Introduction

A fatty acid is a carboxylic acid with a long aliphatic chain, which is either
saturated or unsaturated

Fatty acids are the building blocks of the fat in our bodies and in the food we
eat. During digestion, the body breaks down fats into fatty acids, which can
then be absorbed into the blood.

Fatty acid molecules are usually joined together in groups of three, forming a
molecule called a triglyceride.

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 Overview of fatty acid biosynthesis

Occurs in the cytosol of certain animal tissues; e.g., liver and mammary
gland. Also occurs in plants and bacteria.

Uses acetyl-CoA, NADPH as starting materials.

Produces a pool of palmitic acid (16:0) that can be further modified.

Fatty acids play several important roles:

1. Building blocks for phospholipids and glycolipids.

2. Target proteins to membranes.

3. High energy source of fuel.

4. Fatty acid derivatives are used as hormones and intracellular

messengers. 3
 Key players of Fatty Acid Biosynthesis

1. Malonyl-CoA

2. Fatty Acid Synthase

3. Acyl Carrier Protein

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 1. Malonyl-CoA: The intermediate molecule

In fatty acid synthesis, acetyl‐CoA is the direct precursor only of the methyl
end of the growing fatty acid chain. All the other
carbons come from the acetyl group of acetyl‐CoA
but only after it is modified to provide the actual
substrate for fatty acid synthase, malonyl‐CoA.

Malonyl‐CoA contains a 3‐carbon dicarboxylic acid, malonate, bound to

Coenzyme A. Malonate is formed from acetyl‐CoA by the addition of
CO2 using the biotin cofactor of the enzyme acetyl‐CoA carboxylase.

Formation of malonyl‐CoA is the commitment step for fatty acid synthesis,

because malonyl‐CoA has no metabolic role other than serving as a
precursor to fatty acids.
Acetyl-CoA
carboxylase

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 Formation of malonyl-CoA

Acetyl-CoA carboxylase has three activities:

biotin carrier protein

biotin carboxylase
Transcarboxylase

Bicarbonate is phosphorylated, then picked

up by biotin

Biotin swinging arm transfers CO2 to acetyl-

CoA

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 2. Fatty acid synthase (FAS)

The fatty acid synthases (FAS) of

eukaryotes have the component
enzymes linked in a large
polypeptide chain.

Mammalian fatty acid synthase

(FAS) is a large homodimeric
multifunctional enzyme that
regulates the de novo
biosynthesis of long-chain fatty
acids Fatty acid synthase

This cytosolic enzyme catalyzes the formation of 16 carbon (C16) palmitate, from acetyl-
coenzyme A (acetyl-CoA) and malonyl-coenzyme A (malonyl-CoA) in the presence of
NADPH.

The FAS monomer (approximately 270 kDa) contains seven catalytic activities and from
the N-terminus the order is beta-ketoacyl synthase (KS), acetyl/malonyl transacylase
(MAT), beta-hydroxyacyl dehydratase (DH), enoyl reductase (ER), beta-ketoacyl
reductase (KR), acyl carrier protein (ACP), and thioesterase (TE).
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 3. Acyl Carrier Protein

ACP, a single polypeptide chain of 77 residues,

a part of the mammalian Fatty acid synthase,
can be regarded as a giant prosthetic group, a
“macro CoA.”

The intermediates of fatty acid chain are linked

to the sulfhydryl terminus of a
phosphopantetheine group, which is, in turn,
attached to a serine residue of the ACP.

ACP moves from one domain to next during the

fatty acid chain initiation, elongation and
termination events and facilitates the
biosynthesis of fatty acids.

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 Main steps of Fatty Acid Synthesis
1. Initiation Stage

Step 1: loading of acetyl-CoA onto fatty acid synthase

Step 2: loading of malonyl- CoA onto fatty acid synthase

2. Assembly Stage

Step 1: Condensation
Step 2: Reduction
Step 3: Dehydration
Step 4: Reduction

3. Transfer to KS

4. Next cycle begins

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 1. Initiation Stage

Step 1: loading of acetyl-CoA onto fatty

acid synthase

Before the condensation reactions that

build up the fatty acid chain begin, the
two thiol groups on the enzyme complex
must be charged with the correct acyl
groups. The two thiol groups are:

(KS)-Cys–SH
(ACP)-phophopantetheine –SH

Step 1: First, the acetyl group of acetyl-

CoA is transferred to ACP in the reaction
catalysed by the MAT. The acetyl group is
then transferred to the Cys-SH group of
the KS.
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 1. Initiation Stage

Step 2: loading of malonyl- CoA

onto fatty acid synthase

The second reaction, transfer of the

malonyl group from malonyl-CoA to
the –SH group of ACP, is also
catalysed by MAT.

This way charged synthase complex,

the acetyl and malonyl groups are
activated for the chain-lengthening
process.

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 2. Assembly Stage (4 steps)

Step 1: Condensation
Step 2: Reduction
Step 3: Dehydration
Step 4: Reduction of double bond

Step 1: Condensation

Reaction of malonyl group with acetyl group

to form acetoacetyl- ACP

Loss of CO2

Catalysed by β-ketoacyl-ACP synthase (KS)

The acetyl group is transferred from the Cys–

SH group of the enzyme (KS) to the malonyl
group on the –SH of ACP, becoming the
methyl-terminal two-carbon unit of the new
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acetoacetyl group.
 Step 2: Reduction

The acetoacetyl-ACP formed in the condensation step now undergoes

reduction of the carbonyl group at C-3 to form β-hydroxybutyryl-ACP.

The reaction is catalysed by β-ketoacyl-ACP reductase (KR) and the electron

donor is NADPH. 13
 Step 3: Dehydration

The element of water are now removed from C-2 and C-3 of β-hydroxybutyryl-
ACP to Yield a double bond in the product, trans-Δ2-butenoyl-ACP.

The enzyme that catalysed this dehydration is β-hydroxyacyl-ACP-dehydratase

(DH) 14
 Step 4: Reduction of double bond

Finally, the double bond of nis

reduced (saturated) to form
butyryl-ACP by the action of enoyl-
ACP reductase (ER).

NADPH is the electron donor in this

step.

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 3. Transfer to KS

The butryl group is now

transferred from the
phophopantetheine –SH
group of ACP to the Cys-
SH group of KS.

After this the next round

of four reaction cycle
begins.

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 4. Next cycle begins
Another malonyl group is linked to ACP to
initiate next round of four reaction cycle.

The elongation cycle is repeated six more

times, using malonyl–CoA each time, to
produce palmityl–ACP.

The elongation cycles continue until C16-

acyl ACP is formed.

This intermediate is a good substrate for a

thioesterase that hydrolyzes C16-acyl ACP
to yield palmitate and ACP.

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 The Stoichiometry of Fatty Acid Synthesis

The stoichiometry of the synthesis of palmitate is

The equation for the synthesis of the malonyl CoA used in the previous
reaction is

Hence, the overall stoichiometry for the synthesis of palmitate is

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 Citrate Shuttle

The synthesis of palmitate requires

the input of 8 molecules of acetyl
CoA, 14 molecules of NADPH, and 7
molecules of ATP.
Fatty acids are synthesized in the
cytosol, whereas acetyl CoA is
formed from pyruvate in
mitochondria. Hence, acetyl CoA
must be transferred from
mitochondria to the cytosol.
Mitochondria, however, are not
readily permeable to acetyl CoA.
The barrier to acetyl CoA is bypassed by citrate, which carries acetyl groups across the
inner mitochondrial membrane. Citrate is formed in the mitochondrial matrix by the
condensation of acetyl CoA with oxaloacetate. When present at high levels, citrate is
transported to the cytosol, where it is cleaved by ATP-citrate lyase.

Thus, acetyl CoA and oxaloacetate are transferred from mitochondria to the cytosol at the
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expense of the hydrolysis of a molecule of ATP.
 Sources of NADPH for Fatty Acid Synthesis

Oxaloacetate formed in the

transfer of acetyl groups to the
cytosol must now be returned to
the mitochondria. The inner
mitochondrial membrane is
impermeable to oxaloacetate.
Hence, a series of bypass
reactions occurs.

First, the oxaloacetate is

reduced to malate catalysed
by a malate dehydrogenase in the cytosol.

Second, malate is oxidatively decarboxylated by an NADP+-linked malate

enzyme (also called malic enzyme) to form pyruvate.

Cont……

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 Sources of NADPH for Fatty Acid Synthesis

The pyruvate formed in this reaction readily enters mitochondria, where it

is carboxylated to oxaloacetate by pyruvate carboxylase.

Thus, one molecule of NADPH is generated for each molecule of

acetyl CoA that is transferred from mitochondria to the cytosol.

Hence, eight molecules of NADPH are formed when eight molecules of

acetyl CoA are transferred to the cytosol for the synthesis of palmitate.

The additional six molecules of NADPH required for this process come from
the pentose phosphate pathway.

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 Palmitic acid modifications

Palmitate is the starting point for other

fatty acids that use a set of related
reactions to generate the modified chains
and head groups of the lipid classes.

Cell makes a pool of palmitic acid that it

can elongate and/or desaturate in the ER.

Elongation is similar to synthesis of

palmitate because it uses malonyl‐CoA as
an intermediate.

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 Control of fatty acid synthesis

When an organism has more than enough metabolic fuel to meet its energy
needs, the excess is converted to fatty acids and stored as triglycerides.

One of the key regulatory step that control fatty acid synthesis is at the level
of malonyl CoA. The synthesis of malonyl CoA is catalysed by Acetyl-CoA
Carboxylase which is a rate limiting step in the biosynthesis of fatty acids.

Regulation of Acetyl-CoA Carboxylase

The mammalian enzyme is regulated by

A) Allosteric control by local metabolites
B) Phosphorylation
C) Conformational change
1. active conformation is multimeric filamentous structure
2. inactive conformation is non- filamentous form

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 Regulation of Acetyl-CoA Carboxylase
Regulation by Regulation by
Conformational change Allosteric control and
Phosphorylation

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 Nutritional state regulates fatty acid synthesis

During well fed state, the FA synthesis is elevated due to high level of glucose and acetyl-
CoA which is also supported by insulin that activates Acetyl-CoA Carboxylase (ACC) by
dephosphorylation .
In low fed state, the blood glucose is low, that activates PKA which inhibits Acetyl-CoA
Carboxylase by phosphorylation that in turn reduces fatty acid synthesis. 25
Differences between Fatty acid synthesis and degradation (beta oxidation)

Fatty acid (FA) synthesis is not a reversal of the beta oxidation of FA (degradative
pathway). Some important differences between the pathways are:

• Synthesis takes place in the cytosol, however degradation, takes place primarily in
the mitochondrial matrix.

• Intermediates in FA synthesis are covalently linked to the sulfhydryl groups of ACP,

whereas intermediates in FA breakdown are linked to Coenzyme A.

• The enzymes of FA synthesis in higher organisms are joined in a single polypeptide

chain called fatty acid synthase. In contrast, the degradative enzymes do not seem
to be associated.

• The growing FA chain is elongated by the sequential addition of two-carbon

units derived from acetyl CoA. The activated donor of two carbon units in the
elongation step is malonyl ACP.

• The reductant in FA synthesis is NADPH, whereas the oxidants in FA degradation

+
are NAD and FAD.

• Elongation stops at C16 (palmitic acid) 26

 Importance of Fatty acid synthesis
It is a critical anabolic pathway in most organisms. In addition to being the major
component of membranes, fatty acids are important energy storage molecules, and
fatty acyl derivatives possess a variety of physiological functions, including post-
translational modification of numerous proteins. Main functions of fatty acids are:

Energy – high per gram (37 kJ/gram fat)

Transportable form of energy – blood lipids (e.g. triacylglycerol in lipoproteins)

Storage of energy, e.g. in adipose tissue and skeletal muscle

Component of cell membranes (phospholipids)

Insulation - thermal etc

Signals – eicosanoids, gene regulation (transcription)

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Importance of Fatty acids in animals

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 References

Lehninger Principles of biochemistry

Lubert Stryer Biochemistry
Voet and Voet Biochemistry

End

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