Oxidation of Fatty Acids

Fatty Acid Oxidation is the primary source of energy for many tissues with the exception of the
central nervous system and circulating RBC.
Process by which fatty acids are broken down to obtain energy:
• The fatty acid must be activated
• The fatty acid must be transported into the mitochondrial matrix by a shuttle mechanism
• The fatty acid must be repeatedly oxidized, cycling thru a series of 4 reactions, to produce
acetyl CoA, FADH2 and NADH

Fatty Acid Activation

Site: Outer Mitochondrial Membrane
Reactants: Fatty acid, CoA and ATP
Enzyme: Acyl CoA Synthase
Fatty acid is converted to Acyl CoA
Activated Fatty Acid CoA: Acyl CoA
This reaction requires the expenditure of two high-energy phosphate bonds from a single ATP
molecule; the ATP is converted to AMP rather than ADP, and the resulting pyrophosphate (PPi)
is hydrolyzed to 2Pi.

Fatty Acid Transport

Acyl CoA is too large to pass through the inner mitochondrial membrane to the mitochondrial
matrix, where the enzymes needed for fatty acid oxidation are located. A shuttle mechanism
involving the molecule carnitine allows the entry of acyl CoA into the matrix. The acyl group is
transferred to a carnitine molecule, which carries it through the membrane. The acyl group is then
transferred from the carnitine back to a CoA molecule.
Regulation of Carnitine Shuttle
● This carnitine shuttle is a rate limiting step in the oxidation of fatty acids in the
mitochondria and thus fatty acid oxidation can be regulated at this step.
● Malonyl CoA, an intermediate of fatty acid synthesis present in the cytosol is an inhibitor
of carnitine acyltransferase I. This indicates that when fatty acid synthesis is in progress,
oxidation of fatty acid cannot occur at the same time as the carnitine shuttle is impaired
by inhibition of carnitine acyltransferase I.

Beta-Oxidation Pathway
A repetitive series of four biochemical reactions degrading Acyl CoA to Acetyl CoA by removing
two carbon atoms at a time, with FADH2 and NADH also being produced. Each repetition of the
four-reaction sequence generates an Acetyl CoA molecule and an Acyl CoA molecule that has two
fewer carbon atoms.

For a saturated fatty acid, the b-oxidation pathway involves the following functional group changes
at the b carbon and the following reaction types.
  Beta Oxidation of Saturated Fatty Acids
Step 1: Oxidation (dehydrogenation)
Hydrogen atoms are removed from the a and b carbons, creating a double bond between these two
carbon atoms. FAD is the oxidizing agent, and a FADH2 molecule is a product. Enzyme: Acyl
CoA Dehydrogenase (Stereospecific)

Step 2: Hydration
A molecule of water is added across the trans double bond, producing a secondary alcohol at the
b-carbon position. Again, the enzyme involved is stereospecific in that only the L-hydroxy isomer
is produced from the trans double bond.
Enzyme: Enoyl CoA Hydratase

Step 3: Oxidation (dehydrogenation)

The beta-hydroxy group is oxidized to a ketone functional group with NAD+ serving as the
oxidizing agent. The required enzyme exhibits absolute stereospecificity for the L-isomer.
Enzyme: B-hydroxyacyl CoA Dehydrogenase
Step 4: Chain Cleavage
The fatty acid chain is broken between the a and b carbons by reaction with a coenzyme A
molecule. The result is an acetyl CoA molecule and a new acyl CoA molecule that is shorter by
two carbon atoms than its predecessor.
Enzyme: Thiolase

The new acyl CoA molecule (now shorter by two carbons) is recycled through the same set of four
reactions again. This yields another acetyl CoA, a two-carbon-shorter new acyl CoA, FADH2, and
NADH. Recycling occurs again and again, until the entire fatty acid is converted to acetyl CoA.
Thus the fatty acid carbon chain is sequentially degraded, two carbons at a time.

The number of acetyl CoA molecules produced in the b-oxidation pathway is equal to half the
number of carbon atoms in the fatty acid.
Acetyl CoA Molecules = no. of C-atoms FA/2
The number of repetitions of the b-oxidation pathway that are needed to produce the acetyl CoA
is always one less than the number of acetyl CoA molecules produced
B-Oxidation cycle = no. of Acetyl CoA - 1
The number of FADH2 and NADH molecules produced are equal to the number of repetitions of
the Beta-oxidation cycle
B-Oxidation cycle = no. of FADH2 and NADH
ATP YIELD IN SATURATED FATTY ACIDS
9 repetitions of the B-oxidation pathway are required for the oxidation of Arachidonic (eicosanoic
acid) or C20 fatty acid.
These 9 repetitions of the pathway produce 10 acetyl CoA molecules, 9 FADH2 molecules, and 9
NADH molecules. It is processed further in common metabolic pathway (citric acid cycle, electron
transport chain, and oxidative phosphorylation) leads to ATP production as follows:
10 Acetyl CoA x 10 ATP/Acetyl CoA = 100 ATP
9 NADH x 3ATP/NADH = 27 ATP
9 FADH2 x 2ATP/FADH2 = 18 ATP
TOTAL ATP YIELD: 145 ATP

 Beta Oxidation of Unsaturated Fatty Acids

β-oxidation pathway for unsaturated fatty acids includes two additional enzymes isomerase and
reductase. It still follows the four repetitive steps much like how saturated fatty acids are oxidized.
Just like the saturated fatty acids cross the mitochondrial membrane with the help of carnitine
shuttle. Unsaturated fatty acids also reach the mitochondrial matrix as fatty acyl-CoA.
β-oxidation of oleic acid (MUFA) by the enzymes present in the mitochondrial matrix is as shown:
β-oxidation of linoleic acid (PUFA) by the enzymes present in the mitochondrial matrix is as
shown:
Unsaturated Fatty Acid Oxidation
THE SAME IS TRUE WITH UNSATURATED FATTY ACID OXIDATION:

Acetyl CoA Molecules = no. of C-atoms FA/2

B-Oxidation cycle = no. of Acetyl CoA - 1
B-Oxidation cycle = no. of FADH2 and NADH

ATP YIELD IN UNSATURATED FATTY ACIDS (OLEIC ACID)

8 repetitions of the B-oxidation pathway are required for the oxidation of Oleic Acid or C18 fatty
acid.
These 8 repetitions of the pathway produce 9 acetyl CoA molecules, 8 FADH2 molecules, and 8
NADH molecules. It is processed further in common metabolic pathway (citric acid cycle, electron
transport chain, and oxidative phosphorylation) leads to ATP production as follows:
9 Acetyl CoA x 10 ATP/Acetyl CoA = 90 ATP
8 NADH x 3ATP/NADH = 24 ATP
8 FADH2 x 2ATP/FADH2 = 16 ATP
TOTAL ATP YIELD: 130 ATP

ATP YIELD IN UNSATURATED FATTY ACIDS (LINOLEIC ACID)

8 repetitions of the B-oxidation pathway are required for the oxidation of Linoleic Acid or C18
fatty acid.
These 8 repetitions of the pathway produce 9 acetyl CoA molecules, 8 FADH2 molecules, and 8
NADH molecules. It is processed further in common metabolic pathway (citric acid cycle, electron
transport chain, and oxidative phosphorylation) leads to ATP production as follows:
9 Acetyl CoA x 10 ATP/Acetyl CoA = 90 ATP
8 NADH x 3ATP/NADH = 24 ATP
8 FADH2 x 2ATP/FADH2 = 16 ATP
1NADP+ = 4 ATP
TOTAL ATP YIELD: 134 ATP
Beta Oxidation Overall Reaction
Cn-acyl-CoA + FAD + (NAD+) + H2O + CoA-SH
→
Cn-acyl-CoA + FADH2 + NADH + (H+) + acetyl-CoA

 Beta Oxidation of Odd Chain Fatty Acids

Just the way even chain carbon length fatty acids undergo B-Oxidation, odd carbon chain fatty
acids undergo the same pathway leaving propionyl-CoA as an end product along with acetyl-
CoA molecules. This propionyl-CoA is formed because during the last B-oxidation cycle the
fatty acyl-CoA with five carbons is cleaved to give a three carbon compound and a two carbon
compound unlike even carbon chain length fatty acid. Acetyl CoA enters the Kreb’s cycle but
propionyl-CoA has a different fate.

Step 1: Carboxylation
The propionyl CoA is converted into D-methylmalonyl CoA by the activity of the enzyme
propionyl CoA carboxylase. This enzyme depends on a water soluble B- vitamin called Biotin.
Step 2: Epimerization
The D conformation is enzymatically converted into the L conformation by methylmalonyl-
CoA epimerase.

Step 3: Isomerization
The enzyme called methylmalonyl CoA mutase catalyzes an intramolecular rearrangement
reaction in which the –CO-S-CoA group migrates from C2 onto the methyl group in exchange
for a hydrogen. This arrangement occurs only in the presence of vitamin B12.

The Succinyl-CoA formed can then enter the Kreb’s cycle.

 The number of B-Oxidation cycle, NADH, FADH2 and the number of Acetyl CoA molecules
can be found using:
(No. of C-atoms FA/2) – 1.5

ATP YIELD IN ODD CHAIN FATTY ACIDS

7 repetitions of the B-oxidation pathway are required for the oxidation of C17 fatty acid.
These 7 repetitions of the pathway produce 7 acetyl CoA molecules, 7 FADH2 molecules,
and 7 NADH molecules. It is processed further in common metabolic pathway (citric acid
cycle, electron transport chain, and oxidative phosphorylation) leads to ATP production as
follows:
7 Acetyl CoA x 10 ATP/Acetyl CoA = 70 ATP
7NADH x 3ATP/NADH = 21 ATP
7 FADH2 x 2ATP/FADH2 = 14 ATP

ATP Calculation for Propionyl

1 NADH x 2.5 ATP/NADH = 2.5 ATP
1 FADH x 1.5ATP/FADH = 1.5 ATP
1 GTP = 1 ATP
1 Propionyl = 5 ATP
Even part of the Chain = 105 ATP
ATP yield for Odd Chain = 110 ATP