Biosynthesis of Fatty Acids

FLOERNCE RAJARETNAM M.Sc., M.Phil.

 Learning Objectives
After reading this chapter you should be able
to:
Describe the pathway of fatty acid synthesis,
& in particular:
the roles of the malonyl-CoA carboxylase
&
the multifunctional enzyme fatty acid
synthase
Outline short-term & long-term regulation of
fatty acid synthesis
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INTRODUCTION
The majority of FAs required by man are supplied
in the diet;
However, pathways for their de novo synthesis
(lipogenesis)
from 2-C compounds are present in many
tissues such as:
liver, brain, kidney, mammary gland, &
adipose tissue

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In general, the pathway of de novo synthesis is
used
in conditions of excess energy, & in
particular, carbohydrate intake.
In such conditions carbohydrates are
converted to fatty acids in the liver
stored as TAG in the adipose tissue

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 Fatty acid synthesis occurs in the cytoplasm of
cells.

Fatty acid synthesis is a stepwise assembly of

acetyl CoA unit (mostly as malonyl CoA) ending with
palmitate (16 C saturated)

The process requires

ATP and
reduced nicotinamide adenine dinucleotide
phosphate (NADPH) as cofactors.

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Principal sources of NADPH + H+
1-Penthose phosphate pathway

2-Reaction by the malic enzyme:

The enzyme is also called NADP dependent
malate dehydrogenase

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Principal
sources of
NADPH +
H+

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Principal sources of Acetyl CoA
1- The degradation of fatty acids
2-The degradation of amino acids.
3-The oxidation of pyruvate.

These three processes occur inside the

mitochondria
the synthesis of fatty acids occurs in the
cytoplasm.
Therefore it is necessary that this acetyl CoA be
transported to the cytoplasm.

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Acetyl CoA is transported from mitochondria to
Cytosol by the following mechanism:

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This process of translocation of citrate from the
mitochondrion to the cytoplasm occurs when the
citrate concentration is high.

Fatty acid synthesis is a stepwise assembly of

acetyl CoA unit (mostly as malonyl CoA) ending
with palmitate (16 C saturated)

During fatty acid synthesis only two enzymes are

necessary:
1-Acetyl CoA carboxylase

2-Fatty Acid synthase

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Step I
Reaction catalyzed by acetyl CoA carboxylase:
This enzyme catalyzes the Carboxylation
reaction of acetyl CoA to malonyl CoA. and
requires
ATP.
enzyme cofactor biotin.

HCO3- + ATP + acetyl-CoA ADP + Pi + malonyl-CoA

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Acetyl-CoA Carboxylase, is the committed (rate
limiting) step of the fatty acid synthesis pathway.

Step II
Reaction catalyzed by Fatty Acid Synthase

A Multienzyme Complex
This multifunctional enzyme catalyzes the seven
different reactions whereby two carbon units from
malonyl-CoA are linked together, ultimately to
form palmitoyl-CoA

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The enzyme complex can exist as both a monomer
and dimer.

The dimeric form is the fully functional form of the

enzyme.

Each of the identical monomer contains

seven catalytic activities and

an acyl carrier protein (ACP) at one end in a

polypeptide chain.

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ACP contains the vitamin pantothenic acid in
the form of 4'-phosphopantetheine (Pant-SH) with
a thiol group at the end.

The other end of the fatty acid synthase

complex carries a thiol group(Cys-SH) from the
amino acid cysteine on the condensing enzyme.

Pant Cyst

SH SH

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The sequence of reactions catalyzed by this
enzyme can be represented by the seven
following steps .

1. A molecule of acetate is transferred from

Acetyl CoA to the SH group of ACP by acetyl
CoA-ACP transacylase (initiation or priming).

2- Next, this 2C fragment is transferred to the

SH-cysteine residue in the active site of the
condensing enzyme.

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3-The now-empty ACP accepts a 3C malonate unit
from malonyl CoA, malonyl CoA-ACP transacylase
catalyzes this reaction

4- Acetyl unit (on the condensing enzyme)

condenses with 2 carbon portion of malonyl unit
on ACP forming acetoacetyl-S- ACP with release
of CO2.
This reaction is catalyzed by -ketoacyl ACP
synthase

Active site on the condensing enzyme is free.

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 acetyl-S-CoA HS-CoA malonyl-S-CoA HS-CoA CO2

Pant Cys Pant Cys Pant Cys Pant Cys

1 2 3
SH SH SH S S S S SH

C O C O C O C O

CH3 CH2 CH3 CH2

1 Malonyl/acetyl-CoA-ACP Transacylase COO C O
2 Malonyl/acetyl-CoA-ACP Transacylase
CH3
3 Condensing Enzyme (-Ketoacyl Synthase)
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5-The -ketone is reduced to an alcohol by NADPH.
6- Dehydration yields a trans double bond.
7- Reduction by NADPH yields a saturated chain.
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8- Following transfer of the growing fatty acid
from Pant to the Condensing Enzyme's cysteine-
SH, the cycle begins again, with another malonyl-
CoA.

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The sequence of reactions, namely condensation,
reduction, dehydration and reduction are repeated
six times, each time incorporating a two-carbon
unit (derived mainly from malonyl CoA) into the
growing fatty acid chain

Note: Acetyl residue successively added is derived

from the 2C atoms of malonyl CoA with the
release of the third C as CO2 EXCEPT the 2
donated by the original acetyl CoA which are
found at the methyl group end of the fatty acid.

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 Product Release
When the fatty acid is 16 carbon atoms long, a
Thioesterase domain catalyzes hydrolysis of the
thioester linking the fatty acid to
phosphopantetheine.

The16-C saturated fatty acid palmitate is the

final product of the Fatty Acid Synthase complex
(but it may produce short chain FAs)

Palmitoyl-ACP + H2O Palmitate + ACP

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 Regulation of FA Synthesis
1. Regulation of Acetyl CoA Carboxylase
Carboxylation of acetyl-CoA to malonyl-CoA is
the committed step of fatty acid
synthesis
This key lipogenic enzyme is subject to both
short term and long term regulation.

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1. Short-term control of fatty acid synthesis is
accomplished by
a) allosteric modifiers of synthetic enzymes.
Two of the most important regulated synthetic
enzymes are the following:
a).Acetyl CoA carboxylase, which is activated by
citrate and inhibited by malonyl CoA and
palmitoyl CoA

b)Fatty acid synthase, which is also inhibited by

palmitoyl CoA.

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Activated by citrate,
which increases in
well-fed state and is
an indicator of a
plentiful supply of
acetyl-CoA

Inhibited by long-
chain acyl-CoA

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b) Covalent modification.
Acetyl CoA carboxylase,
Glucagon promotes its phosphorylation
inactivation
Secretion of insulin promotes its
desphosphorylation activation

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Long-term control of fatty acid synthesis is
accomplished by regulating the synthesis of active
enzymes.
The synthesis of acetyl CoA carboxylase, fatty
acid synthase, citrate lyase, glucose 6 phosphate
dehydrogenase, and the malic enzyme
is decreased during fasting and
increased when glucose is again available in the
diet
Fat-free diets also lead to elevation of the levels
of these enzymes, resulting in increased fatty acid
synthesis

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 -Oxidation & Fatty Acid Synthesis
Compared
-Oxidation Fatty Acid
Pathway Synthesis
Mitochondrial
Pathway location matrix
Cytosol

Acyl carriers Coenzyme -A ACP & Cysteine

(Thiols)

e- Acceptor/Donor FAD & NAD+ NADPH

2-C
Acetyl -CoA Malonyl-CoA (& Acetyl
Product/Donor CoA)

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