DIGESTION AND ABSORPTION OF LIPIDS

Little or no digestion of lipids occurs in the mouth or stomach. The major site of lipid
digestion is the small intestine, where dietary lipid undergoes its major digestive processes using
enzymes secreted by pancreas.
The acidic stomach contents called chyme, containing dietary fat leaves the stomach and
enters the small intestine. Digestion of fat occurs in the duodenum by emulsification of fat, the
dietary fat is emulsified by the action of bile salts.
HYDROLYSIS OF DIETARY TRIACYLGLYCEROLS
Emulsified triacylglycerols are hydrolyzed by pancreatic lipase. Lipase hydrolyses fatty acid in
1 and 3 positions of the triacylglycerol, producing 2- monoacylglycerols and two molecules of
fatty acids
HYDROLYSIS OF DIETARY PHOSPHOLIPIDS
Dietary glycerophospholipids are digested by pancreatic phospholipase-A2. This enzyme
catalyzes the hydrolysis of fatty acid residues at the 2-position of the phospholipid, leaving
lysophospholipids and a molecule of fatty acid.
HYDROLYSIS OF CHOLESTEROL ESTER
Cholesterol esters are hydrolyzed by pancreatic cholesterol ester hydrolase (cholesterol
esterase), which produces cholesterol plus free fatty acid.
PRODUCTS OF LIPID DIGESTION (MICELLE FORMATION)
Free fatty acids, free cholesterol, 2-monoacylglycerol, and lysophospholipid are the primary
products of dietary lipid digestion. These, together with bile salts, form mixed micelles. Fat
soluble vitamins A, D, E and K are also packaged in these micelles and are absorbed from the
micelles along with the products of dietary lipid digestion.
ABSORPTION OF LIPIDS BY INTESTINAL MUCOSAL CELLS
• The mixed micelles approach the brush border membrane of the intestinal mucosal cells.
• There,the lipid components from mixed micelles are absorbed into the mucosal cells by
diffusion.
• The net result is the transfer of monoacylglycerol, fatty acids, cholesterol, and lysophospholipid
molecules into the mucosal cell.
• After absorption within the mucosal cell, the following events occur:
– 2-monoacylglycerols are reconverted to triacylglycerols.
The fatty acids absorbed from the lumen are utilized for this synthesis. Fatty acids are utilized
after its conversion to its active form, acyl- CoA.
– The absorbed lysophospholipids and cholesterol are also reconverted to phospholipids and
cholesterol esters.
– The triacylglycerol resynthesized in intestinal cells combine with cholesterol, phospholipids
and proteins to form chylomicrons.
Transport
• Triacylglycerol, phospholipid, cholesterol esters resynthesized in the intestinal mucosa and
absorbed mucosal cells into the lymph in the form of lipoprotein known as chylomicrons.
Chylomicrons are composed of:
– Triacylglycerols (85 – 90%)
– Cholesterol and cholesterol ester (5%)
– Phospholipids (7%)
– Protein (apolipoprotein B-48, 1–2%).
• The chylomicrons pass from the lymph into the blood through the thoracic duct. After a fatty
meal, the plasma is milky in appearance due to the presence of chylomicrons.

Absorption and transport of lipid from intestinal lumen where, 2-MAG: 2 Monoacylglycerol; FA: Fatty acid; C:
Cholesterol; LysoPL: Lysophospholipid; TG: Triacylglycerol; CE: Cholesterol ester

FATTY ACID OXIDATION

Fatty acids are oxidized mainly by a process called β-oxidation, in which two carbon units are
sequentially removed beginning from the carboxyl end of the fatty acid in the form of acetyl-
CoA. It is called β-oxidation because oxidation of fatty acids occurs at the β-carbon atom. β -
oxidation pathway occurs in mitochondria. It involves following three steps:
1. Activation of fatty acid to acyl-CoA
2. Transfer of acyl CoA into mitochondria by carnitine transport system
3. Reactions of β-oxidation in mitochondria

ACTIVATION OF FATTY ACID

• Before being catabolized, free fatty acids are converted to an active form called acyl-CoA. It
occurs in the cytosol in the presence of ATP, coenzyme-A (CoA-SH) and the enzyme acyl-CoA
synthetase also called thiokinase.
Subsequent steps of β-oxidation occur in the mitochondria of the liver and other tissue cells.

Transport of Acyl-CoA into Mitochondria by Carnitine Transport System

Activation of fatty acids occur in the cytosol, whereas they are oxidized in the
mitochondrial matrix. The mitochondrial inner membrane is impermeable to fatty acids. So a
special transport mechanism is needed.
Activated long chain fatty acids are carried across the inner mitochondrial membrane by
carnitine, (β-hydroxy γ-trimethyl ammonium butyrate), formed from lysine and methionine in
liver and kidney. This occurs in four steps
1. The acyl group of acyl-CoA is transferred to the carnitine to form acyl-carnitine. This reaction
is catalyzed by carnitine acyltransferase-I (CAT-I) which is located on the cytosolic face of the
inner mitochondrial membrane.
2. Acyl-carnitine is then transported across the inner mitochondrial membrane by an enzyme
translocase.
3. The acyl group is transferred back to CoA in the mitochondrial matrix by the enzyme
carnitine acyl transferase-ll (CAT-II), located on the inside of the inner mitochondrial
membrane.
4. Acyl-CoA is reformed in the mitochondrial matrix with liberation of carnitine which is
returned to the cytosolic side by the translocase in exchange for an incoming acyl-carnitine.
Reactions of β-oxidation of Fatty Acid
After the penetration of the acyl-CoA into mitochondria, it undergoes β-oxidation.
Sequence of Reactions of β-oxidation
A saturated acyl-CoA is degraded by a repeated sequence of four reactions.
1. Oxidation by FAD
2. Hydration
3. Oxidation by NAD
4. Cleavage.
1. Oxidation by FAD: The first reaction is the oxidation of acyl-CoA by an acyl-CoA
dehydrogenase to give an Δ2-trans enoyl-CoA (a trans double bond between C2 and C3). The
coenzyme for the dehydrogenase is FAD which is converted to FADH2.
2. Hydration: The next step is the hydration (addition of water) of the double bond between C2
and C3 byΔ2-enoyl-CoA hydratase to form β-hydroxy acyl-CoA.
3. Oxidation by NAD: The β-hydroxy derivative undergoes second oxidation reaction catalyzed
by β- hydroxyacyl-CoA dehydrogenase to form β-ketoacyl-CoA and generates NADH.
4. Cleavage: Finally β-ketoacyl-CoA is split at the β-carbon by thiolase to yield acetyl-CoA and
an acyl- CoA which is shorter by two carbon atoms than the original acyl-CoA that underwent
oxidation. The new acyl-CoA, containing two carbons less than the original, re-enters the β-
oxidation pathway at reaction catalyzed by acyl-CoA dehydrogenase. The process continues until
the fatty acid is degraded completely to acetyl-CoA. Acetyl-CoA can be oxidized to CO2 and
H2O via citric acid cycle in mitochondria and thus oxidation of fatty acids is completed.
β-oxidation of fatty acids
How to calculate energy yield from the β-oxidation of fatty acids
For, example,Palmitic acid has 16 carbon atoms which is activated to palmitoyl CoA
• Complete β-oxidation of palmitoyl CoA (16 carbon acid) occurs through 7 cycles of β-
oxidation yielding finally 8 acetyl-CoA, 7 FADH2 and 7 NADH.
• Three ATPs are generated when each of these NADH is oxidized by respiratory chain.
• 2 ATPs are formed for each FADH2.
• The oxidation of acetyl-CoA by the citric acid cycle yields 12 ATPs. Therefore the number of
ATPs formed in the oxidation of palmitoyl-CoA is:
– 14 ATPs from the 7 FADH2
– 21 ATPs from the 7 NADH
– 96 ATPs from the oxidation of 8 molecules of acetyl-CoA in TCA cycle.
– Total of 131 ATPs.
• Two high energy phosphate bonds are consumed in the activation of palmitate, in which ATP is
split into AMP and PPi.
• Thus, the net yield from the complete oxidation of palmitic acid (palmitate) is 129 ATPs.

REGULATION OF β-OXIDATION
• Rate limiting step in the β-oxidation pathway is the formation of acyl-carnitine which is
catalyzed by carnitine-acyl transferase-I (CAT-I). CAT-I is an allosteric enzyme. Malonyl-
CoA is an inhibitor of CAT-I.
– In well-fed state due to increased level of insulin, concentration of malonyl-CoA increases
which inhibits CAT-I and leads to decrease in fatty acid oxidation.
– In starvation, due to increased level of glucagon concentration of malonyl-CoA decreases and
stimulates the fatty-acid oxidation.
β-OXIDATION OF A FATTY ACID WITH AN ODD NUMBER OF CARBON ATOMS
• Fatty acids, having an odd number of carbon atoms, are oxidized by β-oxidation in the same
way as fatty acids, having an even number, except in the last and final β-oxidation cycle of odd
carbon fatty acids a three carbon fragment propionyl CoA is formed rather than two carbon
fragment acetyl-CoA.
• The Propionyl-CoA is then converted to succinyl-CoA, a constituent of citric acid cycle
through the reaction sequence below
• Propionyl-CoA is carboxylated at the expense of an ATP to yield D-methylmalonyl-CoA;
catalyzed by propionyl-CoA carboxylase, a biotin enzyme.
• The D-methylmalonyl-CoA is converted to the L-methylmalonyl-CoA by the enzyme
methylmalonyl- CoA epimerase.
• Succinyl-CoA is formed from L-methylmalonyl-CoA by methylmalonyl-CoA mutase, which
requires vitamin B12 as a coenzyme.
PEROXISOMAL FATTY ACID OXIDATION
Oxidation of very long chain fatty acids, which contain 20–26 carbons begins in peroxisomes by
a process very similar to β-oxidation (and is completed in the mitochondria) with significant
differences.
• The action of first enzyme acyl-CoA dehydrogenase differs, in that, it produces H2O2 rather
than FADH2. In peroxisomes the FADH2 is not linked to ETC for generation of ATP but it
transfers reducing equivalents directly to O2, yielding H2O2.That is, there is no ATP synthesized
from peroxisomal β-oxidation of fatty acids.The H2O2 produced is converted to water and
molecular oxygen the enzyme, Catalase.

DISORDER OF PEROXISOMAL FATTY ACID OXIDATION

Zellweger syndrome
• A rare inborn error of peroxisomal fatty acid oxidation is due to inherited absence of
functional peroxisomes in all tissues. As a result, the long chain fatty acids are not oxidized in
peroxisomes and accumulate in tissues, particularly in brain, liver, kidney and muscle and
usually results in death by age six.
α-oxidation
• Oxidation occurs at α-carbon of fatty acid in which one carbon is removed from the carboxyl
end of the fatty acid chain and released as CO2.The remaining carbons of the fatty acid can repeat
the process. It does not require CoA intermediates and does not generate ATP.This process
occurrs in brain and other nervous tissues.
DISORDERS ASSOCIATED WITH IMPAIRMENT OF α-FATTY ACID OXIDATION
Refsum’s disease (phytanate storage disease)
A rare neurologic inborn error of lipid metabolism results from a genetic deficiency in phytanic
acid α-hydroxylase required for the hydroxylation of phytanic acid by α-oxidation. Persons with
Refsum’s disease have an inherited defect in α-oxidation that leads to accumulation of phytanic
acid in the nerve tissue.Clinical symptoms include retinitis pigmentosa (progressive
dengeneration of retina), peripheral neuropathy and ataxia. Treatment involves elimination of
dietary sources of phytol.
ω-oxidation
Fatty acids may be, oxidized at the ω-carbon of the chain by enzymes in the endoplasmic
reticulum. ω-carbon is a carbon farthest from the carboxyl end.The ω-methyl group is first
oxidized to an alcohol (CH2OH) by hydroxylase that uses cytochrome P450, molecular oxygen
and NADPH in the endoplasmic reticulum.
• The alcoholic group is subsequently oxidized to –COOH by dehydrogenase and thus forming a
dicarboxylic acid.
• Short chain dicarboxylic acid such as pimelic acid, a precursor of biotin is formed by ω-
oxidation.

METABOLISM OF KETONE BODIES

Acetoacetate, β-hydroxybutyrate and acetone, these three substances are collectively known as
ketone bodies. These are water soluble and energy yielding

KETOGENESIS
Ketogenesis means the formation of ketone bodies. Liver is the only organ that synthesizes
ketone bodies. The synthesis of ketone bodies occurs in mitochondrial matrix of hepatic cells.
Ketogenesis occurs by the following reactions:
• Two molecules of acetyl-CoA condense to form acetoacetyl-CoA. This reaction is catalysed
by thiolase.
• Acetoacetyl-CoA then condenses with one more molecule of acetyl-CoA to give β-hydroxy-β
methylglutaryl- CoA (HMG-CoA), catalysed by HMG-CoA synthase.
• β-HMG-CoA is then cleaved to acetyl-CoA and acetoacetate by the enzyme HMG-CoA lyase
present in the mitochondria.
• β-hydroxy butyrate is formed by the reduction of acetoacetate in the mitochondrial matrix by
the enzyme β-Hydroxybutyrate dehydrogenase.
• Acetoacetate also undergoes a slow, nonenzymatic spontaneous decarboxylation to acetone.
• The concentration of total ketone bodies in the blood of well-fed condition does not normally
exceed 0.2 mmol/L
UTILIZATION OF KETONE BODIES
The site of production of ketone bodies is the liver. But the liver cannot utilize ketone bodies
because it lacks the particular enzyme CoA-transferase which is required for the activation of
ketone bodies. Acetoacetate, β-hydroxybutyrate and acetone diffuse from the liver mitochondria
into the blood and are transported to peripheral tissues.
• In peripheral tissues the β-hydroxybutyrate is first reconverted to acetoacetate and the
acetoacetate is then reactivated to acetoacetyl-CoA.
• Activation of acetoacetate occur by CoAtransferase in the presence of succinyl-CoA.
Acetoacetate is activated by the transfer of CoA from succinyl-CoA.
• Acetoacetyl-CoA, formed by this reaction, is then cleaved by thiolase to yield two molecules of
acetyl- CoA which can be oxidized in the citric acid cycle to H2O and CO2.
• Further metabolism of acetone does not occur. Because acetone is volatile, it is expired by the
lungs. Schematic representation of overall metabolism (synthesis and breakdown) of ketone
bodies is shown below

Regulation of Ketogenesis
The ketone body formation is regulated at the following three important levels.
1. Factors regulating lipolysis: Free fatty acids the precursors of ketone bodies arise from
lipolysis of triacylglycerol in adipose tissue. So factors regulating lipolysis also regulate
ketogenesis. The hormone glucagon stimulates lipolysis which in turn stimulates ketogenesis,
whereas insulin inhibits lipolysis and ketogenesis.
2. Factors regulating β-oxidation of fatty acids
• Carnitin acyl transferase I (CAT I) regulates the fatty acid oxidation which, in turn, regulates
ketogenesis as well. Its activity is high in starvation leading to increased fatty acid oxidation and
formation of ketone bodies.
3. Factors regulating oxidation of acetyl-CoA: In turn, the acetyl-CoA, formed in β-oxidation
in liver mitochondria, has two possible fate.

• It may be oxidized to CO2 via the citric acid cycle or it enters the pathway of ketogenesis to
form ketone bodies.
• When oxaloacetate concentration is very low, little acetyl-CoA enters the citric acid pathway,
and ketone body formation is then favoured.
• Concentration of oxaloacetate is lowered if carbohydrate is unavailable or improperly utilized,
e.g. in fasting or in diabetes. Under these conditions, acetyl-CoA is diverted to the formation of
acetoacetate.
DISORDERS OF KETONE BODY METABOLISM
Ketosis
• Normally the concentration of ketone bodies in blood is very low, (less than 0.2 mmol/L) but in
fasting and in diabetes mellitus it may reach extremely high levels.
• During fasting or in diabetes mellitus, when exogenous glucose is unavailable, insulin secretion
is inhibited. The plasma insulin concentration is, therefore, low which causes increased lipolysis
and therefore increased production of free fatty acids and hence ketone bodies production is
increased.
• When the rate of formation of the ketone bodies by liver exceeds the capacity of the peripheral
tissues to use them up, their levels begin to rise in blood.
An increase in concentration of ketone bodies in blood is called ketonemia and eventually leads
to excretion of ketone bodies into the urine called ketonuria. The overall condition (ketonemia
and ketonuria) is called ketosis.
• In addition to β-hydroxybutyrate and acetoacetate, the blood of diabetics also contains acetone.
Acetone is very volatile and is present in the breath of diabetics, to which it gives sweet fruity
odour.

KETOACIDOSIS
• Since acetoacetate and β-hydroxybutyrate are moderately strong acids, increased levels of these
ketone bodies decrease the pH of the blood and cause metabolic acidosis. The acidosis caused
by over production of ketone bodies is termed as ketoacidosis.
• Acetoacetate and β-hydroxybutyrate acids when present in high concentration in blood, are
buffered by HCO3.The excessive use of HCO3 – depletes the alkali reserve causing
ketoacidosis.
• Ketoacidosis is seen in type I diabetes mellitus, whereas in type II diabetes ketoacidosis is
relatively rare.

DE NOVO SYNTHESIS OF FATTY ACIDS (LIPOGENESIS)

In humans, fatty acid synthesis occurs mainly in the liver and lactating mammary glands
and, to a lesser extent, in adipose tissue, kidney and brain. De novo synthesis means new
synthesis. Fatty acid synthesis occurs in three phases:
1. Transport of acetyl-CoA from mitochondria to cytosol.
2. Carboxylation of acetyl-CoA to malonyl-CoA.
3. Reactions of fatty acid synthase complex.
TRANSPORT OF ACETYL-COA FROM MITOCHONDRIA TO CYTOSOL
Fatty acids are synthesized in the cytosol, whereas acetyl-CoA is formed in mitochondria, hence,
acetyl-CoA must be transferred from mitochondria to the cytosol. Since acetyl CoA is
impermeable to inner mitochondrial membrane, it is transferred in the form of citrate from
mitochondrial matrix to the cytosol as shown below
• Citrate is formed in the mitochondrial matrix by the condensation of acetyl-CoA with
oxaloacetate (first reaction in the citric acid cycle).
• Then citrate is transported to the cytosol by translocase, where it is cleaved by citrate lyase to
oxaloacetate and acetyl-CoA.
• Oxaloacetate formed in this reaction must be returned to the mitochondria. The inner
mitochondrial membrane is impermeable to oxaloacetate.
Therefore, oxaloacetate is reduced to malate, catalyzed by cytosolic malate dehydrogenase.
Then
malate is oxidatively decarboxylated to pyruvate by NADP-linked malic enzyme. NADPH and
CO2 are generated in this reaction. Both of them are utilized for fatty acid synthesis.
• The pyruvate formed in this reaction readily diffuses into mitochondria, where it is
carboxylated to oxaloacetate by pyruvate carboxylase.

CARBOXYLATION OF ACETYL-COA TO MALONYL-COA

• Carboxylation of acetyl-CoA to malonyl-CoA is the initial and rate limiting reaction in fatty
acid synthesis.
• This reaction is catalysed by an enzyme complex, acetyl-CoA carboxylase that contains biotin
and utilizes bicarbonate (as a source of CO2) in presence of ATP.

REACTIONS OF FATTY ACID SYNTHASE COMPLEX

• Once malonyl CoA is formed from acetyl CoA, the De novo synthesis of palmitic acid is
carried out in cytosol by fatty acid synthase complex.
• Fatty acid synthase complex is a multienzyme complex possessing 7 different enzymes and one
acyl carrier protein (ACP) molecule.
The fatty acid synthase complex has two active –SH groups. One -SH group is of 4-
phosphopantetheine moiety of ACP. Another -SH group is of cysteine residue of enzyme
ketoacyl synthase. Both –SH groups participate in fatty acid biosynthesis.
• Fatty acid synthase complex is a dimer.
Each monomer is identical consisting of all seven enzyme activities of fatty acid synthase and an
ACP.
• The two subunits associate in a head-to-tail arrangement, so that the phosphopantetheine-SH
group of one subunit and a cysteinyl-SH group of another subunit are closely aligned.
• Though each monomer contains all the enzyme activities but only the dimer is functionally
active.
This is because the functional unit consists of half of each subunit interacting with the
complementary half of the other. The two functional subunits of fatty acid synthase can
synthesize two fatty acids simultaneously.
The fatty acid synthase complex catalyses the following reactions
• In the first reaction, catalyzed by acetyl transacylase the acetyl group of acetyl-CoA is
transferred to the cysteine -SH group of the 3-ketoacyl synthase of fatty acid synthase complex
(designated E).
• In the second reaction, the malonyl group of malonyl-CoA is transferred to the
phosphopantetheine –SH group (Pan-SH) of ACP catalysed by malonyl transacylase to form
acetyl-malonyl enzyme.
• The acetyl and malonyl groups, covalently bonded to -SH groups of the synthase, undergo a
condensation reaction. This reaction is catalysed by 3-ketoacyl synthase. The acetyl group
attached to cysteine -SH is transferred to malonyl group bound to -SH pantetheine.
Simultaneously the malonyl moiety loses CO2 forming 3-ketoacyl enzyme.
• The 3-ketoacyl enzyme undergoes reduction at the 3-keto group, at the expense of NADPH as
electron donor to form 3-Hydroxyacyl enzyme, catalysed by 3-ketoacyl reductase.
• 3-hydroxyacyl enzyme is dehydrated by 3-hydroxyacyl hydratase to yield 2, 3 unsaturated acyl
enzyme.
• 2,3 unsaturated acyl enzyme is reduced to form acyl enzyme containing 4-carbon, by the action
of enoyl reductase and NADPH is the electron donor.
• This acyl group (4 carbon) is now transferred from pantetheine -SH group to the cysteine -SH
group.
• To lengthen the chain by another 2-carbon unit, the sequence of reactions is repeated, a new
malonyl residue being incorporated during each sequence, until a saturated 16-carbon acyl
radical (palmityl) has been assembled.
• Thus, after a total of seven such cycles, palmitoyl enzyme is formed, which is liberated from
the enzyme complex by the activity of a seventh enzyme in the complex thioesterase
(deacylase).
Schematic diagram of fatty acid synthase multienzyme complex

Regulation of Fatty Acid Synthesis

De novo fatty acid synthesis is regulated by Acetyl-CoA Carboxylase
• Acetyl-CoA carboxylase which catalyses the formation of malonyl-CoA is the rate limiting
enzyme in the fatty acid synthesis. It is regulated in the following ways:
1. Allosteric regulation: Acetyl-CoA carboxylase is allosterically activated by citrate and
inhibited by its own product palmitoyl-CoA.
2. Hormonal regulation: The activity of acetyl- CoA carboxylase is also controlled by
hormones.
Glucagon and epinephrine inactivate the enzyme. In contrast, insulin activates the enzyme.
Thus, fatty acid synthesis is stimulated by insulin and inhibited by glucagon and epinephrine.
3. Nutritional regulation: Synthesis of acetyl-CoA carboxylase is increased with high
carbohydrate and low fat diet, which promote fatty acid synthesis. By contrast, synthesis of
acetyl-CoA carboxylase decreases during starvation, diabetes mellitus or high fat diet.

SYNTHESIS OF LONG CHAIN FATTY ACIDS FROM PALMITATE

The major product of fatty acid synthesis is palmitate (palmitic acid). Longer fatty acids are
formed by elongation reactions either in endoplasmic reticulum (microsomes) or in
mitochondria.
Microsomal elongation of fatty acids
The microsomal enzyme fatty acid elongase elongates palmitate by the addition of 2-carbon
fragments derived from malonyl-CoA and NADPH provides the reducing equivalents. The
elongation reaction resembles those of fatty acid synthesis, except that the fatty acyl chain is
attached to coenzyme-A rather than to phosphopantetheine group of ACP.
Mitochondrial elongation of fatty acids
Fatty acids can be elongated in mitochondria, but mitochondrial elongation of fatty acids is less
active. In this case, the source of the 2-carbon units is acetyl- CoA and the substrates are usually
fatty acids containing less than 16-carbons, mainly short and medium chain fatty acids.
TRIACYLGLYCEROL METABOLISM
Triacylglycerols are esters of the glycerol (alcohol) and fatty acids. Fatty acids derived from
endogenous synthesis or from the diet are stored in the adipose tissue in the form of
triacylglycerol, called “neutral fat.” Triacylglycerol serves as the body’s major fuel storage
reserve.

BIOSYNTHESIS OF TRIACYLGLYCEROLS
Triacylglycerols are synthesized in both liver and adipose tissue
• First fatty acids are activated to acyl-CoA
• Then two molecules of acyl-CoA combine with glycerol-3-phosphate to form phosphatidic acid
(1,2- diacylglycerol phosphate) via formation of lysophosphatidic acid.
• Dephosphorylation of phosphatidic acid produces diacylglycerol.
• A further molecule of acyl-CoA is esterified with diacylglycerol to form triacylglycerol.
• The sources of glycerol-3-phosphate, which provides the glycerol moiety for triacylglycerol
synthesis, differ in liver and adipose tissue.
• In liver glycerol-3-phosphate is produced from the phosphorylation of glycerol by glycerol
kinase or from the reduction of dihydroxyacetone phosphate (DHAP), derived from glycolysis.
• Adipose tissue lacks glycerol kinase and can produce glycerol-3-phosphate only from glucose
via dihydroxyacetone phosphate (DHAP).
Fate of Triacylglycerol Formed in Liver and Adipose Tissue
The fate of triacylglycerol in liver and adipose tissue is different.
• In the liver, little triacylglycerol is stored; instead most is exported in the form of very low
density lipoprotein (VLDL). Once released into the blood stream, triacylglycerol of VLDL is
hydrolyzed by lipoprotein lipase, which is located on the walls of blood capillaries. It clears the
triacylglycerol in VLDL, forming free fatty acids and glycerol.
• In adipose tissue triacylglycerol is stored in the cells. It serves as “depot fat” ready for
mobilization when the body requires it for fuel.

ADIPOSE TISSUE METABOLISM

Triacylglycerol is the major storage form of lipid in humans. Adipose tissue is the main store of
triacylglycerol in the body. The triacylglycerol stored in adipose tissue is continually undergoing
lipolysis (hydrolysis) and resynthesis.

Synthesis of Triacylglycerol in Adipose Tissue

• In adipose tissue triacylglycerol is synthesized from acyl-CoA and glycerol-3-phosphate.
• For provision of glycerol-3-phosphate, the tissue is dependent on glycolysis and a supply of
glucose.
The transport of glucose into adipose tissue is stimulated by insulin.
• Some fatty acids that are incorporated into the triacylglycerol are synthesized within the
adipose tissue from glucose. The remainder is taken up from the blood, which is formed by the
action of lipoprotein lipase on the triacylglycerol of chylomicrons and VLDL. Insulin stimulates
this process by stimulating lipoprotein lipase.
DEGRADATION OF TRIACYLGLYCEROLS IN ADIPOSE TISSUE
To leave the adipose tissue, the triacylglycerol must be hydrolyzed to fatty acids and glycerol.
• Triacylglycerol undergoes hydrolysis by a hormone sensitive lipase to form free fatty acids and
glycerol.
• This lipase is distinct from lipoprotein lipase that catalyses hydrolysis of triacylglycerol before
its uptake into extrahepatic tissues.
• The glycerol produced by lipolysis cannot be used by adipose tissues because they lack the
enzyme glycerol kinase.
• Therefore, the glycerol is transported to the liver which contains, glycerol kinase to metabolize
glycerol to glucose by the process of gluconeogenesis.
• The free fatty acid formed by lipolysis can be reconverted in the adipose tissue to acyl-CoA by
acyl-CoA synthetase and re-esterified with glycerol-3- phosphate to form triacylglycerol.
Thus, there is a continuous cycle of lipolysis and resynthesis (re-esterification) within the
adipose tissue.
However, when the rate of re-esterification is not sufficient to match the rate of lipolysis, free
fatty acids accumulate and diffuse into the plasma and raise the concentration of plasma free
fatty acids, which occurs during fasting, diabetes mellitus, anxiety or physical exertion.
Adipose tissue metabolism where, PPP: Pentose-phosphate pathway; TG: Triacylglycerol; FFA:
Free fatty acid; DHAP: Dihydroxy acetone phosphate

Regulation of Adipose Tissue Metabolism

• High levels of insulin in the blood stimulate triacylglycerol formation in adipose tissue and
inhibits the lipolysis by inhibiting hormone sensitive lipase.
• Conversely, low insulin concentrations enhance the mobilization of fatty acid from adipose
tissue
• Other hormones like epinephrine and glucagon accelerate the rate of lipolysis of triacylglycerol
stores. These hormones activate hormone sensitive lipase.

FATTY LIVER
Fatty liver is the excessive accumulation of fat primarily neutral fat, triacylglycerol in the liver.
Fat is mainly stored in adipose tissue. Liver is not a storage organ. It contains about 5% fat. In
pathological conditions, this may go up to 25–30% and is known as fatty liver or fatty
infiltration of liver. When accumulation of lipid in the liver becomes chronic, fibrotic changes
occur in cells which may finally lead to cirrhosis and impairment of liver function.
Causes of Fatty Liver
Fatty liver may occur due to:
1. Overproduction of triacylglycerol in liver.
2. Impaired synthesis of VLDL.
• Overproduction of triacylglycerol: Fatty liver is associated with increased level of plasma
free fatty acids from the diet (high fat diet) or from the adipose tissue during starvation, diabetes
mellitus and alcoholism. The increasing amounts of fatty acids are taken up by the liver and
esterified to triacylglycerol. VLDL produced in liver carries this triacylglycerol from liver to the
peripheral tissues. But the production of VLDL does not keep pace with the formation of
triacylglycerol, allowing triacylglycerol to accumulate in liver.
• Impaired synthesis of VLDL: Impairment in the biosynthesis of VLDL, in turn, impairs the
transport of triacylglycerol from liver, thus allowing triacylglycerol to accumulate in the liver.
The defect in the synthesis of VLDL may be due to:
1. A block in apolipoprotein synthesis
2. Defect in the synthesis of phospholipids that are found in lipoproteins.
3. A failure in the formation mechanism of lipoprotein itself from lipid and apoprotein.

FACTORS THAT CAUSE FATTY LIVER

1. High fat diet: Due to increased supply of free fatty acids from the diet, capacity of liver for
lipoprotein formation is outweighed.
2. Starvation or uncontrolled diabetes mellitus or insulin insufficiency: Due to increased
mobilization of free fatty acids from adipose tissue.
3. Alcoholism: Due to increased hepatic triacylglycerol synthesis and decreased fatty acid
oxidation.
4. Dietary deficiency of:
i. Lipotropic factors: Deficiency of lipotropic factors like choline, betaine, methionine and
lecithin may cause fatty liver. Choline is required for the formation of phospholipid lecithin,
which in turn, is an essential component of lipoprotein. Betain and methionine possessing methyl
groups can be used to synthesize choline.
ii. Essential fatty acids: Essential fatty acids are required for the formation of phospholipid. A
deficiency of essential fatty acids leads to decreased formation of phospholipids.
iii. Essential amino acids: Essential amino acids are required for the formation of
apolipoprotein and lipotropic factor choline.
iv. Vitamin E and selenium: Deficiency of vitamin E or selenium enhances the hepatic
necrosis. They have protective effect against fatty liver.
v. Protein deficiency: For example, in kwashiorkor, deficiency of protein impairs formation of
apolipoprotein.
vi. Vitamin deficiency: Deficiency of pyridoxine and pantothenic acid decrease the availability
of ATP, needed for protein biosynthesis.
5. High cholesterol diet: Excess amount of cholesterol in diet competes for essential fatty acids
for esterification.
6. Use of certain chemicals: For example, puromycin, chloroform, carbon tetrachloride, lead
and arsenic inhibit protein biosynthesis and impair formation of apolipoprotein.

Lipotropic Factors
• The substances that prevent the accumulation of fat in the liver are known as lipotropic factors.
Dietary deficiency of these factors can result in fatty liver. The various lipotropic agents are:
– choline
– methionine
– betaine, etc.
• Choline is the principal lipotropic factor, and other lipotropic agents act by producing choline
in the body, e.g. betaine and methionine possessing methyl groups that are donated to
ethanolamine to form choline.
• Choline is required for the formation of phospholipid, lecithin, which in turn, is an essential
component of lipoprotein.
• Vitamin B12 and folic acid are also able to produce lipotropic effect, as these are involved in
the formation of methionine from homocysteine.
• Casein and other proteins also possess lipotropic activity.

CHOLESTEROL METABOLISM
Cholesterol is the major sterol in human and has cyclopentanoperhydrophenanthrene ring
system as a parent structure.
Cholesterol is an amphipathic lipid which can be synthesized by most cells of the body and it is
obtained from the diet in foods of animal origin. It is not synthesized in plants. The major
sources of dietary cholesterol are egg yolk and meat, particularly liver.
De Novo Synthesis of Cholesterol
Cholesterol is synthesized by a pathway that occurs in most cells of the body. Liver and intestine
are major sites of cholesterol synthesis.
All 27-carbon atoms of cholesterol are derived from the acetyl-CoA. The enzyme system of
cholesterol synthesis are present in cytosolic and microsomal (endoplasmic reticulum) fractions.
The reactions of cholesterol biosynthesis occurs into 5 stages

Five stages of cholesterol biosynthesis

Biosynthesis of cholesterol, showing its five stages
Stage 1: Synthesis of Mevalonate from Acetyl-CoA through HMG-CoA
• First, two molecules of acetyl-CoA condense to form acetoacetyl-CoA, catalyzed by a
cytosolic
thiolase enzyme.
• Next, a third molecule of acetyl-CoA condenses with acetoacetyl-CoA catalyzed by HMG-
CoA synthase to form HMG-CoA.
• This sequence of reactions in the cholesterol synthesis is similar to those for the synthesis of
ketone bodies except that ketone body synthesis occurs in mitochondria and cholesterol is in the
cytosol.
• The next step is catalyzed by HMG-CoA reductase. This enzyme uses two molecules of
NADPH and converts HMG-CoA to mevalonate. This is the rate limiting step in the cholesterol
synthesis.

Stage 2: Formation of Isoprenoid Unit by Decarboxylation of Mevalonate

Mevalonate is phosphorylated by ATP and subsequently decarboxylated to form a five carbon
isoprene unit, isopentenyl pyrophosphate (IPP).
Stage 3: Formation of squalene from condensation of six isoprenoid units
Six isopentenyl pyrophosphate molecules condense with loss of their pyrophosphate groups to
yield the squalene (30-carbon atoms compound) through the formation of geranyl pyrophosphate
and farnesyl pyrophosphate.
Stage 4: Cyclization of squalene to lanosterol
Squalene undergoes a series of complex enzymatic reactions, in which its linear structure is
folded and cyclized to form lanosterol, which has the four condensed rings that form the steroid
nucleus of cholesterol.
Stage 5: Formation of cholesterol from Lanosterol
The conversion of lanosterol to cholesterol is a multistep process, resulting in the
• Shortening of the carbon chain from 30 to 27
• Removal of the three methyl groups at C4
• Migration of the double bond from C8 to C5
• Reduction of the one double bond between C24 and C25 by NADPH.

REGULATION OF DE NOVO SYNTHESIS OF CHOLESTEROL

HMG-CoA reductase is the rate limiting enzyme in cholesterol biosynthesis and is subjected to
different kinds of metabolic control. The following are the different kinds of metabolic control.
Feedback Regulation
• Cholesterol the end product of the pathway, as well as mevalonate, repress the synthesis of
HMG-CoA reductase.
• It is also repressed by bile salts or bile acids, thus decreasing the cholesterol synthesis.
Hormonal Regulation
• Cholesterol synthesis is increased by insulin and thyroid hormone and is decreased by
glucagon and glucocorticoid by stimulating and inhibiting HMG-CoA reductase respectively.
Nutritional Regulation
• Dietary cholesterol suppresses the synthesis of the HMG-CoA reductase in the liver.
• Increased caloric intake stimulates cholesterol synthesis primarily by increasing the availability
of acetyl-CoA and NADPH
Drugs like mevastatin and lovastatin inhibit cholesterol synthesis by acting as competitive
inhibitors of HMG-CoA reductase. These drugs are used to decrease the serum cholesterol level
in patients having elevated serum cholesterol concentration.

TRANSPORT OF CHOLESTEROL
Transport of Dietary Cholesterol from Intestine
• After digestion, dietary cholesterol is absorbed into the intestinal mucosal cells, where much of
it is subsequently reconverted into cholesterol esters.
• Cholesterol esters that are synthesized in the mucosal cells, together with some unesterified
cholesterol are incorporated into chylomicrons, which transports cholesterol and other dietary
lipids from intestine.
• When triacylglycerol of chylomicrons hydrolysed by lipoprotein lipase in the peripheral
tissue, only about 5% of the cholesterol ester is lost. The rest is taken up by the liver in the form
of chylomicron remnants and is hydrolysed to cholesterol.
Transport of Cholesterol from Liver to Peripheral Tissue
• Cholesterol from liver is transported in the form of VLDL into the plasma.
• Like chylomicrons VLDL triacylglycerol is hydrolyzed by lipoprotein lipase in the peripheral
tissues and results with formation of cholesterol rich LDL.
• LDL cholesterol is taken up by the liver or extrahepatic tissues by LDL receptors.

Transport of cholesterol where, C:Cholesterol; LPL:Lipoprotein lipase

Transport of Cholesterol from Peripheral Tissue to Liver
• HDL picks up cholesterol from the peripheral tissues and converts it to cholesterol esters by
LCAT enzyme
• These cholesterol esters are ultimately returned to the liver and thus HDL is said to participate
in “reverse cholesterol transport.”
Degradation of Cholesterol
Cholesterol can undergo degradative reactions in humans with conversion of cholesterol to
physiologically important products such as:
• Bile acids which is the main catabolic pathway
• Steroid hormones
• Vitamin D

Formation of Bile Acids

The primary bile acids cholic acid and chenodeoxycholic acid are synthesized in the liver from
cholesterol

Synthesis of bile acid and its regulation

IMPORTANCE OF BILE ACIDS
• The bile acids required for the emulsification of the dietary lipids and facilitate the enzymatic
digestion and absorption of dietary lipids.
• Conversion of cholesterol into bile acid in the liver prevents the body from becoming
overloaded with cholesterol.
• As steroidal ring of cholesterol cannot be degraded in the body, the excretion of bile salts
serves as a major route for removal of the steroid ring from the body.
Formation of Steroid Hormones
Cholesterol is the precursor of the five classes of steroid hormones:
1. Progesterone
2. Glucocorticoids
3. Mineralocorticoids
4. Sex hormones, androgens
5. Estrogens.
Formation of Vitamin D
Cholesterol is also the precursor of vitamin D which regulates calcium and phosphorus
metabolism.

The formation of vitamin D3 from cholesterol

Excretion of Cholesterol
• Cholesterol is excreted in faeces
• Unlike many other metabolites, cholesterol cannot be destroyed by oxidation to CO 2 and H2O,
because of absence of enzymes capable of catabolizing the steroid ring.
• It is excreted in the bile either as cholesterol or after conversion to bile acids. About 1 gm of
cholesterol is eliminated from the body per day. Roughly, half is excreted in the form of bile
acids and half is in the form of cholesterol.
• Moreover, some dietary cholesterol is excreted in faeces without being absorbed.
• Some of the cholesterol in the intestine is acted on by intestinal bacterial enzymes and
converted to neutral sterols, coprostanol, cholestanol and excreted through faeces.
Blood Cholesterol
The normal total plasma cholesterol lies between 150–250 mg per 100 ml. Cholesterol present
in blood occurs both free and as ester form. The hydroxyl group at position three of cholesterol
can be esterified with fatty acids producing cholesterol esters. About 70% of the plasma
cholesterol is esterified with fatty acids to form cholesterol ester, remaining 30% is free
cholesterol.
GOOD CHOLESTEROL AND BAD CHOLESTEROL
• HDL cholesterol is considered to be the good cholesterol, because it removes excess cholesterol
from peripheral tissues and transport it to the liver where it is degraded or excreted in the bile.
HDL thus tends to lower blood cholesterol level.
• On the other hand LDL cholesterol is called bad cholesterol, because it transports cholesterol
from liver to the peripheral tissue. Whereas for excretion, cholesterol must enter the liver. The
risk of coronary heart disease (CHD) is related to plasma levels of total cholesterol and LDL-
cholesterol.
Hypercholesterolemia
In a normal adult, the total plasma cholesterol ranges form 150–250 mg/100 ml. An increase in
plasma cholesterol more than 250 mg/100 ml is known as hypercholesterolemia and is seen in
the following conditions:
1. Diabetes mellitus: This is due to deficiency of insulin, rate of lipolysis is increased. Increased
rate of lipolysis results in more release of free fatty acids in circulation which increases acetyl
CoA. Excess of acetyl-CoA are diverted for cholesterol biosynthesis.
2. Hypothyroidism: This is due to decrease in the HDL receptors on hepatocyte in
hypothyroidism and due to decreased synthesis of 7-α-hydroxylase that requires for the
conversion of cholesterol to bile acids. Thyroid hormone induces the synthesis of 7-α-
hydroxylase.
3. Obstructive jaundice: Due to decreased excretion of cholesterol through bile.
4. Familial hypercholesterolemia: It is a genetic disease caused by deficiency or malfunction of
the LDL receptors. In the absence of these receptors, the liver cannot take LDL and there is no
release of cholesterol of LDL into the liver. Therefore, they do not cause feedback inhibition of
cholesterol synthesis and lead to increased cholesterol formation.

ATHEROSCLEROSIS
High level of serum cholesterol results in atherosclerosis. The atherosclerosis is characterized
by hardening and narrowing of the arteries due to deposition of cholesterol and other lipids in the
inner arterial wall. Deposition of cholesterol and other lipids in the inner arterial wall leads to
formation of plaque (sticky deposit) and results in the endothelial damage and narrowing of the
arterial lumen. The hardening and narrowing of coronary arteries results in coronary heart
disease (CHD).
Factors responsible for development of atherosclerosis
Age
Aging brings about changes in the blood vessel wall due to decreased metabolism of cholesterol.
As age advances, the elasticity of the vessel wall decreases and formation of plaques progresses.
The plaques are composed of smooth muscle cells, connective tissue, lipids and debris that
accumulate in the inner side of the arterial wall. Formation of plaque leads to narrowing of the
lumen and invites atherosclerosis.
Sex
Males are affected more than females, female incidence increases after menopause. Suggesting
that male sex hormone might be atherogenic or conversely that female sex hormones might be
protective.
Genetic factor
Hereditary genetic derangement of lipoprotein metabolism leads to high blood lipid level and
familial hypercholesterolemia.
Hyperlipidemia
Increased levels of the following components of plasma lipids are associated with increased risk
of atherosclerosis
• Total serum cholesterol and triacylglycerol.
• Low density lipoprotein (LDL) is richest in cholesterol and deposits it in tissues and has
maximum association with atherosclerosis.
• Lipoprotein(a) (LPa): Some people have a special type of abnormal LDL called LP(a)
containing an additional protein, apoprotein-a. Elevated LPa levels are associated with an
increased risk of coronary heart disease.
Level of HDL
Low level of HDL is associated with atherosclerosis. HDL has protective effect against
atherosclerosis. HDL participates in reverse transport of cholesterol, i.e. it transports cholesterol
from cells to the liver for excretion in the bile. The higher the levels of HDL, the lower is the risk
of ischemic heart disease. Consequently, high ratio of HDL/LDL reduces the development of
atherosclerosis. There is an inverse relationship between cardiovascular risks and HDL
concentration.
Hypertension
Hypertension is the major risk factor in patients over 45 years of age. It acts probably by
mechanical injury of the arterial wall due to increased blood pressure.
Cigarette smoking
Ten cigarettes per day increase the risk three-fold due to reduced level of HDL and accumulating
carbon monoxide that may cause endothelial cell injury. Cessation of smoking decreases risk to
normal after 1 year.
Diabetes mellitus
The risk is due to the coexistence of other risk factors such as obesity, hypertension, and
hyperlipidemia.
Minor or soft risk factors
These include lack of exercise, stress, obesity, high caloric intake, diet containing large
quantities of saturated fats, use of oral contraceptive, alcoholism and hyperuricemia, etc. The risk
is due to increased LDL and decreased HDL levels.
Prevention of atherosclerosis
• The most important preventive measure against the development of atherosclerosis is to eat a
low fat diet that contains mainly unsaturated fat with low cholesterol content.
• Natural antioxidants such as vitamin E, C or β-carotene may decrease the risk of cardiovascular
disease by protecting LDL against oxidation.
• Moderate consumption of alcohol and exercise appears to have a slightly beneficial effect by
raising the level of HDL.
• Drug therapy, e.g. Lovastatin, clofibrate, cholestyramine which inhibit cholesterol synthesis.

OBESITY
Obesity is the most prevalent nutritional disorder in prosperous developed countries and among
affluent people in developing and underdeveloped countries.Obesity is the condition in which
excess fat has accumulated. This is due to the increased energy intake and decreased energy
expenditure. The obesity index is calculated as W/H2 (where W = weight (kg) and H = height
(m)
"Obesity index" is an old terminology; the modern expression is Body Mass Index (BMI).
The degree of obesity is commonly assessed by means of the body mass index (BMI) .
Body weight (kg)
BMI = —————————
Height (m2)

Relationship between BMI and degree of obesity

Value of BMI Degree of obesity
20-25 Normal( not obese)
25-30 Overweight or obesity grade I
30-35 Over obesity or grade II
above 35 Gross obesity or grade III

Generally, a person is obese when BMI exceeds 27.8 kg/m 2 in men and 27.3 kg/m2 in women
(excess of 120% of desirable body weight).
Obesity can occur only as a result of ingestion of food in excess of the body's needs. The
major causes are food habits (intake of calorie rich food in excess amounts) and lack of exercise.
There is an increase in the number (hyperplasia) and size (hypertrophy) of the adipocytes.
Genetic predisposition has been suggested. If one parent is obese, there are 50% chances for the
children becoming obese. However, no gene alone is responsible for the production of obesity.

Diseases related to Obesity

Sensitivity of peripheral tissues to insulin is decreased. The number of insulin receptors
are decreased in adipose tissue cells. Plasma insulin levels may be elevated. Obesity is associated
with substantially increased cardiovascular risk. Increased waist to hip ratio (abdominal
obesity) is a greater risk. In metabolic syndrome (Syndrome-X) insulin resistance,
hyperglycemia, hyperlipidemia (increased LDL and decreased HDL) and obesity are seen. The
major ill effects of obesity are increased risk of coronary artery disease, diabetes mellitus,
hypertension, metabolic syndrome and a reduced lifespan. Calorie-fat-restricted diet may
retard ageing process and extend the lifespan.
Treatment of Obesity
Lifestyle modification is the best suitable technique. The goal is to reduce the intake of
calories and fat. Frequent small meals with lots of vegetables with controlled exercise is very
useful. In patients with higher grades of obesity, drugs may be given to decrease appetite
(endocannabinoid receptor antagonists will decrease the intake of food).