ACTIVATION OF Acetyl-CoA

The first step in fatty acid biosynthesis is to

activate acetyl-CoA by the addition of a carbon
dioxide using Acetyl-CoA carboxylase
FORMATION OF MALONYL CoA
The formation of malonyl-CoA from acetyl-CoA irreversible process, committed step catalyzed by
acetyl-CoA carboxylase.

In
animals, the seven reactions of fatty acid synthesis are localized to six
discrete active sites (MAT carries out two reactions, 1a and 1b). The
enzymatic activities are arranged along the polypeptide chain as indicated
in Fig. 20-27a. Several other enzymes exhibit similar multifunctionality, but
none has as many separate catalytic activities as animal fatty acid synthase.
A model of the structure of fatty acid synthase is shown in Fig. 20-27b.
The two subunits form an asymmetric X with two reaction chambers, each
defined by a full set of catalytic domains.
The condensation reaction (2b) requires the juxtaposition of the
sulfhydryl group of an ACP phosphopantetheine and the active site Cys residue, which
are located at opposite ends of the fatty acid synthase
monomer. Experiments with fatty acid synthase mutants indicate that these
groups can interact in the monomer. However, in native fatty acid synthase,
which is a dimer, the interacting groups more often belong to different
monomers. This suggests that the adjacent arms and legs in the structure
shown in Fig. 20-27b belong to different subunits. Presumably, the arms and
legs flex up and down, allowing the long, flexible phosphopantetheine chain
of ACP to transport the substrate between the various catalytic sites. The
flexible lipoyllysyl arms of the pyruvate dehydrogenase multienzyme complex
(Section 17-2B) perform a similar function in that enzyme.Because fatty
acid synthase is a dimer, two fatty acids can be synthesized simultaneously.
In well-nourished individuals, fatty acid synthesis proceeds at a low rate.
However, certain tissues, particularly malignancies, express high levels of

fatty acid synthase and produce fatty acids at a high rate. Consequently,
inhibitors of fatt

There are two major variants of fatty acid synthase:

fatty acid synthase I (FAS I), found in vertebrates and
fungi, and fatty acid synthase II (FAS II), found in plants
and bacteria.
The FAS I found in vertebrates consists of
a single multifunctional polypeptide chain (Mr 240,000).
The mammalian FAS I is the prototype. Seven active
sites for different reactions lie in separate domains (Fig.
213a). The mammalian polypeptide functions as a
homodimer (Mr 480,000). The subunits appear to function
independently.
A somewhat different FAS I is
found in yeast and other fungi, and is made up of two
multifunctional polypeptides that form a complex with
an architecture distinct from the vertebrate systems
With FAS I systems, fatty acid synthesis leads to a
single product, and no intermediates are released.
When the chain length reaches 16 carbons, that product
(palmitate, 16:0; see Table 101) leaves the cycle. Carbons
C-16 and C-15 of the palmitate are derived from
the methyl and carboxyl carbon atoms, respectively, of
an acetyl-CoA used directly to prime the system at the
outset (Fig. 214); the rest of the carbon atoms in the
chain are derived from acetyl-CoA via malonyl-CoA.
FAS II, in plants and bacteria, is a dissociated system;
each step in the synthesis is catalyzed by a separate and
freely diffusible enzyme. Intermediates are also diffusible
and may be diverted into other pathways (such as lipoic
acid synthesis). Unlike FAS I, FAS II generates a variety of
products, including saturated fatty acids of several lengths,
as well as unsaturated, branched, and hydroxy fatty acids.
An FAS II system is also found in vertebrate mitochondria.
The discussion to follow will focus on the mammalian FAS

ATP is used to activate bicarbonate in the form of carboxyphosphate which leads to the
carboxylation of biotin. The activated CO2 group is transferred to acetylCoA to form malonyl
CoA.
Acetyl CoA carboxylase has three domains:
1. A biotin carboxyl group carrier protein.
2. Biotin carboxylase which adds CO2 to biotin.
3. A transcarboxylase which transfers the CO2 group from biotin to acetyl CoA to form
malonyl CoA.
The synthesis of fatty acids, mainly palmitic acid, from acetyl-CoA and
malonyl-CoA involves seven enzymatic reactions. These reactions were
first studied in cell-free extracts of E. coli, in which they are catalyzed by
independent enzymes. Individual enzymes with these activities also occur
in chloroplasts (plant fatty acid synthesis does not occur in the cytosol).
In yeast, fatty acid synthase is a cytosolic, 2500-kD multifunctional enzyme
with the composition _6_6, whereas in animals it is a 534-kD multifunctional
enzyme consisting of two identical polypeptide chains. Presumably,
such proteins evolved by the joining of previously independent genes for
the enzymes.

There are two carriers on this complex.

The first is referred to as ACP1 which acts as a holding station for acetyl or

fatty acyl- groups. In either case they are bound to a cysteinyl

sulphhydryl group.
The second, ACP2, binds the growing fatty acyl chain during the
condensation and reduction reactions of the cycle. In this case the acyl
group is carried on a long phosphopantetheine prosthetic group.

Acyl carrier protein (ACP) is the shuttle that

holds the system together. The Escherichia coli ACP is
a small protein (Mr 8,860) containing the prosthetic
group 49-phosphopantetheine (Fig. 215; compare
this with the panthothenic acid and _-mercaptoethylamine
moiety of coenzyme A in Fig. 838). The 4_-phosphopantetheine
prosthetic group of E. coli ACP is
believed to serve as a flexible arm, tethering the growing
fatty acyl chain to the surface of the fatty acid synthase
complex while carrying the reaction intermediates from
one enzyme active site to the next. The ACP of mammals
has a similar function and the same prosthetic group; as
we have seen, however, it is embedded as a domain in a
much larger multifunctional polypeptide.

Fatty Acid Synthase Receives the Acetyl

and Malonyl Groups
Before the condensation reactions that build up the fatty
acid chain can begin, the two thiol groups on the enzyme
complex must be charged with the correct acyl groups
(Fig. 216, top). First, the acetyl group of acetyl-CoA istransferred to ACP in a reaction
catalyzed by the malonyl/
acetyl-CoAACP transferase (MAT in Fig. 216)
domain of the multifunctional polypeptide. The acetyl
group is then transferred to the Cys SH group of the
_-ketoacyl-ACP synthase (KS). The second reaction,
transfer of the malonyl group from malonyl-CoA to the
SH group of ACP, is also catalyzed by malonyl/acetylCoAACP transferase. In the charged synthase complex,
the acetyl and malonyl groups are activated for the
chain-lengthening process. The first four steps of this
process are now considered in some detail; all step
numbers refer to Figure 216.

These are priming reactions in which the synthase is loaded with

the precursors for the condensation reaction. In mammals,
malonyl/acetyl-CoA-ACP transacylase (MAT) catalyzes two similar
reactions at a single active site: An acetyl group originally
linked as a thioester in acetyl-CoA is transferred to ACP (1a), and
a malonyl group is transferred from malonyl-CoA to ACP (1b).
2. The _-ketoacyl-ACP synthase (KS; also known as condensing enzyme)

first transfers the acetyl group from ACP to an enzyme Cys

residue (2a). In the condensation reaction (2b), the malonyl-ACP
is decarboxylated, and the resulting carbanion attacks the acetylthioester
to form a four-carbon acetoacetyl-ACP. The decarboxylation
reaction drives the condensation reaction

The first reaction, catalyzed by Acetyl-CoA:ACP transacylase, transfers an acetyl

group from Coenzyme A to the cysteinyl-S on ACPl.
Next, a malonyl-group is transferred from a Coenzyme A to the
pantetheinyl-S of ACP2 by Malonyl-CoA:ACP transacylase
Now the carbon dioxide leaves the malonyl group, with the electrons
from its bond attacking the acyl group on ACP1. This reaction is
catalyzed by Ketoacyl-ACP synthase.
Now have a beta-ketoacyl group ready to go through the reverse of the reactions
of beta-oxidation.
Thus, the keto-group is reduced to an alcohol using NADPH (betaketoacylACP reductase),
followed by the elimination of the alcohol (Enoyl-ACP hydrase) to give
the cis-2,3-enoyl group.
The enoyl is then reduced with NADPH substituting for FADH2 (EnoylACP
reductase) to give the saturated acyl group.
Finally the acyl group is transferred from the pantotheinyl-S of ACP2 to
the cysteinyl-S on ACPl (ACP-acyltransferase) leaving ACP2 available to
pick up the next malonyl moiety.
After seven turns of the cycle palmitate is released.
Elongation by the fatty acid synthase complex stops upon formation of
palmitate (16 C). Further elongation and the formation of double bonds
are carried out by other enzyme systems.
Summary of Fatty Acid Biosynthesis
Acetyl CoA + 7 malonyl CoA + 14 NADPH + 20 H+
Palmitate + 7 C02 + 14 NADP+ + 8 CoASH + 6
H20
The equation for the synthesis of the malonyl CoA used in the abave reaction is
7 Acetyl CoA + 7 C02 + 7 ATP 7 malonyl CoA + 7 ADP + 7 Pi + 14 H+
The overall reaction for the synthesis of palmitate is
8 Acetyl CoA + 7 ATP + 14 NADPH + 6H+
Palmitate + 14 NADP + 8 CoASH + 6 H20 + 7 ADP + 7
Pi