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Beta Oxidation of Fatty acids

Overview of fatty acid and triglyceride

Fatty acids can be define as a carboxylic acid with an aliphatic hydrocarbon chain that is
either saturated or unsaturated. They are the building blocks of the fat in our bodies and in the
food we eat. 3 molecules of fatty acids reacts with glycerol to form fat (triglycerides) in
esterification reaction. During digestion, the body breaks down fats into fatty acids, which can
then be absorbed into the blood. Fatty acids can be classified based on length of hydrocarbon,
presence of alkene group, based on even and odd chain, essential and non-essential amino acid

Triacylglycerols (fats) are the most abundant source of energy and provide energy twice as much
as carbohydrates and proteins. This is achieved because the fatty acids which are present in the
triacylglycerols are already in the reduced form. To convert fats into energy, the fatty acids of the
digested fats or the stored fats (have to be first activated and transported to the mitochondrial
matrix as all the enzymes required for metabolism (oxidation) are present there

Fatty acid oxidation is the mitochondrial aerobic process of breaking down a fatty acid into
acetyl-CoA units. Fatty acids released in the digestion of triglycerides and other lipids are
broken down in a series of sequential reactions accompanied by the gradual release of usable
energy. Some of these reactions are oxidative and require nicotinamide adenine dinucleotide
(NAD+) and flavin adenine dinucleotide (FAD). The enzymes that participate in fatty acid
catabolism are located in the mitochondria, along with the enzymes of the citric acid cycle, the
electron transport chain, and oxidative phosphorylation. This localization of enzymes in the
mitochondria is of the utmost importance because it facilitates efficient utilization of energy
stored in fatty acids and other molecules.

Activation and transportation of fatty acid to mitochondrial matrix

Fatty acid oxidation is initiated on the outer mitochondrial membrane. To be oxidized, they must
be transported to the mitochondria matrix by attaching a coenzyme A to it. Therefore the fatty
acids must first be activated by conversion to an energy-rich fatty acid derivative of coenzyme A
called fatty acyl-coenzyme A (CoA). The activation is catalyzed by acyl-CoA synthetase. For
each molecule of fatty acid activated, one molecule of coenzyme A and one molecule of
adenosine triphosphate (ATP) are used, equaling a net utilization of the two high-energy bonds
in one ATP molecule (which is therefore converted to adenosine monophosphate [AMP] rather
than adenosine diphosphate [ADP]):
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The fatty acyl-CoA combines with a carrier molecule called carnitine in the cytosol to form
acyl-carnitine diffuses to the inner mitochondrial membrane in a reaction catalyzed by carnitine
acyltransferase 1 and the translocase in the inner mitochondrial membrane transported it to
mitochondrial matrix where it is converted back to the fatty acyl-CoA by carnitine acyltransferase
2.

Role of carnitine in the transport of Acyl-CoA from cytosol to the mitochondrial matrix.
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Steps in the β-Oxidation of Fatty Acids

Further oxidation of the fatty acyl-CoA occurs in the mitochondrial matrix via a sequence of four
reactions known collectively as β-oxidation because the β-carbon undergoes successive
oxidations in the progressive removal of two carbon atoms from the carboxyl end of the fatty
acyl-CoA. The process involves dehydrogenation, hydration, oxidation and thiolysis.

The first step in the catabolism of fatty acids is the formation of an alkene in an oxidation
reaction catalyzed by acyl-CoA dehydrogenase. In this reaction, the coenzyme FAD accepts two
hydrogen atoms from the acyl-CoA, one from the α-carbon and one from the β-carbon, forming
reduced flavin adenine dinucleotide (FADH 2) that supplies energy to form 1.5–2 molecules of
ATP.

Next, the alkene is hydrated to form a secondary alcohol in a reaction catalyzed by enoyl-CoA
hydratase.

The secondary alcohol is then oxidized to a ketone by β-hydroxyacyl-CoA dehydrogenase, with

NAD+ acting as the oxidizing agent form NADH . The reoxidation of each molecule of NADH to
NAD+ by the electron transport chain furnishes 2.5–3 molecules of ATP.

The final reaction is cleavage of the β-ketoacyl-CoA by a molecule of coenzyme A. The products
are acetyl-CoA and a fatty acyl-CoA that has been shortened by two carbon atoms. The reaction
is catalyzed by thiolase.
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Example of beta oxidation of saturated fatty acid (palmitic acid)

ATP GENERATION FOR PALMITIC ACID

2 ATP used= -2 ATP ( Two ATP used to activate fatty acid)
7 FADH= 7×2 ATP=14
7 NADH= 7×3 ATP=21
8 acetyl coA = 8×12 ATP= 96
96+14+21-2=129
Palmitic acid will undergo 7 cycles

Beta oxidation of monounsaturated and polyunsaturated fatty acid

Monounsaturated fatty acid consists of a double bond. Beta oxidation of monounsaturated fatty
acid involves most of the reactions same as found in beta oxidation of saturated fatty acid.
However, an extra enzyme enoyl coA isomerase is needed. similarly, due to presence of two or
more double bonds, beta oxidation of poly unsaturated fatty acid requires two additional
enzymes- enoyl coA isomerase and 2, 4- dienoyl coA reductase.

See the hand written paper

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Beta oxidation of polyunsaturated fatty acid

Calculation of Energy of
7 NADH= 7×2=14 ATP
7 FADH = 7×2= 14 ATP
9 acetyl COA = 9×12=108
Total =143 ATP
2 ATP used for fatty acid activation= -2
Net total= 143-2=141 ATP
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Beta oxidation of Odd Chain fatty acid (OCFA)

Odd-chain fatty acids are those fatty acids that contain an odd number of carbon atoms. In additn
to being classified according to their saturation or unsaturation, fatty acids are also classified
according to their odd or even numbers of constituent carbon atoms. OCFAs are found
particularly in ruminant fat and milk (e.g. heptadecanoic acid) and some plants. Example are
pentadecanoic acid (15:0)( pentadecylic acid) and heptadecanoic acid (17:0)( Margaric acid)
Odd-chain fatty acids undergo beta-oxidation in the same manner as even chain fatty acids;
however, once a five-carbon chain remains, the final spiral of beta-oxidation will yield one
molecule of acetyl CoA and one molecule of propionyl CoA. The propionyl CoA is ultimately
transformed via a three-step process into succinyl CoA, which can then be incorporated into the
citric acid cycle.
These three steps process involves three different enzymes - propionyl CoA carboxylase,
methylmalonyl CoA racemase and methylmanlonyl CoA mutase.
Steps of beta oxidation of odd fatty acid
 Carboxylation of propionyl COA to make D-methylmalonyl-CoA by propionyl COA
carboxylase
 Isomerization of D-methylmalonyl to L-methylmalonyl-CoA by methylmalonyl racemase
 Rearrangement of L-methylmalonyl-coA to form succinyl-CoA. Ths last step of the
process utilizes the enzyme methylmalonyl-CoA mutase, which uses the B12 coenzyme
in its catalytic cycle. Succinyl-CoA can then be metabolized in the citric acid cycle.

Assignment
Succinctly write on short chain fatty acids and their biochemical relevance?
KETONE BODIES

Ketogenesis is the biochemical process through which organisms produce ketone

bodies by breaking down fatty acids and ketogenic amino acids.
Ketone bodies are produced by the liver and used peripherally as an energy source when glucose
is not readily available. Ketone bodies are water-soluble molecules or compounds that contain
the ketone groups produced from fatty acids by the liver. Ketone bodies are produced by the liver
during periods of caloric restriction of various scenarios: low food intake (fasting), carbohydrate
restrictive diets, starvation, prolonged intense exercise, alcoholism, or during untreated (or
inadequately treated) type 1 diabetes mellitus. Ketone bodies are produced in liver cells by the
breakdown of fatty acids. They are released into the blood after glycogen stores in the liver have
been depleted. (Glycogen stores typically are depleted within the first 24 hours of fasting.

The two main ketone bodies are acetoacetate and 3-beta-hydroxybutyrate (3HB), while acetone
is the third, and least abundant, ketone body. Ketones are always present in the blood and their
levels increase during fasting and prolonged exercise.
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In the liver, most of the acetyl-CoA obtained from fatty acid oxidation is oxidized by the citric
acid cycle. However, under certain metabolic conditions such as starvation or diabetes
mellitus, the rate of fatty acid oxidation increases to provide energy. This leads to an increase in
the concentration of acetyl-CoA. The increased acetyl-CoA cannot be oxidized by the citric acid
cycle because of a decrease in the concentration of oxaloacetate, which is diverted to glucose
synthesis. Therefore, some of the acetyl-CoA is used to synthesize a group of compounds
known as ketone bodies: acetoacetate, β-hydroxybutyrate, and acetone. Two acetyl-CoA
molecules combine, in a reversal of the final step of β-oxidation, to produce acetoacetyl-CoA.
The acetoacetyl-CoA reacts with another molecule of acetyl-CoA and water to form β-hydroxy-
β-methylglutaryl-CoA, which is then cleaved to acetoacetate and acetyl-CoA. Most of the
acetoacetate is reduced to β-hydroxybutyrate, while a small amount is decarboxylated to carbon
dioxide and acetone.

The acetoacetate and β-hydroxybutyrate synthesized by the liver are released into the blood for
use as a metabolic fuel (to be converted back to acetyl-CoA) by other tissues, particularly the
kidney and the heart. Thus, during prolonged starvation, ketone bodies provide about 70% of the
energy requirements of the brain. Under normal conditions, the kidneys excrete about 20 mg of
ketone bodies each day, and the blood levels are maintained at about 1 mg of ketone bodies per
100 mL of blood.

In starvation, diabetes mellitus, and certain other physiological conditions in which cells do not
receive sufficient amounts of carbohydrate, the rate of ketone body formation in the liver
increases further, to a level much higher than can be used by other tissues. The excess ketone
bodies accumulate in the blood and the urine, a condition referred to as ketosis. When the
acetone in the blood reaches the lungs, its volatility causes it to be expelled in the breath. The
sweet smell of acetone, a characteristic of ketosis, is frequently noticed on the breath of severely
diabetic patients.
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Ketogenesis Pathway
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Ketosis
Ketosis is a metabolic state that your body enters when it turns fat for energy instead of
glucose. When the rate of synthesis of ketone bodies exceeds the rate of
utilization, their concentration in blood increases, this is known as
ketonemia. This is followed by ketonuria – excretion of ketone bodies in
urine. So the concept of ketonemia and ketonuria is commonly referred
to as ketosis. Smell of acetone in breathe is a common feature of
ketosis. It is most commonly associated with following conditions
Starvation: Starvation is accompanied by increased degradation of
fatty acids to meet energy needs of the body. This causes an
overproduction of acetyl-CoA which cannot be entirely handled by TCA
or citric acid cycle. Furthermore, TCA cycle is impaired due to
deficiency of oxaloacetate, since most of it is diverted for glucose
synthesis (gluconeogenesis) to meet the essential requirements (often
unsuccessful) for tissues like brain. The result is an accumulation of
acetyl-CoA and overproduction of ketone bodies leading to ketosis.
Diabetes Mellitus type I (Insulin Dependent):
1. Hyperglycaemia occurs due to decreased glucose uptake in fat and
muscle cells due to insulin deficiency
2. Lipolysis in fat cells now occurs promoted by the insulin deficiency
releasing Free fatty acids (FFA) into the blood which provide substrate
to the liver
3. A switch in hepatic lipid metabolism occurs due to the insulin
deficiency and the glucagon excess, so the excess FFA is metabolised
resulting in excess production of acetyl CoA
4. The excess hepatic acetyl CoA (remaining after saturation of TCA
cycle) is converted to ketone bodies which are released into the blood
5. Ketoacidosis and hyperglycaemia both occur due to the lack of insulin
and the increase in glucagon and most of the clinical effects follow
from these two factors

Beta oxidation of ketone bodies

The ketone bodies being water soluble are easily transported from the
liver to various tissues. Acetoacetate and β-hydroxybutyrate can be
oxidized as fuels in most tissues including skeletal muscle, brain,
certain cells of kidney and cells of the intestinal mucosa. The tissues
which lack mitochondria (eg. erythrocytes) cannot utilize ketone
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bodies. During prolonged starvation, ketone bodies are the major

sources of fuel for the brain and other parts of central nervous system
(CNS).

Steps in oxidation of ketone bodies (ketogenolysis):

1. β-hydroxybutyrate is converted back to acetoacetate.
2. Acetoacetate is activated to Acetoacetyl-CoA by a mitochondrial
enzyme thiophorase (succinyl-CoA acetoacetate-CoA transferase).
Thiophorase is absent in liver, hence ketone bodies are not utilized in
the liver.
3. Thiolase cleaves acetoacetyl-CoA to 2 moles of acetyl-CoA
4. The principal fate of the acetyl-CoA synthesized is the oxidation in TCA
cycle
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Ketogenolysis