Chapter 22.

Fatty Acid Metabolism

BMB 312: Lipid Metabolism

Target proteins to membranes

High energy source of fuel

Fatty acid derivatives are used as hormones and

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 Introduction
Overview of
fatty acid
synthesis

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 1. Triglycerides
Triglycerides are a highly concentrated store
of energy
9 kcal/g vs 4 kcal/g for glycogen
Glycogen is also highly hydrated, 2 g H2O/g
glycogen

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 1.1 Pancreatic Lipases
Dietary triacylglycerols must be broken down
before being absorbed by the intestines.
Bile salts, which act as detergents, are used to
solublize the triacylglycerols

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 1.1 Pancreatic Lipases
Dietary triacylglycerols must be broken down
before being absorbed by the intestines.
Bile salts, which act as detergents, are used to
solublize the triacylglycerols

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 1.1 Pancreatic Lipases
Pancreatic lipases hydrolyze the ester bonds
of the triacylglycerols while in the
micelles.

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 2. Utilization of Fatty Acids as Fuel
Three stages of processing
Triglycerols are degraded to fatty acids and
glycerol in the adipose tissue and transported to
other tissues.
Fatty acids are activated and transported into the
mitochondria.
Fatty acids are broken down into two-carbon
acetyl–CoA units and fed into the citric acid cycle.

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 2.1 Breakdown of Triacylglycerols
In the adipose tissue, lipases are activated
by hormone signaled phosphorylation

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 2.1 Breakdown of Triacylglycerols
The lipases break the
triacylglycerols down to
fatty acids and glycerol
The fatty acids are
transportred in the blood by
serum albumin

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 2.1 Breakdown of Triacylglycerols
The glycerol is absorbed by the liver and
converted to glycolytic intermediates.

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Fatty acid oxidation is called “β-oxidation”
because the bond between the α (C2) and β
carbon (C3) of the fatty acid is broken
during each round of the cycle,
 2.2 Activation of Fatty Acids
Acyl CoA synthetase reaction occurs in the on
the mitochondrial membrane.

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2.3 Transport into Mitochondrial Matrix
Carnitine carries
long-chain
activated fatty
acids into the
mitochondrial
matrix

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2.3 Transport into Mitochondrial Matrix
Carnitine carries long-chain activated fatty
acids into the mitochondrial matrix

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 2.4 Fatty acid oxidation
Each round in fatty acid
degradation involves
four reactions
1. oxidation to
trans-∆2-Enoly-CoA

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 2.4 Fatty acid oxidation
Each round in fatty acid
degradation involves
four reactions
2. Hydration to L–3–
Hydroxylacyl CoA

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 2.4 Fatty acid oxidation
Each round in fatty acid
degradation involves
four reactions
3. Oxidation to
3–Ketoacyl CoA

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 2.4 Fatty acid oxidation
Each round in fatty acid
degradation involves
four reactions
4. Thiolysis to produce
Acetyl–CoA

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 2.4 Fatty acid oxidation
Each round in fatty acid
degradation involves
four reactions
The process repeats itself

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 2.4 Fatty acid oxidation
Each round in fatty acid degradation involves
four reactions

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 2.5 ATP Yield
The complete oxidation of the sixteen
carbon palmitoyl–CoA produces 106 ATP's

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 3.1 Special Cases
Unsaturated fatty acids
(monounsaturated)

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 3.1 Special Cases
Unsaturated fatty acids (polyunsaturated)

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 3.2 Odd-Chain

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Fatty Acid Biosynthesis
• Synthesis takes place in the cytosol
• Intermediates covalently linked to acyl
carrier protein
• Activation of each acetyl CoA.
• acetyl CoA + CO2 Malonyl CoA
• Four-step repeating cycle,
extension by 2-carbons / cycle – Condensation
– Reduction – Dehydration -Reduction
Fatty acid synthesis
• The enzymes of fatty acid synthesis are packaged together
in a complex called as fatty acid synthase (FAS).
• The product of FAS action is palmitic acid. (16:0).
• Modifications of this primary FA leads to other longer (and
shorter) FA and unsaturated FA.
• The fatty acid molecule is synthesized 2 carbons at a time
• FA synthesis begins from the methyl end and proceeds
toward the carboxylic acid end.
Thus, C16 and C15 are added first and C2 and C1 are added
last.
• C15 and C16 are derived directly from acetylCoA. For
further step-wise 2-carbon extensions, acetylCoA is first
activated to malonyl CoA, a 3-carbon compound, by the
addition of a CO2.
Citrate Shuttle
• FAs are synthesized in the cytoplasm from acetylCoA
• AcetylCoA generated from pyruvate by the action of PDH
and by β-oxidation of fatty acids is in the mitochondria.
• For fatty acid biosynthesis, acetylCoA has to be transported
from the mitochondria to the cytoplasm. This is done via a
shuttle system called the Citrate Shuttle.
• AcetylCoA reacts with oxaloacetate to give citrate. A
tricarboxylate translocase transports citrate from
mitochondria to cytosol.
• In the cytosol, citrate is cleaved back to oxaloacetate and
acetylCoA. This reaction is catalyzed by ATP-citrate lyase
and requires the hydrolysis of one molecule of ATP.
Reaction catalyzed by Acetyl CoA Carboxylase
MalonylCoA
• Malonyl CoA is synthesized by the action of acetylCoA
carboxylase. Biotin is a required cofactor.
• CH 3COSCoA + CO2 + ATP OOC-CH2-COSCoA + ADP +Pi
(enzyme: acetylCoA carboxylase)
• This is an irreversible reaction. AcetylCoA carboxylation is
a rate-limiting step of FA biosynthesis.
• AcetylCoA carboxylase is under allosteric regulation.
Citrate is a positive effector and palmitoyl CoA is a negative
effector
Fatty Acid Synthase (FAS)
• FAS is a polypeptide chain with multiple domains, each with
distinct enzyme activities required for fatty acid
biosynthesis.
• ACP: Recall that CoA is used as an activator for β oxidation.
For fatty acid biosynthesis, the activator is a protein called
the acyl carrier protein (ACP). It is part of the FAS
complex. The acyl groups get anchored to the CoA group of
ACP by a thioester linkage
• Condensing enzyme/β-ketoacyl synthase (K-SH). Also part
of FAS, CE has a cysteine SH that participates in thioester
linkage with the carboxylate group of the fatty acid.
• During FA biosynthesis, the growing FA chain alternates
between K-SH and ACP-SH
Step-wise reactions
1.The acetyl group gets transferred from CoA to ACP by acetyl
CoA-ACP transacylase.
2.The acetyl (acyl) group next gets transferred to the K arm
of FAS complex.
3.Next, the malonyl group gets transferred from CoA to ACP
by malonyl CoA ACP transacylase. This results in both arms
of FAS occupied forming acylmalonyl-ACP.
4.The COO group of malonyl ACP is removed as CO2, the
acetyl group (C16 and C15) gets transferred to the alpha
carbon of malonyl ACP. This results in 3keto acyl ACP.
 3.5 Ketone Bodies
Use of fatty acids in the citric acid cycle
requires carbohydrates for the the
production of oxaloacetate.
During starvation or diabetes, OAA is used to
make glucose
Fatty acids are then used to make ketone bodies
(acetoacetate and D–3–hydroxybutarate)

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 3.5 Ketone Bodies
Ketone bodies, acetoacetate and 3–
hydroxybutarate are formed from Acetyl–
CoA

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 3.6 Ketone Bodies as a Fuel Source
The liver is the major source of ketone
bodies.
It is transported in the blood to other tissues

Acetoacetate in the tissues

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 3.7 Fatty Acids Cannot be Used to
Synthesize Glucose
Even though the citric acid cycle
intermediate oxaloacetate can be used to
synthesize glucose, Acetyl–CoA cannot be
used to synthesize oxaloacetate.
The two carbons that enter the citric acid cycle as
Acetyl–CoA leave as CO2.

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 4. Fatty Acid Synthesis.
Fatty acid are synthesized and degraded by
different pathways.
Synthesis takes place in the cytosol.
Intermediates are attached to the acyl carrier
protein (ACP).
In higher organisms, the active sites for the
synthesis reactions are all on the same polypeptide.
The activated donor in the synthesis is malonyl–ACP.
Fatty acid reduction uses NADPH + H+.
Elongation stops at C16 (palmitic acid)

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 4.1 Formation of Malonyl Coenzyme A
Formation of malonyl–CoA is the committed
step in fatty acid synthesis.

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 4.2 Acyl Carrier Protein
The intermediates in fatty acid synthesis are
covalently linked to the acyl carrier protein
(ACP)

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 4.3 Elongation
In bacteria the enzymes that are involved in
elongation are separate proteins; in higher
organisms the activities all reside on the
same polypeptide.
To start an elongation cycle, Acetyl–CoA and
Malonyl–CoA are each transferred to an acyl carrier
protein

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 4.3 Elongation
Acyl-malonyl ACP
condensing enzyme forms
Acetoacetyl-ACP.

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 4.3 Elongation
The next three reactions are similar to the
reverse of fatty acid degradation, except
The NADPH is used instead of NADH and FADH2
The D–enantiomer of Hydroxybutarate is formed instead of the L–enantiomer

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 4.3 Elongation
The elongation cycle is repeated six more
times, using malonyl–CoA each time, to
produce palmityl–ACP.

A thioesterase then cleaves the palmityl–CoA

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4.4 Multifunctional Fatty Acid Synthase
Domain 1
Substrate entry (AT & MT) and condensation unit
(CE)
Domain 2
Reduction unit (DH, ER & KR)
Domain 3
Palmitate release unit (TE)

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4.4 Multifunctional Fatty Acid Synthase

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 4.4 Multifunctional Fatty Acid
Synthase
46 A NEW STRUCTURE
 4.5 Fatty Acid Synthase Mechanism

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 4.6 Stoichiometry of FA synthesis
The stoichiometry of palmitate synthesis:
Synythesis of palmitate from Malonyl–CoA

Synthesis of Malonyl–CoA from Acetyl–CoA

Overall synthesis

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 4.7 Citrate Shuttle
Acetyl–CoA is synthesized in the
mitochondrial matrix, whereas fatty acids
are synthesized in the cytosol
Acetyl–CoA units are shuttled out of the mitochondrial matrix as citrate:

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 4.8 Sources of NADPH
+
The malate dehydrogenase and NADP –linked
malate enzyme reactions of the citrate
shuttle exchange NADH for NADPH

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4.9 Fatty Acid Synthase Inhibitors (skip)

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 4.10 Variations on a Theme (skip)

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 5. Regulation of Fatty Acid Synthesis
Regulation of Acetyl carboxylase
Global
+ insulin
- glucagon
- epinephrine
Local
+ Citrate
- Palmitoyl–CoA
- AMP

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 5.1 Regulation of Fatty Acid Synthesis

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 6. Elongation and Unsaturation
Endoplasmic reticulum systems introduce
double bonds into long chain acyl–CoA's
Reaction combines both NADH and the acyl–CoA's to
reduce O2 to H2O.

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 6.1 Elongation and Unsaturation
Elongation and unsaturation convert
palmitoyl–CoA to other fatty acids.
Reactions occur on the cytosolic face of the
endoplasmic reticulum.
Malonyl–CoA is the donor in elongation reactions

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 6.2 Eicosanoid Hormones
Eicosanoid horomones are synthesized from
arachadonic acid (20:4).
Prostaglandins
20-carbon fatty acid containing 5-carbon ring
Prostacyclins
Thromboxanes
Leukotrienes
contain three conjugated double bonds

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 6.2 Eicosanoid Hormones

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 6.2 Eicosanoid Hormones

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 6.2 Eicosanoid Hormones

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