated fatty

acyl-CoAs (from C10 upwards) by two carbon units

• Malonyl-CoA as acetyl donor and NADPH as reductant

• Catalyzed by fatty acid elongase system of enzymes

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 Ketone bodies are formed from acetyl-
CoA when fat breakdown dominates
- Acetyl-CoA formed from fatty oxidation
enters the CAC only if fat and carbohydrate
degradation are approximately balanced.
- Entry of acetyl-CoA into the CAC depends
on the availability of oxaloacetate for the
formation of citrate; but concentration of
oxaloacetate is lowered if carbohydrate is
unavailable or improperly utilized;
oxaloacetate is normally formed from
pyruvate, the product of glycolysis 22
• In fasting or diabetics oxaloacetate is consumed to
form glucose by the gluconeogenic pathway and
hence unavailable for condensation with acetyl-CoA

• Under these conditions acetyl-CoA is diverted to the

formation of acetoacetate, D-3-hydroxybutyrate and
acetone - the ketone bodies

• Abnormally high levels of ketone bodies are present

in the blood of untreated diabetics

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Formation of Ketone Bodies

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Ketone bodies serve as fuel for extrahepatic
tissues
• In extrahepatic tissues acetoacetate is
activated to acetoacetyl-CoA by transferase
(NB liver lacks this enzyme)

• CoA is transferred from succinyl-CoA to

form acetoacetyl-CoA

• The acetoacetyl-CoA is split to acetyl-CoA

by thiolase and oxidized in the CAC 25
• While acetoacetate and D-3-hydroxybutyrate
are readily oxidized in the extrahepatic
tissues, acetone is difficult to oxidize in vivo
and is to a large extent volatilized in the
lungs (i.e. exhaled)
• Heart muscle and renal cortex tend to use
acetoacetate for energy in preference to
glucose.
• During starvation and in diabetes the brain
adapts to utilization of acetoacetate for
energy supply
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Transport of ketone bodies from liver and pathways of utilization and
oxidation in extrahepatic tissues

MUSCLE
FFA

Acetyl-CoA
LIVER
Acyl-CoA Thiolase
Acetoacetyl-CoA

Acetyl-CoA
Succinate OAA
CoA
HMG-CoA transferase Citrate
succinyl-CoA
CO2
Acetoacetate
Acetoacetate
NADH + H+ NADH + H+
NAD+ NAD+
3-hydroxybutyrate
3-hydroxybutyrate

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 The nutritional state regulates lipogenesis

• Excess of carbohydrate is stored as fat in many animals in

anticipation of periods of caloric deficiency.
• Lipogenesis converts surplus glucose and intermediates
such as pyruvate, lactate and acetyl-CoA to fats
• Its depressed under low restricted caloric intake, or when
there is deficiency of insulin – the later conditions are
associated with increased concentrations of free fatty acids
in the plasma

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• The nutritional state is the main factor
regulating the rate of lipogenesis
• Lipogenesis is increased when sucrose is
fed instead of glucose because fructose
bypasses the phosphofructokinase
control point in glycolysis and floods
the lipogenic pathway

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Short and long term mechanisms regulate lipogenesis

• Long-chain fatty acid synthesis is controlled in the

short term by allosteric and covalent modification of
enzymes and in the long term by changes in gene
expression governing rates of enzyme synthesis

• Acetyl carboxylase is the most important enzyme in

the regulation of lipogenesis
Activated by - citrate
-Insulin (via insulin-stimulated
protein kinase)
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Inhibited by:
- phosphorylation of the enzyme (
stimulated by the hormones Glucagon
and epinephrine)
- long-chain acyl-CoA molecules (negative
feedback inhibition)
Insulin also regulates lipolysis by other mechanisms
- Increases transport of glucose into the cell (adipose
tissue), increasing the availability of pyruvate for fatty
acid synthesis and glycerol 3-phosphate for
esterification

- Activates pyruvate dehydrogenase in adipose tissue but

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not in liver