Lecture 17

Lipid Metabolism
Metabolism of Fatty acids - I

Dr. S. Annie Jeyachristy

Lecturer in Biochemistry
Faculty of Medicine
AIMST University
 OBJECTIVES

To provide an overview of
Metabolism of lipids
Synthesis of fatty acids and its regulation
Synthesis of triacylglycerol in adipose tissue
Describe the formation and utilization of ketone bodies.
Acetyl Co A as key Intermediate between fat and
carbohydrate metabolism
Carbohydrates can be converted to
fatty acids but not vice-versa
except propionyl CoA
Pyruvate dehydrogenase (PDH)
virtually irreversible → pyruvate
cannot be formed from acetyl CoA
(or)
Acetyl coA cannot be converted
back to pyruvate in animals.
DE NOVO SYNTHESIS OF FATTY ACIDS (FA)
Referred to as ‘Lynen’s spiral’
Extramitochondrial system – Cytosol
Liver, adipose tissue, kidney, brain, and in mammary glands during
lactation
Cofactors – NADPH, ATP, Mn2+, biotin, HCO3- (Source of CO2)
Immediate substrate – Acetyl CoA
End product – free palmitate
Three stages of FA synthesis

A. Transport of acetyl CoA into cytosol

B. Carboxylation of acetyl CoA to form malonyl CoA
C. Assembly of fatty acid chain
Transport of acetyl CoA into cytosol

Acetyl CoA is formed inside the mitochondria, from catabolism of

carbohydrates and amino acids, like all CoASH derivatives cannot
penetrate the mitochondrial membrane, and is exported from
mitochondria via the citrate transport system
Acetyl groups are delivered to the cytoplasm as citrate.
Citrate is transported from mitochondria by a tricarboxylic acid
transporter.
In the cytoplasm, citrate is cleaved to oxaloacetate and acetyl CoA in
the cytoplasm.
The enzyme is ATP citrate lyase
The oxaloacetate can return to the mitochondria as malate and
pyruvate.
Fatty acid synthase complex
Multienzyme complex – dimer with identical subunits
Seven separate polypeptides, that are tightly associated to form the complex is
organized into 3 domains.
Seven polypeptides are made up of six enzymes and one acyl carrier protein, ACP
1st domain or condensing unit – initial substrate binding site.
beta-ketoacyl synthase or condensing enzyme (CE);
acetyl transferase (AT), and
malonyl transacetylase (MT)
2nd domain or reduction unit –
Dehydratase (DH)
enoyl reductase (ER)
beta-keto acyl reductase (KR) and
acyl carrier protein (ACP)
3rd domain or releasing unit – involved in the release of synthesized palmitate.
thio-esterase (TE) or deacylase.
 ACP is a polypeptide chain having a phospho pantotheine group, to which the acyl
groups are attached in thioester linkage. ACP acts like CoA carrying fatty acyl
groups.

• In prokaryotes, reactions of FA synthesis carried out by separate enzymes.

• In eukaryotes, enzymes present in a single polypeptide chain, multifunctional
enzyme complex called fatty acid synthase (FAS).
Carboxylation of acetyl CoA to form malonyl CoA – committed step

Enzyme: acetyl CoA carboxylase - the regulatory enzyme

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.
Allosteric regulation. positive – citrate; negative – palmitoyl CoA
Formation of acetyl- and malonyl- acyl carrier protein (ACP)

The acetyl transacylase catalyses the transfer of the acetyl group to the cysteinyl SH group of
CE of the one monomer of FAS complex
One molecule of acetyl CoA and one molecule of malonyl CoA bind to the enzyme complex.
Malonyl transacylase transfers the malonyl group to the SH group of the ACP of the other
monomer of the enzyme
 Cycle of reactions involve
 Condensation of acetyl–ACP
and malonyl–ACP with release
of CO2.
 Reduction with NADPH.
 Dehydration to yield a trans
double bond.
 Reduction with NADPH.

First cycle yields butyryl–ACP (C4).

Condensing enzyme – β-ketoacyl ACP synthase

  In the second round of fatty acid
synthesis, butyryl ACP condenses
with malonyl ACP to form ß-
ketoacyl-ACP.
 Reduction, dehydration and a second
reduction convert the Ketoacyl ACP
into acyl ACP.
 The elongation cycles continue until
C16 acyl ACP
 Then it is hydrolyzed to yield
palmitate and ACP

Thioesterase
The end point is palmitic acid (C16) in liver and adipose tissue.
In lactating mammary glands, the end products are Capric (C10) and
Lauric acid (C12) – medium chain fatty acids
Cow’s milk contains odd numbered fatty acids

Co-enzymes of FAS

NADPH
1. Pentose phosphate pathway – HMP shunt pathway
2. Malic enzyme
 REGULATION OF FATTY ACID BIOSYNTHESIS
1. Availability of substrates.
 The synthesis of fatty acid is maximal when carbohydrate is abundant and the
level of fatty acid is low.
 Availability of citrate in the cytoplasm – short-term effect
2. Acetyl coA carboxylase
 rate limiting enzyme of fatty acid biosynthesis.
 Citrate activates this enzyme. Citrate is high when acetyl CoA and ATP are
abundant.
 Feedback inhibition - Palmitoyl CoA
 Covalent modification - Phosphorylation (inactivates) and dephosphorylation
(activates) of acetyl CoA carboxylase
3. Insulin-enhance lipogenesis (PDH, acetyl CoA carboxylase and glycerol
phosphate acyl transferase); also depresses hormone sensitive lipase.
4. Glucagon and epinephrine inhibits lipogenesis – phosphorlyating acetyl CoA
carboxylase
 Fatty Acid Modification
Synthesis of odd-chain fatty acids
Propionyl –ACP (3-C) used as primer – occurs particularly in
ruminants. Condensation of propionyl-ACP and malonyl-ACP leads
to formation of 5C fatty acid.
Further condensation sequentially with malonyl-ACP leads to
extension of chain of odd C atoms

Chain Elongation
In animals, palmitate (C16) is the principal product of FAS
Palmitate is the precursor for the formation of other long-chain fatty
acids
Elongation of fatty acids (beyond C16) occur both in the mitochondria
as well as endoplasmic reticulum, especially in the latter known as the
microsomal system
 •
Desaturation of Fatty Acids

• Palmitoyl CoA + NADPH + H+ + O2 → Palmitoeyl CoA + NADP+ + 2H2O

Involves microsomal system called fatty acyl-CoA desaturase which introduces

the double bond into the fatty acid chain
Mammals cannot introduce double bonds beyond ∆9 in fatty acids
Unsaturated fatty acids like linoleic acid (ω6, 18:2, ∆9,12) and linolenic acid (ω3,
18:3, ∆9,12,15), are the only two known essential fatty acids in many animals,
including humans; provided in the diet.
SYNTHESIS OF TRIACYLGLYCEROL (TAG)
Liver and adipose tissue are the
major site
Fatty acids are converted to
Glycerol triacylglycerols (TAG) for storage
phosphate especially in adipose tissue and, in
Pathway liver secreted as VLDL and is
transported
Stored TAG in dynamic state –
Monoacylglycerol continuously broken down and
Pathway synthesized
Biosynthesis of TAG – 2 pathways
(i) Glycerol Phosphate pathway (ii)
Monoacylglycerol pathway
(intestinal mucosal cells)
Phosphatidic acid is key
intermediate not only for TAG
synthesis but also other lipids e.g.
phospholipids
 2 molecules of Ketogenesis – Formation of Ketone Bodies
β-Ketothiolase Occurs when acetyl CoA accumulates
Condensation beyond cell’s capacity to oxidise it or to
synthesize fatty acids.
Occurs in the mitochondria, primarily in
the liver.
Acetyl CoA
Extremely important during starvation in
Production of HMG-CoA
which the brain adapts to using ketone
bodies for fuel. Important in Diabetes
mellitus

Lysis Primary ketone body – acetoacetate

Secondary ketone body – acetone & β-
hydroxy butyrate
HMG CoA synthase mitochondrial
Spontaneous fraction – ketogenesis; cytosolic fraction
decarboxylation
Reduction – cholesterol synthesis
HMG CoA lyase present only in liver
Acetoacetate – ketogenic amino acids –
leucine, lysine, phenylalanine, tyrosine.
Utilization of Ketone Bodies (ketolysis)
Ketone bodies are formed in liver but utilized by extrahepatic tissues.

 Heart muscle, renal cortex, skeletal muscle and brain can also utilize the ketone
bodies as alternate sources of energy.
Acetoacetate is activated to acetoacetyl CoA which then enters the beta
 oxidation of fatty acids.
Extrahepatic tissue utilize ketone bodies by converting ß-hydroxybutyrate to
acetoacetate and acetoacetate to acetoacetylcoA.
The first step involves the reversal of the ß-hydroxybutyrate dehydrogenase
reaction, and
the second involves the action of acetoacetate:succinyl CoA transferase
(thiophorase) also called ketoacyl CoA transferase.
Liver cannot convert acetoacetate to acetoacetyl CoA because of the lack of
thiophorase enzyme and hence cannot use ketone bodies for fuel.
Increase in acetyl CoA but low OAA, so acetyl CoA accumulates →
ketogenesis
During starvation, glucagon secreted
Adipose tissue, mobilisation of TAG → fatty acids
Liver, increase breakdown of fatty acids → preferred source of energy
and, at the same time, yields acetyl CoA
Gluconeogenesis stimulated (Pyruvate → OAA →→ Glucose); glucose
conserved for use by tissues dependent on it.

21
Metabolism of Ketone Bodies

Thiophorase
 Ketosis/ Acidosis

When the rate of synthesis of ketone bodies exceeds the ability of

extrahepatic tissues to utilize them, there will be accumulation of
ketone bodies in blood which lowers the pH of blood.
This leads to ketonemia, excretion in urine (ketonuria) and smell of
acetone in breath. All these together consitute the condition known as
ketosis.

Mammals are unable to convert fatty acids into glucose because

there is no pathway for the net production of Oxalaoacetate,
pyruvate, or other gluconeogenic intermediate from acetyl coA.
 LEARNING OUTCOMES

Describe the process of fatty acid synthesis & explain the importance
of citrate in the process.
Describe chain elongation and synthesis of odd-chain fatty acids and
outline desaturation of fatty acids
Outline the biosynthesis of triacylglycerols.
Explain why and how carbohydrates can be used for biosynthesis of
fatty acids but not the other way round for even-chain fatty acids
Discuss why and how ketone bodies are being formed in the liver
Explain why the liver and RBC cannot utilize ketone bodies as source
of energy but other tissues are capable