Pharmaceutical

Biochemistry
(PHS 302)

Dr. Asmaa Saleh Ali

Lipid digestion &
metabolism
Introduction
• The average daily intake of lipids
by adults is about 81 g, of which
more than 90% is normally
triacylglycerol (TAG, formerly
called triglyceride).
• The remainder of the dietary
lipids consists primarily of
cholesterol, cholesteryl esters,
phospholipids, and unesterified
(“free”) fatty acids.
Processing of dietary lipid in the stomach
• The digestion of lipids begins in the stomach, catalyzed by an acid-
stable lipase (lingual lipase) that originates from glands at the back of
the tongue.
• TAG molecules, particularly those containing fatty acids of short- or
medium-chain length (fewer than 12 carbons, such as are found in
milk fat), are the primary target of this enzyme.
• These same TAGs are also degraded by a separate gastric lipase,
secreted by the gastric mucosa. Both enzymes are relatively acid-
stable, with pH optimums of pH 4 to pH 6.
Processing of dietary lipid in the stomach
• These “acid lipases” play a particularly important role in lipid
digestion in neonates, for whom milk fat is the primary source of
calories.
• They also become important digestive enzymes in individuals with
pancreatic insufficiency, such as those with cystic fibrosis.
• Lingual and gastric lipases aid these patients in degrading TAG
molecules (especially those with short- to medium-chain fatty acids)
despite a near or complete absence of pancreatic lipase.
Emulsification of dietary lipid in the small
intestine
• The critical process of emulsification of dietary lipids occurs in the
duodenum.
• Emulsification increases the surface area of the hydrophobic lipid
droplets so that the digestive enzymes, which work at the interface of
the droplet and the surrounding aqueous solution, can act effectively.
• Emulsification is accomplished by two complementary mechanisms,
namely, use of the detergent properties of the bile salts, and
mechanical mixing due to peristalsis.
• Bile salts, made in the liver and stored in the gallbladder, are
derivatives of cholesterol
Degradation of dietary lipids by pancreatic
enzymes
• The dietary TAG, cholesteryl esters, and phospholipids are enzymically
degraded (“digested”) by pancreatic enzymes, whose secretion is
hormonally controlled.
1. TAG degradation:
• TAG molecules are too large to be taken up efficiently by the mucosal
cells of the intestinal villi.
• They are, therefore, acted upon by an esterase, pancreatic lipase,
which preferentially removes the fatty acids at carbons 1 and 3.
• The primary products of hydrolysis are thus a mixture of 2-
monoacylglycerol and free fatty acids
Degradation of dietary lipids by pancreatic
enzymes
2-Cholesteryl ester degradation:
• Most dietary cholesterol is present in the free (nonesterified) form,
with 10–15% present in the esterified form.
• Cholesteryl esters are hydrolyzed by pancreatic cholesteryl ester
hydrolase (cholesterol esterase), which produces cholesterol plus free
fatty acids.
• Cholesteryl ester hydrolase activity is greatly increased in the
presence of bile salts.
Degradation of dietary lipids by pancreatic
enzymes
3. Phospholipid degradation: Pancreatic juice is rich in the proenzyme
of phospholipase A2 that is activated by trypsin and requires bile salts
for optimum activity.
• Phospholipase A2 removes one fatty acid from carbon 2 of a
phospholipid, leaving a lysophospholipid. For example,
phosphatidylcholine becomes lysophosphatidylcholine.
• The remaining fatty acid at carbon 1 can be removed by
lysophospholipase, leaving a glycerylphosphoryl base (for example,
glycerylphosphorylcholine) that may be excreted in the feces, further
degraded, or absorbed.
Control of lipid digestion:
• Pancreatic secretion of the hydrolytic enzymes that degrade dietary
lipids in the small intestine is hormonally controlled. Cells in the
mucosa of the lower duodenum and jejunum produce a small peptide
hormone, cholecystokinin (CCK), in response to the presence of lipids
and partially digested proteins entering these regions of the upper
small intestine. CCK acts on the gallbladder (causing it to contract and
release bile—a mixture of bile salts, phospholipids, and free
cholesterol), and on the exocrine cells of the pancreas (causing them
to release digestive enzymes).
• It also decreases gastric motility, resulting in a slower release of
gastric contents into the small intestine.
Control of lipid digestion:
• Other intestinal cells produce another small
peptide hormone, secretin, in response to the low
pH of the chyme entering the intestine.
• Secretin causes the pancreas and the liver to
release a solution rich in bicarbonate that helps
neutralize the pH of the intestinal contents,
bringing them to the appropriate pH for digestive
activity by pancreatic enzymes.
Absorption of lipids by intestinal
mucosal cells (enterocytes)
• Free fatty acids, free cholesterol, and 2-
monoacylglycerol are the primary products of
lipid digestion in the jejunum.
• These, plus bile salts and fat-soluble vitamins (A,
D, E, and K), form mixed micelles—disk shaped
clusters of amphipathic lipids that coalesce with
their hydrophobic groups on the inside and their
hydrophilic groups on the outside.
• Mixed micelles are, therefore, soluble in the
aqueous environment of the intestinal lumen
Absorption of lipids by intestinal mucosal
cells (enterocytes)
• The hydrophilic surface of the micelles facilitates the transport of the
hydrophobic lipids to the brush border membrane where they are
absorbed. Bile salts are absorbed in the ileum. [Note: Relative to
other dietary lipids, cholesterol is only poorly absorbed by the
enterocytes.
• Drug therapy (for example, ezetimibe) can further reduce cholesterol
absorption in the small intestine. Short- and medium chain length
fatty acids do not require the assistance of mixed micelles for
absorption by the intestinal mucosa.
Resynthesis of TAG and cholesteryl esters
• The mixture of lipids absorbed by the enterocytes migrates to the endoplasmic
reticulum. Fatty acids are and 2-monoacylglycerols absorbed by the enterocytes are
converted to TAGs by the enzyme complex, TAG synthase.
• Cholesterol is esterified to a fatty acid primarily by acyl CoA: cholesterol
acyltransferase.
• Virtually all long-chain fatty acids entering the enterocytes are used to form TAGs,
phospholipids, and cholesteryl esters.
• Short- and medium-chain length fatty acids are not converted to their CoA
derivatives, and are not reesterified to 2-monoacylglycerol. Instead, they are released
into the portal circulation, where they are carried by serum albumin to the liver.
Lipid malabsorption
• Lipid malabsorption, resulting in increased lipid
(including the fat soluble vitamins and essential fatty
acids) in the feces (steatorrhea), can be caused by
disturbances in lipid digestion and/or absorption. Such
disturbances can result from several conditions,
including Cystic fibrosis (causing poor digestion) and
shortened bowel (causing decreased absorption).
• The ability of short- and medium-chain length fatty
acids to be taken up by enterocytes without the aid of
mixed micelles has made them important in dietary
therapy for individuals with malabsorption disorders.
Secretion of lipids from enterocytes
• The newly resynthesized TAGs and cholesteryl esters are very
hydrophobic, and aggregate in an aqueous environment.
• It is, therefore, necessary that they be packaged as particles of lipid
droplets surrounded by a thin layer composed of phospholipids,
unesterified cholesterol, and a molecule of the characteristic protein,
apolipoprotein B-48.
• This layer stabilizes the particle and increases its solubility, thereby
preventing multiple particles from coalescing.
• The particles are named chylomicrons
 Fatty Acid and
Triacylglycerol Metabolism
Fatty Acid and Triacylglycerol Metabolism
• Fatty acids exist “free” in the body (that is, they are unesterified), and
are also found as fatty acyl esters in more complex molecules, such as
triacylglycerols.
• Free fatty acids can be oxidized by many tissues —particularly liver
and muscle—to provide energy.
• Esterified fatty acids, in the form of triacylglycerols stored in adipose
cells, serve as the major energy reserve of the body
lipogenesis
De novo synthesis of fatty acids
• large proportion of the fatty acids used by the body is supplied by the diet.
Carbohydrates and protein obtained from the diet in excess of the body’s
needs for these compounds can be converted to fatty acids, which are
stored as triacylglycerols.  
• In adult humans, fatty acid synthesis occurs primarily in the liver and
lactating mammary glands and, to a lesser extent, in adipose tissue.
• This cytosolic process incorporates carbons from acetyl coenzyme A (CoA)
into the growing fatty acid chain, using adenosine triphosphate (ATP) and
reduced nicotinamide adenine dinucleotide phosphate (NADPH).
A. Production of cytosolic
acetyl CoA
• The first step in de novo fatty acid synthesis is the
transfer of acetate units from mitochondrial acetyl
CoA to the cytosol.
• The CoA portion of acetyl CoA, however, cannot
cross the inner mitochondrial membrane; only the
acetyl portion enters the cytosol.
• It does so as part of citrate produced by the
condensation of oxaloacetate (OAA) and acetyl CoA.
• This process occurs when the mitochondrial citrate
concentration is high.
B. Carboxylation of acetyl CoA
 to form malonyl CoA
• The carboxylation of acetyl CoA to form
malonyl CoA is catalyzed by acetyl CoA
carboxylase , and requires CO2 and ATP.
• The coenzyme is the vitamin, biotin
C. Fatty acid synthase: a multifunctional
enzyme in eukaryotes
• The remaining series of reactions of fatty acid synthesis in eukaryotes
is catalyzed by the multifunctional, dimeric enzyme, fatty acid
synthase (FAS).
Further elongation of fatty acid chains
• Although palmitate, a 16-carbon, fully saturated long-chain length
fatty acid (16:0), is the primary end product of fatty acid synthase
activity, it can be further elongated by the addition of two-carbon
units in the smooth endoplasmic reticulum (SER).
• Elongation requires a system of separate enzymes rather than a
multifunctional enzyme. Malonyl CoA is the two-carbon donor and
NADPH supplies the electrons.
• The brain has additional elongation capabilities, allowing it to produce
the very-long-chain fatty acids (over 22 carbons) that are required for
synthesis of brain lipids.
Desaturation of fatty acid chains
• Enzymes (desaturases) also present in the SER are responsible for
desaturating long-chain fatty acids (that is, adding cis double bonds).
The desaturation reactions require NADH, cytochrome b5 and its FAD-
linked reductase.
Storage of fatty acids as components of
triacylglycerols
• Mono-, di-, and triacylglycerols consist of one, two, or three
molecules of fatty acid esterified to a molecule of glycerol.
• Because TAGs are only slightly soluble in water and cannot form
stable micelles by themselves, they coalesce within adipocytes to
form oily droplets that are nearly anhydrous. These cytosolic lipid
droplets are the major energy reserve of the body.
Synthesis of triacylglycerol
• A. Synthesis of glycerol phosphate:
• There are two pathways for glycerol phosphate production.
• In both liver (the primary site of TAG synthesis) and adipose tissue,
glycerol phosphate can be produced from glucose, using first the
reactions of the glycolytic pathway to produce dihydroxyacetone
phosphate. Next, DHAP is reduced by glycerol phosphate
dehydrogenase to glycerol phosphate.
• A second pathway found in the liver, but not in adipose tissue, uses
glycerol kinase to convert free glycerol to glycerol phosphate
Synthesis of triacylglycerol
Synthesis of triacylglycerol
• B. Conversion of a free fatty acid to its activated form:
• A fatty acid must be converted to its activated form (attached to CoA)
before it can participate in metabolic processes such as TAG synthesis.
• This reaction is catalyzed by a family of fatty acyl CoA synthetases
(thiokinases).
Synthesis of triacylglycerol
C. Synthesis of a molecule of
TAG from glycerol phosphate
and fatty acyl CoA:
• This pathway involves four
reactions.
• These include the sequential
addition of two fatty acids
from fatty acyl CoA, the
removal of phosphate, and
the addition of the third fatty
acid.
Mobilization of stored fats and
oxidation
of fatty acids
(lipolysis)
• Fatty acids stored in adipose tissue, in the form of neutral TAG, serve
as the body’s major fuel storage reserve.
• TAGs provide concentrated stores of metabolic energy because they
are highly reduced and largely anhydrous.
• The yield from the complete oxidation of fatty acids to CO2 and H2O
is 9 kcal/g fat (as compared to 4 kcal/g protein or carbohydrate)
Release of fatty acids from TAG

• The mobilization of stored fat requires the hydrolytic release of fatty

acids and glycerol from their TAG form.
• This process is initiated by hormone-sensitive lipase, which removes a
fatty acid from carbon 1 and/or carbon 3 of the TAG.
• Additional lipases specific for diacylglycerol or monoacylglycerol
remove the remaining fatty acid(s).
Release of fatty acids from TAG
Activation of hormone-sensitive
lipase (HSL):
• This enzyme is activated when phosphorylated by a
3 ',5 '-cyclic AMP(cAMP)– dependent protein kinase.
3',5'-Cyclic AMP is produced in the adipocyte when
one of several hormones (such as epinephrine or
glucagon) binds to receptors on the cell membrane,
and activates adenylyl cyclase.
• In the presence of high plasma levels of insulin and
glucose, HSL is dephosphorylated, and becomes
inactive.
Fate of glycerol:
• The glycerol released during TAG degradation cannot be metabolized
by adipocytes because they apparently lack glycerol kinase.
• Rather, glycerol is transported through the blood to the liver, where it
can be phosphorylated.
• The resulting glycerol phosphate can be used to form TAG in the liver,
or can be converted to DHAP by reversal of the glycerol phosphate
dehydrogenase reaction. DHAP can participate in glycolysis or
gluconeogenesis.
Fate of fatty acids:
• The free (unesterified) fatty acids move through the cell membrane of
the adipocyte, and bind to plasma albumin.
• They are transported to the tissues, enter cells, get activated to their
CoA derivatives, and are oxidized for energy.
• Regardless of their levels, plasma free fatty acids (FFA) cannot be used
for fuel by erythrocytes, which have no mitochondria.
• Brain, too, does not use fatty acids for energy, but the reasons are less
clear
β-Oxidation of fatty acids
• The major pathway for catabolism of fatty acids is a mitochondrial
pathway called β-oxidation, in which two-carbon fragments are
successively removed from the carboxyl end of the fatty acyl CoA,
producing acetyl CoA, NADH, and FADH2.
Transport of long-chain fatty acids (LCFA)
into the mitochondria:
• After a LCFA enters a cell, it is converted in the cytosol to its CoA
derivative by long-chain fatty acyl CoA synthetase (thiokinase), an
enzyme of the outer mitochondrial membrane.
• Because β-oxidation occurs in the mitochondrial matrix, the fatty acid
must be transported across the inner mitochondrial membrane that is
impermeable to CoA. Therefore, a specialized carrier transports the
long-chain acyl group from the cytosol into the mitochondrial matrix.
• This carrier is carnitine, and this rate-limiting transport process is
called the carnitine shuttle
Entry of short- and medium-chain fatty
acids into the mitochondria:
• Fatty acids shorter than 12 carbons can cross the inner mitochondrial
membrane without the aid of carnitine system.
• Once inside the mitochondria, they are activated to their CoA
derivatives by matrix enzymes, and are oxidized.
• Medium-chain fatty acids are plentiful in human milk.
Reactions of β-oxidation:
• The first cycle of β-oxidation is shown in Figure 16.17.
• It consists of a sequence of four reactions involving the
β-carbon (carbon 3) that results in shortening the fatty
acid chain by two carbons.
• The steps include an oxidation that produces FADH2, a
hydration step, a second oxidation that produces NADH,
and a thiolytic cleavage that releases a molecule of
acetyl CoA.
• These four steps are repeated for saturated fatty acids of
even-numbered carbon chains (n/2) – 1 times (where n
is the number of carbons), each cycle producing an
acetyl group plus one NADH and one FADH2.
• The final thiolytic cleavage produces two acetyl groups.
Energy yield from fatty acid oxidation:
• The energy yield from the -oxidation pathway is high. For example,
the oxidation of a molecule of palmitoyl CoA to CO2 and H2O
produces 8 acetyl CoA, 7 NADH, and 7 FADH2, from which 131 ATP
can be generated; however, activation of the fatty acid requires 2 ATP.
Thus, the net yield from palmitate is 129 ATP
Calculation of energy production of oxidation of any fatty acid:
• = {(N/2 – 1) × 5 ATP} + {N/2 x 12 ATP} – 2 ATP
• Where N = Number of carbons of fatty acid e.g. palmitic acid = 16 carbons, so
energy production =
• ={(16/2 – 1) × 5 ATP} + {16/2 x 12 ATP} – 2 ATP
• ={(8 – 1) × 5 ATP} + {8 x 12 ATP} – 2 ATP = 129 A
e.g. palmitic acid (16 carbons):
• -oxidation of palmitic acid will be repeated 7 times (turns) to
produce 8 acetyl CoA.
• In each turn, one molecule of reduced FADH2 and one molecule of
reduced NADH+H are produced. They are oxidized in respiratory
chain to give 5 ATP.
• FADH2  2 ATP
• NADH+H  3 ATP
• 7 turns  5 ATP  35 ATP.
• Oxidation of one molecule of acetyl CoA in citric acid cycle gives 12
ATP.
• 8 Acetyl CoA  12 ATP = 96 ATP
• Two high energy phosphate bonds are utilized in the first reaction
(catalyzed by acyl CoA synthetase) which occurs for one time only.
• - 2 ATP
•  Net energy gain = (35 ATP + 96 ATP) – 2 ATP = 129 ATP 55
α-Oxidation of fatty acids
• Branched-chain, 20 carbon fatty acid,
phytanic acid:
• This is not a substrate for acyl CoA
dehydrogenase because of the methyl
group on its β carbon.
• Instead, it is hydroxylated at the α-carbon
by phytanoyl CoA α-hydroxylase (PhyH),
carbon 1 is released as CO2, and the
product, 19 carbon pristanic acid, is
activated to its CoA derivative and
undergoes β-oxidation.
α-Oxidation of fatty acids
• Refsum disease is a rare, autosomal
recessive disorder caused by a deficiency
of peroxisomal PhyH.
• This results in the accumulation of
phytanic acid in the plasma and tissues.
• The symptoms are primarily neurologic,
and the treatment involves dietary
restriction to halt disease progression.
 - Oxidation of fatty acids :
• It is oxidation of terminal CH3
group of fatty acid.
• It produces dicarboxylic fatty acids.
• By  oxidation they are converted
to adipic acid (6 carbons) and
suberic acid (8 carbons).
• It is a minor pathway for fatty acid
oxidation and catalyzed by
hydroxylase enzymes of
cytochrome P450. 59