4rd week, 2nd Semester

College of Medicine
Fatty Acid and Triacylglycerol Al Imam Mohammad Ibn Saud Islamic University
Metabolism II Female Section
Dr Gihan AlHusseini Gawish
Lippincott’s Illustrated Reviews: Biochemistry : 5 th Edition

OBJECTIVES
1) Describe the mechanism for activation and transport of fatty acids into mitochondria for

catabolism✓.

2) Outline the sequence of reactions involved in oxidation of fatty acids in mitochondria ✓.

3) Describe the general features of pathways for oxidation of unsaturated odd-chain and branched-

chain fatty acids ✓.

4) Explain the mechanism for the formation of ketone bodies and identify the physiological and

pathological roles of those molecules ✓. 1

Dr Gihan AlHusseini Gawish 2
  Fatty acids stored in adipose tissue, in the form of neutral TAG, serve as
the body’s major fuel storage reserve.
 TAGs provide concentrated stores of metabolic energy BECAUSE they are
highly reduced and largely anhydrous.
 The yield from the complete oxidation of fatty acids to CO2 and H2O is 9
kcal/g fat (as compared to 4 kcal/g protein or carbohydrate
 Fatty acids are also supplied to tissues by lipoproteins.

MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

A. Release of fatty acids from TAG

B. β-Oxidation of fatty acids

C. α-Oxidation of fatty acids

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Dr Gihan Gawish
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

A.Release of fatty acids from TAG

 Hydrolytic release
 Hormone-sensitive lipase removes a fatty acid from
C1and/or C3 of the TAG.
 Additional lipases specific for DAG / MAG remove the
remaining FAs.
1. Activation of hormone-sensitive lipase (HSL):

 Activated when phosphorylated by 3',5'cAMP

dependent protein kinase.

 cAMP–dpk produced in the adipocyte when one of

several hormones (epinephrine or glucagon) binds
to receptors on the cell membrane, and activates
adenylyl cyclase.

 High plasma levels of insulin and glucose HSL 4

Hormonal regulation of triacylglycerol
is dephosphorylated,
Dr Gihan Gawish and becomes inactive. degradation in the adipocyte.
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

A.Release of fatty acids from TAG

2. Fate of glycerol:

 The glycerol released during TAG degradation cannot

be metabolized by adipocytes ?

(They apparently lack glycerol kinase).

 Glycerol is transported through the blood to the liver to

phosphorylated.

 Glycerol phosphate used to TAG in the liver OR

converted to DHAP by reversal of the glycerol

phosphate dehydrogenase.
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Hormonal regulation of triacylglycerol
Dr Gihan Gawish degradation in the adipocyte.
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

A.Release of fatty acids from TAG

3. Fate of fatty acids:

FFAs move bind to plasma albumin (plasma membrane)

Transported to the tissues, enter cells, get activated to their CoA derivatives, and
are oxidized for energy.
(FFA) cannot be used for fuel by erythrocytes? Brain, too??
Over 50% of the fatty acids released from
adipose TAG are reesterified to glycerol 3 phosphate.
White adipose does not express glycerol kinase, and the phosphorylated glycerol
is produced by glyceroneogenesis, an incomplete version of gluconeogenesis
pyruvate to PEP to OAA to DHAP to glycerol 3-phosphate.
The process reduces plasma FFA, molecules associated with insulin resistance in
Dr Gihan Gawish 6
type 2 diabetes and obesity.
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids mitochondrial pathway

two-carbon fragments removed from the carboxyl end of the fatty acyl CoA producing
acetyl CoA + NADH + FADH2.

1. Transport of long-chain fatty acids (LCFA)

Carnitine shuttle. [Note: Long-chain fatty acyl CoA synthetase is in the outer mitochondrial
membrane; active site faces the cytosol.] 7
Dr Gihan Gawish
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids mitochondrial pathway

1. Transport of long-chain fatty acids (LCFA) Carnitine shuttle

a. Steps in LCFA translocation:

 First: Acyl group is transferred from CoA to carnitine by carnitine
palmitoyltransferase I
 (CPT-I)—an enzyme of the outer mitochondrial membrane.
 CPT-I is also known as CAT-I for carnitine acyltransferase
 This reaction forms acyl carnitine, and regenerates free CoA.
 Second: Acylcarnitine is transported into the mitochondrial matrix in exchange
for free carnitine by carnitine–acylcarnitine translocase.
 Carnitine palmitoyltransferase II (CPT-II, or CAT-II)—an enzyme of the inner
mitochondrial membrane—catalyzes theGawish
Dr Gihan transfer of the acyl group from carnitine
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to CoA in the mitochondrial matrix, thus regenerating free carnitine.

 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids

1. Transport of long-chain fatty acids (LCFA)

b. Inhibitor of the carnitine shuttle:

 Malonyl CoA inhibits CPT-I, preventing the entry of long-
chain acyl groups M Matrix.
 New palmitate transferred into the mitochondria and
degraded.
 Muscle not synthesize fatty acids, contains the
mitochondrial isoform of acetyl CoA carboxylase (ACC2 ),
allowing muscle to regulate β-oxidation.
 FA oxidation is regulated by acetyl CoA to CoA ratio: as the
ratio increases, the CoArequiring thiolase reaction
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decreases.
Enzymes involved in the β-oxidation of fatty acyl CoA. [Note: Enoyl CoA hydratase requires a trans
double bond between carbon 2 and carbon 3
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids

1. Transport of long-chain fatty acids (LCFA)

c. Sources of carnitine:
Carnitine can be obtained from the diet

Carnitine can also be synthesized from the amino acids lysine and methionine by
an enzymatic pathway found in the liver and kidney

 Not in skeletal or heart muscle, These tissues are totally dependent on uptake of
carnitine provided by endogenous synthesis or the diet, and distributed by the
blood.

Dr Gihan Gawish 10
Skeletal muscle contains about 97% of all carnitine in the body
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids

1. Transport of long-chain fatty acids (LCFA)

d. Carnitine deficiencies:
1) In patients with liver disease causing decreased synthesis of Carnitine
2) In individuals suffering from malnutrition or vegetarian diets
3) In those with an increased requirement for carnitine as a result of, for example,
pregnancy, severe infections, burns, or trauma;
4) in those undergoing hemodialysis, which removes carnitine from the blood.

Genetic CPT-I deficiency affects the liver, where an inability to use

LCFA for fuel greatly impairs that tissue’s ability to synthesize glucose
during a fast.
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Dr Gihan Gawish
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids

2. Entry of short- and medium-chain fatty acids into the mitochondria:)

 FAs < 12 C cross IMM without Carnitine /CPT system.

 Then activated to their CoA derivatives by matrix enzymes, and are oxidized.

 Medium-chain FAs are plentiful in human milk.

Because their oxidation is not dependent on CPT-I, it is not subject to inhibition by

malonyl CoA.

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Dr Gihan Gawish
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids

3. Reactions of β-oxidation:

The first cycle of β-oxidation consists of a sequence of four

RXs involving
the β-carbon (C 3)

shortening the FAs by 2C.

steps include:
1.oxidation FADH2
2.hydration
3.oxidation NADH
4.thiolytic cleavage releases an acetyl CoA.
Each step is catalyzed by enzymes with chain-length
specificity.
These steps are repeated for saturated FAs of
carbon chains (n/2) – 1 times= (odd) 13
The final thiolytic cleavage produces two acetyl groups
Dr Gihan Gawish
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids

4. Energy yield from fatty acid oxidation:

β-oxidation pathway yield high Energy.

Ex: Oxidation of a Palmitoyl CoA to CO2 and H2O

8 acetyl CoA+ 7NADH+ 7FADH2 = 131 ATP

however, activation of FA requires 2 ATP.

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Dr Gihan Gawish
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids

5. Medium-chain fatty acyl CoA dehydrogenase (MCAD) deficiency
Four fatty acyl CoA dehydrogenase species mitochondria

Each with a specificity for either short-, medium-, long-, or very-long-chain fatty acids.

MCAD deficiency, an autosomal recessive disorder, is one of the most common inborn
errors of metabolism, and the most common inborn error of fatty acid oxidation

1:14,000 births worldwide, with a higher incidence in Northern Europeans.

It results in decreased ability to oxidize fatty acids with six to ten carbons (these
accumulate and can be measured in urine), and severe hypoglycemia (because the tissues
must increase their reliance on glucose).

Treatment includes avoidance of fasting.

MCAD deficiency has been identified as the cause of some cases originally reported as
sudden infant death syndrome (SIDS) or Reye syndrome.
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Dr Gihan Gawish
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids

6. Oxidation of fatty acids with an odd number of carbons:

β-oxidation of a saturated FA with an odd C# proceeds by the

same reaction steps as that of FAs with an even C#, until the
final three carbons are reached.
This compound, propionyl CoA, is metabolized by a three-step
pathway

a. Synthesis of D-methylmalonyl CoA

b. Formation of L-methylmalonyl CoA

c. Synthesis of succinyl CoA

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Dr Gihan Gawish
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids

7. Oxidation of unsaturated fatty acids:

 The oxidation of unsaturated FAs provides less energy than that of saturated fatty acids

 Because unsaturated FAs are less highly reduced

 Oxidation of monounsaturated fatty acids, such as 18:1(9) (oleic acid) requires one

additional enzyme, 3,2-enoyl CoA isomerase

 Oxidation of polyunsaturated fatty acids, such as 18:2(9,12) (linoleic acid), requires an

NADPH-dependent 2,4-dienoyl CoA reductase in addition to the isomerase.

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Dr Gihan Gawish
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

B. β-Oxidation of fatty acids

8. β-Oxidation in the peroxisome:

 VLCFA(22 C or longer) undergo a preliminary β-oxidation in peroxisomes.

 The initial dehydrogenation in peroxisomes is catalyzed by an FAD-containing acyl CoA
oxidase.
 The FADH2 produced is oxidized by molecular oxygen, which is reduced to H2O2; (no
ATP is generated by this step).
 The H2O2 is reduced to H2O by catalase

Genetic defects either in the ability to target matrix proteins to peroxisomes (resulting in
Zellweger syndrome —a peroxisomal biogenesis disorder) or in the ability to transport
VLCFA across the peroxisomal membrane (resulting in X-linked
adrenoleukodystrophy), lead to accumulation of VLCFA in the blood and tissues.18
Dr Gihan Gawish
 MOBILIZATION OF STORED FATS AND OXIDATION OF FATTY ACIDS

C. α-Oxidation of fatty acids

Branched-chain, 20 carbon fatty acid, phytanic acid
It is not substrate for acyl CoA dehydrogenase because of the
methyl group on its β carbon.
it is hydroxylated at the α-carbon by phytanoyl CoA α-
hydroxylase (PhyH), carbon 1 is released as CO2, and the
product, 19 carbon pristanic acid, is activated to its CoA
derivative and undergoes β-oxidation.

Refsum disease
Rare autosomal recessive disorder caused by a deficiency of peroxisomal PhyH.
Results in the accumulation of phytanic acid in the plasma and tissues.
The symptoms are primarily neurologic
treatment : dietary restriction to halt disease progression. 19
Dr Gihan Gawish
 KETONE BODIES: AN ALTERNATE FUEL FOR CELLS
 Liver mitochondria have the capacity to convert acetyl CoA derived from
FAs oxidn into ketone bodies.
(acetoacetate, β-hydroxybutyrate, and acetone)

 They are transported in the blood to the peripheral tissues.

 They can be reconverted to acetyl CoA, which can be oxidized by the TCA
cycle.

 Ketone bodies are important sources of energy for the peripheral tissues
BECAUSE they are soluble in aqueous solution & not need to be
incorporated into lipoproteins or carried by albumin

 They are produced in the liver during periods when the amount of acetyl
CoA present exceeds the oxidative capacity of the liver

 They are used in proportion to their concentration in the blood by

extrahepatic tissues, such as the skeletal and cardiac muscle,renal cortex
and even brain

 This is particularly important during prolonged periods of fasting .

 Disorders of fatty acid oxidation present with the general picture of

hypoketosis (due to decreased availability of acetyl CoA) and hypoglycemia 20
(due to increased reliance on glucose for energy.
Dr Gihan Gawish
Ketone body synthesis in the liver and use in peripheral tissues

A. Synthesis of ketone bodies by the liver: Ketogenesis

 During a fast, the liver is flooded with fatty acids mobilized from adipose tissue.

 The resulting elevated hepatic acetyl CoA produced primarily by fatty acid degradation inhibits pyruvate
dehydrogenase and activates pyruvate carboxylase .

 The OAA thus produced is used by the liver for gluconeogenesis rather than for the TCA cycle.

 Therefore, acetyl CoA is channeled into ketone body synthesis.

 Fatty acid oxidation decreases the NAD+ to NADH ratio, and the rise in NADH shifts OAA to malate
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 This pushes acetyl CoA away from gluconeogenesis and into ketogenesis
 KETONE BODIES: AN ALTERNATE FUEL FOR CELLS

A. Synthesis of ketone bodies by the liver: Ketogenesis

1. Synthesis of 3-hydroxy-3-methylglutaryl 2. Synthesis of the ketone bodies:
(HMG) CoA:
 HMG CoA is cleaved to produce acetoacetate
 Formation of acetoacetyl CoA, occurs by and acetyl CoA
reversal of the thiolase reaction of fatty acid  Acetoacetate can be reduced to form 3-
oxidation. hydroxybutyrate with NADH as the hydrogen
donor.
 Mitochondrial HMG CoA synthase combines  Acetoacetate can also spontaneously
a third molecule of acetyl CoA with decarboxylate in the blood to form acetone
acetoacetyl CoA to produce HMG CoA.  It is a biologically nonmetabolized compound
that can be released in the breath.
 HMG CoA is a precursor of cholesterol  The equilibrium between acetoacetate and 3-
hydroxybutyrate is determined by the
 These pathways are separated by location in, NAD+/NADH ratio.
and conditions of, the cell.  Because this ratio is low during fatty acid
oxidation, 3-hydroxy butyrate synthesis is
 HMG CoA synthase is the rate-limiting step favored.
in the synthesis of ketone bodies, and is  The generation of free CoA during keto -
present in significant quantities only in the genesis allows fatty acid oxidation to continue.
liver. 22
Dr Gihan Gawish
 KETONE BODIES: AN ALTERNATE FUEL FOR CELLS

B. Use of ketone bodies by the peripheral tissues:ketolysis

 liver ketone bodiesbecomes much more significant during fasting when ketone bodies are needed to provide energy
to the peripheral tissues.

 3-Hydroxybutyrate is oxidized to acetoacetate by 3-HB dehydrogenase, producing NADH

 Aceto acetate is then provided with a CoA molecule taken from succinyl CoA by succinyl CoA:acetoacetate CoA
transferase (thiophorase).

 This reaction is reversible, but the product, acetoacetyl CoA, is actively removed by its conversion to two acetyl
CoA.

 Extrahepatic tissues, including the brain but excluding cells lacking mitochondria (for example, red blood cells),
efficiently oxidize acetoacetate and 3-HB

 Although the liver actively produces ketone bodies, it lacks thiophorase and, therefore, is unable to use ketone
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bodies as fuel.
Dr Gihan Gawish
 KETONE BODIES: AN ALTERNATE FUEL FOR CELLS

C. Excessive production of ketone bodies in diabetes: ketoacidosis

When the rate of formation of ketone bodies is greater
mellitus
than the rate of their use, their levels begin to rise in
(ketonemia) and, (ketonuria), type 1 diabetes mellitus.

In diabetic individuals with severe ketosis, urinary

excretion of the ketone bodies > 5,000 mg/24 hr, and the
blood concentration may reach 90 mg/dl (Vs >3 mg/dl in
normal individuals).

A frequent symptom of diabetic ketoacidosis is a fruity

odor on the breath,(increased production of acetone).

An elevation of the ketone body concentration in the blood

results in acidemia.The carboxyl group of a ketone body

Also, excretion of glucose and ketone bodies in the urine

results in dehydration of the body.
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Ketoacidosis may also be seen in cases of fasting
Dr Gihan Gawish
APPROACHED OBJECTIVES
1) Describe the mechanism for activation and transport of fatty acids into mitochondria for
catabolism✓.
2) Outline the sequence of reactions involved in oxidation of fatty acids in mitochondria ✓.
3) Describe the general features of pathways for oxidation of unsaturated odd-chain and
branched-chain fatty acids ✓.
4) Explain the mechanism for the formation of ketone bodies and identify the physiological and
pathological roles of those molecules ✓. Dr Gihan Gawish 25