Metabolism of Fatty Acids

(Beta Oxidation, synthesis of ketones,

Synthesis of Fatty Aids)
Dr. Anwar Ullah
 Digestion, Mobilization, and Transport of Fats
• Cells can obtain fatty acid fuels from three sources:
1. fats consumed in the diet.

2. fats stored in cells as lipid droplets.

3. fats synthesized in one organ for export to another.

• Some species use all three sources under various circumstances, others

use one or two.

• Vertebrates, for example, obtain fats in the diet, mobilize fats stored

in specialized tissue (adipose tissue, consisting of cells called

adipocytes), and, in the liver, convert excess dietary carbohydrates to

fats for export to other tissues.

 Digestion, Mobilization, and Transport of Fats

• On average, 40% or more of the daily energy requirement of

humans in highly industrialized countries is supplied by dietary
triacylglycerols (although most nutritional guidelines
recommend no more than 30% of daily caloric intake from fats).
• Triacylglycerols provide more than half the energy
requirements of some organs, particularly the liver, heart,
and resting skeletal muscle.
• Stored triacylglycerols are virtually the sole source of energy in
hibernating animals and migrating birds
 Dietary Fats Are Absorbed in the Small Intestine
• In vertebrates, before ingested triacylglycerols can be absorbed
through the intestinal wall they must be converted from insoluble
macroscopic fat particles to finely dispersed microscopic micelles.
• This solubilization is carried out by bile salts, such as taurocholic
acid which are synthesized from cholesterol in the liver, stored in
the gallbladder, and released into the small intestine after ingestion
of a fatty meal.
• Bile salts are amphipathic compounds that act as biological
detergents, converting dietary fats into mixed micelles of bile salts
and triacylglycerols
Lipid Digestion
 Stages of fatty acid oxidation

(1) Activation of fatty acids takes place on the outer

mitochondrial membrane
(2) Transport into the mitochondria

(3) Degradation to two-carbon fragments (as acetyl CoA) in

the mitochondrial matrix (b-oxidation pathway)
 (1) Activation of Fatty Acids
• Fatty acids are converted to CoA thioesters by acyl-CoA
synthetase (ATP dependent)
• The PPi released is hydrolyzed by a pyrophosphatase to 2 Pi
• Two phosphoanhydride bonds (two ATP equivalents) are
consumed to activate one fatty acid to a thioester
(2) Transport of Fatty Acyl CoA into Mitochondria

• The carnitine shuttle system.

• Fatty acyl CoA is first converted
to acylcarnitine (enzyme
carnitine acyltransferase I
(bound to the outer
mitochondrial membrane).
• Acylcarnitine enters the
mitochondria by a translocase.
• The acyl group is transferred
back to CoA (enzyme -
carnitine acyltransferase II).
• Carnitine
shuttle system
• Path of acyl
group in red
(3) The reaction of Beta Oxidation
 (3) The Reactions of b oxidation
• The b-oxidation pathway 1. Oxidation of acyl CoA by an
(b-carbon atom (C3) is acyl CoA dehydrogenase to give an
oxidized) degrades fatty enoyl CoA
acids two carbons at a Coenzyme - FAD
time



2. Hydration of the double
bond between C-2 and C-3 by
enoyl CoA hydratase with the
3-hydroxyacyl CoA (b-
hydroxyacyl CoA) formation
3. Oxidation of 3-hydroxyacyl
CoA to 3-ketoacyl CoA by 3-
hydroxyacyl CoA dehydrogenase
Coenzyme – NAD+
4. Cleavage of 3-ketoacyl
CoA by the thiol group of
a second molecule of CoA
with the formation of
acetyl CoA and an acyl
CoA shortened by two
carbon atoms.
Enzyme - b-ketothiolase.
The shortened acyl CoA
then undergoes another
cycle of oxidation

The number of cycles:

n/2-1, where n – the
number of carbon atoms
 b-Oxidation of
Fatty acyl CoA
saturated fatty acids
• One round of b oxidation: 4 enzyme steps produce acetyl CoA
from fatty acyl CoA
• Each round generates one molecule each of:
FADH2
NADH
Acetyl CoA
Fatty acyl CoA (2 carbons shorter each round)

Fates of the products of b-oxidation:- NADH and FADH2 - are used

in ETC - acetyl CoA - enters the citric acid cycle - acyl CoA –
undergoes the next cycle of oxidation
ATP Generation from Fatty Acid Oxidation
Net yield of ATP per one oxidized palmitate
Palmitate (C15H31COOH) - 7 cycles – n/2-1

The balanced equation for oxidizing one palmitoyl CoA by

seven cycles of b oxidation
Palmitoyl CoA + 7 HS-CoA + 7 FAD+ + 7 NAD+ + 7 H2O
8 Acetyl CoA + 7FADH2 + 7 NADH + 7 H+
ATP generated
8 acetyl CoA 10x8=80
7 FADH2 7x1.5=10.5
7 NADH 7x2.5=17.5
108 ATP
ATP expended to activate palmitate -2
Net yield: 106 ATP
 B-oxidation of odd-chain fatty acids

• Odd-chain fatty acids occur in

bacteria and microorganisms
• Final cleavage product is
propionyl CoA rather than acetyl
CoA
• Three enzymes convert propionyl
CoA to succinyl CoA (citric acid
cycle intermediate)
Propionyl CoA Is Converted into Succinyl CoA
1. Propionyl CoA is carboxylated to yield the D isomer of
methylmalonyl CoA.
The hydrolysis of an ATP is required.
Enzyme: propionyl CoA carboxylase
Coenzyme: biotin
 2. The D isomer of methylmalonyl CoA is racemized to the L
isomer
Enzyme: methylmalonyl-CoA racemase

3. L isomer of methylmalonyl CoA is converted into succinyl CoA

by an intramolecular rearrangement
Enzyme: methylmalonyl CoA mutase
Coenzyme: vitamin B12 (cobalamin)
β-Oxidation of Polyunsaturated Fatty Acid
 Ketone Bodies
The entry of acetyl CoA into the citric acid cycle depends
on the availability of oxaloacetate.
The concentration of oxaloacetate is lowered if
carbohydrate is unavailable (starvation) or improperly
utilized (diabetes).

Oxaloacetate is
normally formed from
pyruvate by pyruvate
carboxylase
(anaplerotic reaction).

Fats burn in the flame

of carbohydrates.
In fasting or diabetes the gluconeogenesis is activated and
oxaloacetate is consumed in this pathway.
Fatty acids are oxidized producing excess of acetyl CoA which is
converted to ketone bodies:
B-Hydroxybutyrate
Acetoacetate
Acetone

Ketone bodies are synthesized in

liver mitochondria and exported to
different organs.

Ketone bodies are fuel molecules (can

fuel brain and other cells during
starvation)
 A. Synthesis of ketone bodies
Two molecules of acetyl
CoA condense to form
acetoacetyl CoA.
Enzyme – thiolase.

Acetoacetyl CoA reacts

with acetyl CoA and
water to give 3-hydroxy-
3 methylglutaryl CoA
(HMG-CoA) and CoA.
Enzyme: HMG-CoA
synthase
3-Hydroxy-3-
methylglutaryl CoA is then
cleaved to acetyl CoA and
acetoacetate.
Enzyme: HMG-CoA lyase.

3-Hydroxybutyrate is formed
by the reduction of acetoacetate by
3-hydroxybutyrate dehydrogenase.
Acetoacetate also undergoes a
slow, spontaneous decarboxylation
to acetone.
The odor of acetone may be
detected in the breath of a person
who has a high level of
acetoacetate in the blood.
 B. Ketone bodies are a major fuel in some
tissues
1. Ketone bodies diffuse from the liver mitochondria into the
blood and are transported to peripheral tissues.
2. Ketone bodies are important molecules in energy metabolism.
3. Heart muscle and the renal cortex use acetoacetate in
preference to glucose in physiological conditions.
4. The brain adapts to the utilization of acetoacetate during
starvation and diabetes.
3-Hydroxybutyrate is oxidized to produce acetoacetate as
well as NADH for use in oxidative phosphorylation.

3-
hydroxybutyrate
dehydrogenase
Acetoacetate is activated by the
transfer of CoA from succinyl
CoA in a reaction catalyzed by a
specific CoA transferase.
Acetoacetyl CoA is cleaved by
thiolase to yield two molecules of
acetyl CoA (enter the citric acid
cycle).
CoA transferase is present in all
tissues except liver.

Ketone bodies are a water-

soluble, transportable form of
acetyl units
 Ketosis
The absence of insulin in diabetes mellitus

 liver cannot absorb glucose  activation of fatty acid

 inhibition of glycolysis mobilization by adipose
 activation of gluconeogenesis tissue

 deficit of oxaloacetate

 large amounts of acetyl CoA which can not

be utilized in Krebs cycle

 large amounts of ketone bodies (moderately strong acids)

 severe acidosis (ketosis)

Impairment of the tissue function, most importantly in the central

nervous system
 Fatty Acid Synthesis

• Occurs mainly in liver and adipocytes, in mammary glands

during lactation

• Occurs in cytoplasm

• FA synthesis and degradation occur by two completely

separate pathways

• When glucose is plentiful, large amounts of acetyl CoA are

produced by glycolysis and can be used for fatty acid
synthesis
Three stages of fatty acid synthesis

A. Transport of acetyl CoA into cytosol

B. Carboxylation of acetyl CoA

C. Assembly of fatty acid chain

A. Transport of Acetyl CoA to the Cytosol

• Acetyl CoA from catabolism of carbohydrates and amino

acids is exported from mitochondria via the citrate
transport system

• Cytosolic NADH also converted to NADPH

• Two molecules of ATP are expended for each round of this

cyclic pathway
Citrate transport
system
 Sources of NADPH for Fatty Acid Synthesis
1. One molecule of NADPH is generated for each molecule
of acetyl CoA that is transferred from mitochondria to the
cytosol (malic enzyme).

2. NADPH molecules come from the pentose phosphate

pathway.
 B. Carboxylation of Acetyl CoA
Enzyme: acetyl CoA carboxylase Prosthetic group - biotin

A carboxybiotin intermediate is formed.

ATP is hydrolyzed.
The CO2 group in carboxybiotin is transferred to acetyl CoA
to form malonyl CoA.
Acetyl CoA carboxylase is the regulatory enzyme.
 C. The Reactions of Fatty Acid Synthesis
• Five separate stages:
(1) Loading of precursors via thioester derivatives
(2) Condensation of the precursors
(3) Reduction
(4) Dehydration
(5) Reduction
During the fatty acid synthesis all intermediates are linked to
the protein called acyl carrier protein (ACP-SH), which is the
component of fatty acyl synthase complex.

The pantothenic acid is a

component of ACP.
Intermediates in the
biosynthetic pathway are
attached to the sulfhydryl
terminus of
phosphopantotheine group.
The elongation phase of fatty acid synthesis starts with the
formation of acetyl ACP and malonyl ACP.
Acetyl transacylase and malonyl transacylase catalyze
these reactions.
Acetyl CoA + ACP  acetyl ACP + CoA
Malonyl CoA + ACP  malonyl ACP + CoA
Condensation reaction.
Acetyl ACP and malonyl ACP
react to form acetoacetyl ACP.
Enzyme -acyl-malonyl ACP
condensing enzyme.
Reduction.
Acetoacetyl ACP is reduced to
D-3-hydroxybutyryl ACP.
NADPH is the reducing agent
Enzyme: -ketoacyl ACP
reductase
Dehydration.
D-3-hydroxybutyryl ACP is
dehydrated to form crotonyl
ACP
(trans-2-enoyl ACP).
Enzyme: 3-hydroxyacyl ACP
dehydratase
Reduction.
The final step in the cycle
reduces crotonyl ACP to
butyryl ACP.
NADPH is reductant.
Enzyme - enoyl ACP reductase.
This is the end of first
elongation cycle (first round).
In the second round butyryl ACP
condenses with malonyl ACP to
form a C6--ketoacyl ACP.
Reduction, dehydration, and a
second reduction convert the C6-

-ketoacyl ACP into a C6-acyl

ACP, which is ready for a third
round of elongation.
 Final reaction of FA synthesis
• Rounds of synthesis continue until a
C16 palmitoyl group is formed
• Palmitoyl-ACP is hydrolyzed by a thioesterase

Overall reaction of palmitate synthesis from acetyl CoA and

malonyl CoA

Acetyl CoA + 7 Malonyl CoA + 14 NADPH + 14 H+

Palmitate + 7 CO2 + 14 NADP+ + 8 HS-CoA + 6 H2O
 Fatty Acid Elongation and Desaturation
The common product of fatty acid synthesis is palmitate (16:0).

Cells contain longer fatty acids and unsaturated fatty acids they
are synthesized in the endoplasmic reticulum.

The reactions of elongation are similar to the ones seen with

fatty acid synthase (new carbons are added in the form of
malonyl CoA).

For the formation of unsaturated fatty acids there are various

desaturases catalizing the formation of double bonds.
 The control of fatty acid metabolism
Acetyl CoA carboxylase plays an essential role in
regulating fatty acid synthesis and degradation.
The carboxylase is controlled by hormones:
 glucagon,
 epinephrine, and
 insulin.
Another regulatory factors:
 citrate,
 palmitoyl CoA, and
 AMP
 Global Regulation
is carried out by means of reversible phosphorylation
Acetyl CoA carboxylase is switched off by phosphorylation and
activated by dephosphorylation
Insulin stimulates fatty acid synthesis causing dephosphorylation of
carboxylase.
Glucagon and epinephrine have the reverse effect (keep the
carboxylase in the inactive phosphorylated state).

Protein kinase is activated

by AMP and inhibited by
ATP.
Carboxylase is
inactivated when the
energy charge is low.
 Local Regulation
Acetyl CoA carboxylase is allosterically stimulated by citrate.
The level of citrate is high when both acetyl CoA and ATP are
abundant (isocitrate dehydrogenase is inhibited by ATP).
Palmitoyl CoA inhibits carboxylase.
 Response to Diet
Fed state:
• Insulin level is increased
• Inhibits hydrolysis of stored TGs
• Stimulates formation of malonyl CoA, which inhibits carnitine
acyltransferase I
• FA remain in cytosol (FA oxidation enzymes are in the
mitochondria)
Starvation:
• Epinephrine and glucagon are produced and stimulate adipose
cell lipase and the level of free fatty acids rises
• Inactivate carboxylase, so decrease formation of malonyl CoA
(lead to increased transport of FA into mitochondria and activate
the b-oxidation pathway)
Thank You