Fatty Acid Catabolism

Fatty Acids

 Fatty acids contains a long hydrocarbon chain and a terminal

carboxylate group.
 The hydrocarbon chain may be saturated or unsaturated.
Functions of fatty acids
Fatty acids have four major physiological roles:
1. They are the building blocks of phospholipids and glycolipids.
2. Many proteins are modified by the covalent attachment of
fatty acids, which target them to membrane locations
3. Fatty acids are fuel molecules, stored as triacylglycerol.
4. Fatty acid derivatives serve as hormones and intracellular
messengers eg steroids.
Catabolism pathways
Oxidation of fatty acids is a major
energy source in many organisms
• About 1/3 of our energy needs comes from dietary
triacylglycerols.

• 80% of the energy needs of mammalian heart and liver are met
by oxidation of fatty acids.

• Many hibernating animals, such as grizzy bears, rely almost

exclusively on fats as their source of energy (and water during
their long-term sleep)
Fats provide efficient fuel storage
• Advantages of fats over polysaccharides:
1. Fatty acids carry more energy per carbon because they are
more reduced.
2. Fatty acids carry less water along because they are nonpolar

• Glucose and glycogen are for short-term energy needs, quick

delivery. Fats are for long term (months) energy needs, good
storage and slow delivery.
 Energy from fatty acids
• When you eat fried food, you are consuming a lot of fat. The body is able
to gain a lot of energy from a single molecule of fat. For comparison, each
gram of fat (on average) gives the body 9 calories of energy while a gram
of protein or carbohydrates only gives 4 calories of energy.

• Or, to look at this in another way, 1 molecule of glucose will give the body
30-32 molecules of ATP (energy). While 1 molecule of palmitic acid (a 16
carbon fatty acid) will produce 160 molecules of ATP!

• The exact number of ATP produced from a molecule of fatty acid depends
on the type of fatty acid. Longer chain molecules will produce more ATP.
The process of creating energy from fatty acids is called fatty acid
oxidation, or beta oxidation.
Lipid metabolism is concerned mainly
with fatty acids and cholesterol

• The source of long-chain fatty acids is either dietary lipid or de

novo synthesis from acetyl-CoA derived from carbohydrate or
amino acids

• Fatty acids maybe oxidized to acetyl-CoA (Oxidation) or

esterified with glycerol, forming triacylglycerol (fat) as the body’s
main fuel reserve.
Acetyl-CoA formed by Beta-Oxidation
may undergo three fates:

• Acetyl-CoA arising from glycolysis oxidized to Co2 + H20 via

the citric acid cycle.

• It is the precursor for synthesis of cholesterol and other

steroids.

• In the liver, it is used to form ketone bodies (acetoacetate and

3-hydroxybutyrate) that are important fuels in prolonged fasting.
Much Amino Acid Metabolism involves
transamination

• Amino acids are required for protein synthesis.

• The essential amino acids must be supplied in the diet, since

they cannot be synthesized in the body.

• The nonessential amino acids are supplied in the diet, but can
also be formed from metabolic intermediates by transamination
using the amino nitrogen from other amino acids .
• After deamination, amino nitrogen is excreted as urea.

• The carbon skeletons that remain after transamination may:

1. Be oxidized to CO2 via the citric acid cycle

2. Be used to synthesize glucose (gluconeogenesis)
3. Form ketone bodies, which may be oxidized or be used for
synthesis of fatty acids.
Metabolic pathways studied at
different levels of organization

1. At the tissue and organ level the nature of the substrates

entering and metabolites leaving tissues and organs is
defined.

2. At the subcellular level each cell organelle (e.g mitochondria)

or compartment (e.g cytosol) has specific roles that form part
of a subcellular pattern of metabolic pathways.
AT THE TISSUE & ORGAN LEVEL, THE BLOOD
CIRCULATION INTEGRATES METABOLISM

• Amino acids resulting from the digestion of dietary protein are

absorbed via the hepatic portal vein.

• The liver has the role of regulating the blood concentration of

water-soluble metabolites.

• In the case of glucose, this is achieved by taking up glucose in

excess of immediate requirements and converting it to glycogen
(glycogenesis) or to fatty acids (lipogenesis).
• Between meals, the liver acts to maintain the blood glucose
concentration by breaking down glycogen (glycogenolysis) and,
together with the kidney, by converting noncarbohydrate
metabolites such as lactate, glycerol, and amino acids to glucose
(gluconeogenesis).

• The liver also synthesizes the major plasma proteins (eg,

albumin) and deaminates amino acids that are in excess of
requirements, forming urea, which is transported to the kidney
and excreted.
• Skeletal muscle utilizes glucose as a fuel, both aerobically,
forming CO2, and anaerobically, forming lactate.

• It stores glycogen as a fuel for use in muscle contraction and

synthesizes muscle protein from plasma amino acids.

• Muscle accounts for approximately 50% of body mass and

consequently represents a considerable store of protein that
can be drawn upon to supply amino acids for gluconeogenesis
in starvation.
• Lipids in the diet are mainly triacylglycerol, and are hydrolyzed
to mono-acylglycerols and fatty acids in the gut, then re-
esterified in the intestinal mucosa.

• Here they are packaged with protein and secreted into the
lymphatic system and thence into the bloodstream as
chylomicrons, the largest of the plasma lipoproteins.

• Chylomicrons also contain other lipid-soluble nutrients.

• Unlike glucose and amino acids, chylomicron triacylglycerol is not
taken up directly by the liver.

• It is first metabolized by tissues that have lipoprotein lipase, which

hydrolyzes the triacylglycerol, releasing fatty acids that are
incorporated into tissue lipids or oxidized as fuel.

• The chylomicron remnants are cleared by the liver.

• The other major source of long-chain fatty acids is synthesis

(lipogenesis) from carbohydrate, in adipose tissue and the liver.
• Adipose tissue triacylglycerol is the main fuel reserve of the
body.

• It is hydrolyzed (lipolysis) and glycerol and free fatty acids are

released into the circulation.

• Glycerol is a substrate for gluconeogenesis.

• The fatty acids are transported bound to serum albumin; they

are taken up by most tissues (but not brain or erythrocytes) and
either esterified to triacylglycerols for storage or oxidized as a
fuel.
Fatty acid breakdown

• Fatty acid breakdown (also called -oxidation) brings about the

oxidation of long-chain fatty acids with the production of energy
in the form of ATP.

• The fatty acids are converted into their acyl CoA derivatives and
then metabolized by the removal of two-carbon acetyl CoA units
from the end of the acyl chain.
Fatty acid breakdown involves a repeating sequence
of four reactions:
Activation
• FATTY ACIDS ARE ACTIVATED BEFORE BEING CATABOLIZED

• Fatty acid breakdown occurs in the cytosol of prokaryotes and in the

mitochondrial matrix of eukaryotes.
• Before entering the mitochondrial matrix, the fatty acid is activated by
forming a thioester link with CoA

• This reaction is catalyzed by acyl CoA synthase (also called fatty acid
thiokinase) which is present on the outer mitochondrial membrane, and
uses a molecule of ATP.
• The overall reaction is irreversible due to the subsequent hydrolysis of PPi
to two molecules of Pi.
Fatty Acid Transport into Mitochondria
• Fats are degraded into fatty acids and glycerol in the cytoplasm
of adipocytes.
• Fatty acids are transported to other tissues for fuel
• Beta-Oxidation of fatty acids occurs in the mitochondria.
• Small (<12 carbons) fatty acids diffuse freely across
mitochondrial membranes.
• Larger fatty acids (most free fatty acids) are transported via
acyl-carnitine/carnitine transporter (carnitine shuttle)
• This reaction, catalyzed by an enzyme on the outer face of the inner
mitochondrial membrane (carnitine acyltransferase I), removes the
CoA group and substitutes it with a carnitine molecule.

• The acylcarnitine is then transported across the inner mitochondrial

membrane by a carnitine/acylcarnitine translocase.

• This integral membrane transport protein transports acylcarnitine

molecules into the mitochondrial matrix and free carnitine molecules
out. Once inside the mitochondrial matrix the acyl group is
transferred back on to CoA, releasing free carnitine, by the enzyme
carnitine acyltransferase II which is located on the matrix side of
the inner mitochondrial membrane .
Beta-Oxidation pathway
• In animals the acetyl CoA produced from fatty acid degradation cannot be
converted into pyruvate or oxaloacetate.

• Although the two carbon atoms from acetyl CoA enter the citric acid cycle,
they are both oxidized to CO2 in the reactions catalyzed by isocitrate
dehydrogenase and -ketoglutarate dehydrogenase

• Thus, animals cannot convert fatty acids into glucose.

• In contrast, plants have two additional enzymes, isocitrate lyase and

malate synthase, that enable them to convert the carbon atoms of acetyl
CoA into oxaloacetate.

• This is accomplished via the glyoxylate pathway, a route involving

enzymes of both the mitochondrion and the glyoxysome, a specialized
membranous plant organelle.
Oxidation of Unsaturated fatty acids
• Require some additional processing before they can be unsaturated
degraded completely by -oxidation.

• Unsaturated fatty acyl CoAs with double fatty acids bonds at odd-
numbered carbon atoms (i.e. between, for example, C-9 and C- 10
as in palmitoleate; are acted on in the normal way by the
degradation mechanism until the acyl CoA dehydrogenase
encounters the cis-3-enoyl CoA formed at the end of the third round.

• The presence of the double bond between C-3 and C-4 prevents the
formation of another double bond between C-2 and C-3. To
overcome this problem an isomerase converts the cis-3 bond into a
trans-2 double bond, and the resulting trans-2-enoyl CoA can then
continue down the  -oxidation pathway
• Another enzyme, in addition to the isomerase, is required for
the oxidation of polyunsaturated fatty acids which have a
double bond at an even-numbered carbon atom.

• In this case the 2,4-dienoyl intermediate resulting from the

action of acyl CoA dehydrogenase is acted on by 2,4-dienoyl
CoA reductase to form cis-3-enoyl CoA .

• This is then converted by the isomerase into the trans form

which continues down the pathway.

• These reactions are important since over half the fatty acids of
plant and animal lipids are unsaturated (and often
polyunsaturated).
Ketone Bodies
When the level of acetyl CoA from -oxidation increases in excess of that required
for entry into the citric acid cycle, the acetyl CoA is converted into acetoacetate
and D-3-hydroxybutyrate by a process known as ketogenesis.

• D-3-hydroxybutyrate, acetoacetate and its nonenzymic breakdown product

acetone are referred to collectively as ketone bodies .

• Two molecules of acetyl CoA initially condense to form acetoacetyl CoA in a

reaction which is essentially the reverse of the thiolysis step in  -oxidation.

• The acetoacetyl CoA reacts with another molecule of acetyl CoA to form 3-
hydroxy-3-methylglutaryl CoA (HMG CoA) .
• This molecule is then cleaved to form acetoacetate and acetyl
CoA. (HMG CoA is also the starting point for cholesterol
biosynthesis.

• The acetoacetate is then either reduced to D-3-hydroxybutyrate

in the mitochondrial matrix or undergoes a slow, spontaneous
decarboxylation to acetone .

• In diabetes, acetoacetate is produced faster than it can be

metabolized. Hence untreated diabetics have high levels of
ketone bodies in their blood, and the smell of acetone can often
be detected on their breath.
• 3-HYDROXY-3-METHYLGLUTARYL-COA (HMG-COA) IS AN
INTERMEDIATE IN THE PATHWAY OF KETOGENESIS

• Enzymes responsible for ketone body formation are associated

mainly with the mitochondria.

• Two acetyl-CoA molecules formed in oxidation condense with one

another to form acetoacetyl-CoA by a reversal of the thiolase reaction.

• Acetoacetyl-CoA, (starting material for ketogenesis).

• Acetoacetyl-CoA arises directly from the terminal four carbons of a

fatty acid during β-oxidation.
Conversion of acetyl CoA to the ketone
bodies acetoacetate, acetone and D-3-
hydroxybutyrate
• Acetoacetate and D-3-hydroxybutyrate are produced mainly in
the liver and are not just degradation products of little
physiological value. They are used in preference to glucose as
an energy source by certain tissues such as the heart muscle
and kidney cortex.

• Although glucose is normally the major fuel for the brain, under
conditions of starvation or diabetes this organ can switch to
using predominantly acetoacetate
Fatty Acid synthesis
• Fatty acids are synthesized by the condensation of two-carbon
units.

• However, in terms of the enzymatic steps involved, the process

is not the reverse of -oxidation .

• Fatty acid synthesis involves a separate series of reactions to

build up long-chain hydrocarbons from acetyl CoA units
Key differences between fatty acid
synthesis and break down:
• Fatty acids are synthesized in the cytosol, but acetyl CoA is
produced from pyruvate in the mitochondria .
• Thus the acetyl CoA must be transferred from the mitochondria
into the cytosol to allow fatty acid synthesis to occur.
• However, the inner mitochondrial membrane is not readily
permeable to this molecule.
• This problem is overcome by the condensation of acetyl CoA
with oxaloacetate to form citrate .
• This is then transported into the cytosol where it is cleaved to
regenerate acetyl CoA and oxaloacetate by ATPcitrate lyase in
an energy-requiring process.
• The oxaloacetate, which also cannot cross the inner mitochondrial
membrane, is returned to the mitochondrial matrix through
conversion first to malate (catalyzed by malate dehydrogenase)
and then to pyruvate (catalyzed by NADP+-linked malate enzyme).

• This latter decarboxylation reaction generates NADPH which can be

used in fatty acid synthesis.

• The remaining NADPH required for fatty acid synthesis is provided

by the pentose phosphate pathway .

• Once back in the mitochondrial matrix, pyruvate is carboxylated to

form oxaloacetate by pyruvate carboxylase with the hydrolysis of a
further molecule of ATP .
The pathway of fatty acid biosynthesis
• The first committed step in fatty acid biosynthesis is the carboxylation of
acetyl CoA to form malonyl CoA using CO2 in the form of bicarbonate
HCO3

• This reaction is catalyzed by the enzyme acetyl CoA carboxylase which

has biotin as a prosthetic group, a common feature in CO2-binding
enzymes.

• One molecule of ATP is hydrolyzed in the reaction, which is irreversible.

• The elongation steps of fatty acid synthesis all involve intermediates

linked to the terminal sulfhydryl group of the phosphopantetheine
reactive unit in ACP; phosphopantetheine is also the reactive unit in CoA.
• Therefore, the next steps are the formation of acetyl-ACP and
malonyl-ACP by the enzymes acetyl transacylase and
malonyl transacylase, respectively .

• For the synthesis of fatty acids with an odd number of carbon

atoms the three-carbon propionyl- ACP is the starting point
instead of malonyl-ACP.)
• The elongation cycle of fatty acid synthesis has four stages for
each round of synthesis . For the first round of synthesis these
are:
• 1. Condensation of acetyl-ACP and malonyl-ACP to form
acetoacetyl-ACP, releasing free ACP and CO2 (catalyzed by
acyl-malonyl-ACP condensing enzyme).
• 2. Reduction of acetoacetyl-ACP to form D-3-hydroxybutyryl-
ACP, using NADPH as reductant (catalyzed by -ketoacyl-ACP
reductase).
• 3. Dehydration of D-3-hydroxybutyryl-ACP to produce crotonyl-
ACP (catalyzed by 3-hydroxyacyl-ACP dehydratase).
• 4. Reduction of crotonyl-ACP by a second NADPH molecule to
give butyryl- ACP (catalyzed by enoyl-ACP reductase).
Fatty Acid Regulation
• Acetyl CoA carboxylase plays an essential role in the regulation
of fatty acid synthesis and degradation.
• Acetyl CoA carboxylase catalyzes the committed step of fatty
acid biosynthesis.
• Fatty acid oxidation is regulated largely by fatty acids in the
blood, which is controlled by the hydrolysis rate of
triacylglycerols in the adipose tissue by hormone-sensitive
triacylglycerol lipase.
• AMPK also regulates fatty acid oxidation
Diseases associated with fatty acid
catabolism

• Mitochondrial fatty acid β-oxidation disorders

(FAODs)