Digestion and absorption of lipids

Major dietary lipids are

Triacylglycerol – 90-95 %
Cholesterol ester/Cholestrol
Phoshpolipids
 Digestion of lipids

Digestion in mouth
There is hardly any digestion of fats in the mouth-
even though lingual lipase is available.
 Is secreted from Ebner's glands of the tongue
Its pH optima is on the acidic side, therefore, its
activity is mainly in the stomach
 Digestion in the stomach

Lingual lipase
Works mainly in the stomach, particularly within the
core of food bolus
 to hydrolyze short-chain fatty acids at C1/C3 of glycerol.
Therefore, lingual lipase is able to digest the fats in cow's
milk, butter, ghee and coconut oil.
The released water-soluble fatty acids are absorbed from
stomach wall to the portal circulation.
Lingual lipase may hydrolyze 30% of dietary
triacylglycerols.
 Gastric lipase
 Gastric lipase is secreted by the chief cells in response to gastrin.
 It has a wide pH range near neutrality and requires calcium for
activation.
 It cannot work without emulsification of fat and within the acidic
pH of the stomach.
 Gastric lipase has an important role in the milk fat digestion in the
stomach of infants since the pH of stomach in infants is not as
acidic as in adults.
 The presence of fat in stomach stimulates secretion of
enterogasterone hormone, which delays the gastric emptying time
of food.
 The longer stay of food in the stomach gives the characteristic
high satiety value.
 Digestion in small Intestine
 Digestion of lipids begins in the duodenum, when the entrance of the
acid chyme from the stomach stimulates the secretion of enteric
hormones (small peptides) by the duodenal mucosa.

 Gastric HCl stimulates the secretion of secretin that promotes the

release of HCO3--rich juice from the pancreas into the duodenum via
the pancreatic duct.

 Presence of fat in small intestine stimulate the release of

cholecystokinin from intestinal mucosa to the blood to induce the
release of zymogen granules from the pancreas and the contraction of
the gall bladder to release of bile.
 Pancreatic enzymes
The pH of the intestine rises to alkaline side due to
the presence of bicarbonate in the pancreatic
juice. Three lipid-specific proenzymes are
activated by the bile acids and the neutral pH.
These enzymes are:
Phospholipase A2
Cholesterol esterase
Pancreatic lipase, also known as Steapsin
Phospholipase A2 is secreted as proenzyme from the
pancreas that is activated by trypsin.
It hydrolyzes fatty acid at C2 of glycerophospholipids.
Other intestinal phospholipases and phosphatases
complete the hydrolysis of phospholipids into glycerol,
free fatty acids, choline and phosphate. But the main
product is the lysophospholipids.
Phospholipids + H2O  Lysophospholipids + free fatty acids
 The site of action of different phospholipids is shown below.
Phospholipase A1
O
Phospholipase A2 CH2 O C CH2 R1
O
R2 CH2 C O CH CH3
O
CH2 O P O CH2 CH2 N+ CH3
OH CH3
Phospholipase C Phospholipase D
Digestion of phospholipids
Cholesterol esterase from the pancreas reversibly
hydrolyzes cholesterol esters. It has an optimum pH of
8.0.
Cholesterol esters + H2O  Cholesterol + free fatty
acid
Pancreatic lipase, also known as Steapsin, cleaves triacylglycerols to
2-monoacylglycerol and two free fatty acids.
It has an optimum pH of 6.0 that is furnished by the cosecreted
pancreatic bicarbonate.

O
CH2 O C CH2 R1 CH2 OH
O O O O
Pancreatic lipase
R2 CH2 C O CH R2 CH2 C O CH + HO C CH2 R1 + HO C CH2 R3
O
2H2O Free fatty acid Free fatty acid
CH2 O C CH2 R3 CH2 OH
A triglyceride (Triacylglycerol) 2-monoacylglycerol
 Absorption of lipids
End products of fat digestion are
Lysophospholipids
Monoacylglycerols
FFAs
Free cholesterol
Glycerol
 Are solubilized in the intestine by bile acid to form
micelles that fuses with plasma membrane of the
intestinal villar mucosal cells of the jejunum and ileum
to shuttle the content into intestinal cells
 Fats
Dietary fats constitute 20 - 40 % of the daily calories with
triglycerides as the major portion. However, fats are
recommended not to exceed 30% of the daily caloric
requirement.
 Only 22% of triacylglycerols are completely hydrolyzed into
glycerol and FFAs.
 Most of the exogenous triglycerides are absorbed as 2-
monoacylglycerol (72%) because of inability of pancreatic
lipase to hydrolyze fatty acid at C2 of glycerol.
 The 1-monoacylglycerol produced by isomerization of fatty
acid from C2 to C1 may be hydrolyzed by pancreatic lipase or
absorbed as it is (6%).
 Inside the mucosal cells 1-monoacylglycerols are hydrolyzed
into glycerol and free fatty acids by intestinal lipase
 Re-esterification
keeps a sharp gradient of concentration within
the mucosal cells that favors the continuous rapid
diffusion of digested lipids into the blood.
A thiokinase (acyl-CoA synthetase) activates fatty
acids into fatty acyl coenzyme A (acyl-CoAs).
Glycerol is activated by glycerokinase to glycerol-
3-phosphate that combines with three acyl-CoAs
to regenerate triacylglycerols.
Cholesterol and lysophospholipids are re-
esterified in a similar manner into cholesterol
esters and phospholipids.
 Triglyceride Absorption

Lymph Enterocyte Intestinal

Lumen

2 Fatty Acid

I
+
Monoglycerid

T
e

DGAT

Triglyceride
 Phospholipid Absorption

Intestinal
Lumen
Lymph Enterocyte

Fatty Acid

I
+
Lysophospholipid

Phospholipid
 Chylomicron Formation

Intestinal
Lymph Enterocyte Lumen

Phospholipid

Triglyceride

With
apoB48 Cholesteryl
Ester
 Steatorrhea

The presence of fat in feces, at an amount more than normal (<5%), is known as
steatorrhoea.
The condition is due to improper fat digestion and/or absorption. Possible
causes are:
Pancreatic lipase deficiency or deficiency of the colipase: It is due to acute
or chronic pancreatitis (e.g., alcohol-induced), pancreatic duct
obstruction, cystic fibrosis.
It causes defective digestion of fat that also hinders digestion of other
foodstuffs. Stools contain undigested fat, protein and other food
constituents that are coated by the fat and prevented from digestion.
Most of fat-soluble vitamins are absorbed.
Deficiency of bile salts, phospholipids or calcium: It is due to liver diseases
that prevent synthesis and/or secretion of bile acids, and obstruction of
bile duct (due to stones or tumors.
Failure of intestinal digestion and absorption: Due to the damaged
intestinal mucosal cells in certain diseases such as Crohn's diseases, and
 LIPID METABOLISM
General considerations
Lipolysis, oxidation of fatty acids
B-oxidation of fatty acids
 General Overview
A. Fat metabolism
Fat metabolism in adipose tissue
Fats (triacylglycerols) are the most important
energy reserve in the animal organism.
They are mostly stored in insoluble form in the
cells of adipose tissue,
The adipocytes: where they are constantly being
synthesized and broken down again.
 Biosynthesis of fats
 As precursors for the biosynthesis of fats (lipogenesis), the
adipocytes use triacylglycerols from lipoproteins (VLDLs and
chylomicrons;), which are formed in the liver and intestines
and delivered by the blood.

 Lipoprotein lipase , which is located on the inner surface of

the blood capillaries, cleaves these triacylglycerols into
glycerol and fatty acids, which are taken up by the adipocytes
and converted back into fats.
 Degradation of fats (Lipolysis)
 The degradation of fats (lipolysis) is catalyzed in adipocytes by
hormone-sensitive lipase
 It is an enzyme that is regulated by various hormones by cAMP-
dependent interconversion.
 The amount of fatty acids released depends on the activity of this
lipase; in this way, the enzyme regulates the plasma levels of fatty
acids.
 In the blood plasma, fatty acids are transported in free form i. e.,
non-esterified.
 Only short-chain fatty acids are soluble in the blood; longer, less
water-soluble fatty acids are transported bound to albumin.
 Degradation of fatty acids in the liver
The metabolism of fatty acids is particularly intensive in the
hepatocytes in the liver.
 The most important process in the degradation of fatty acids is ß-
oxidation in the mitochondrial matrix.
 Initially, the fatty acids in the cytoplasm are activated by binding to
coenzyme A into acyl CoA.
 Then, with the help of a transport system (the carnitine shuttle, the
activated fatty acids enter the mitochondrial matrix, where they are
broken down into acetyl CoA.
 The resulting acetyl residues can be oxidized to CO2 in the
tricarboxylic acid cycle, producing reduced coenzyme and ATP
derived from it by oxidative phosphorylation.
Fat synthesis in the liver
 Fatty acids and fats are mainly synthesized in the liver
and in adipose tissue, as well as in the kidneys, lungs,
and mammary glands.
 Fatty acid biosynthesis occurs in the cytoplasm in
contrast to fatty acid degradation.
 The most important precursor is glucose, but certain
amino acids can also be used.
 Fat synthesis….
 The first step is carboxylation of acetyl CoA to malonyl
CoA. This reaction is catalyzed by acetyl-CoA
carboxylase, which is the key enzyme in fatty acid
biosynthesis.
 Synthesis into fatty acids is carried out by fatty acid
synthase.
 This multifunctional enzyme starts with one molecule of
acetyl- CoA and elongates it by adding malonyl groups in
seven reaction cycles until palmitate is reached.
 One CO2 molecule is released in each reaction cycle and
fatty acids will grows by two carbon units each time.
Fat synthesis…
NADPH+H+ is used as the reducing agent and is derived
either from the PPP or from isocitrate DH and malic
enzyme reactions.
The elongation of the fatty acid by fatty acid synthase
concludes at C16, and the product, palmitate (16:0), is
released.
Unsaturated fatty acids and long-chain fatty acids can arise
from palmitate in subsequent reactions.
Fats are finally synthesized from activated fatty acids (acyl
CoA) and glycerol 3-phosphate.
To supply peripheral tissues, fats are packed by the
hepatocytes into lipoprotein complexes of the VLDL type
and released into the blood in this form.
 Mobilization and Transport of Adipose
Fatty Acid
Fatty acids, as triacylglycerol (triglyceride), are
stored in white adipose tissue.
This fat storage comprises the 100,000 Kcal of
energy stored as fat.
Brown adipose tissue is a thermogenic tissue,
and is not important in energy storage.
Storage in adipocytes as triacylglycerol
Fatty acids are stored primarily in adipocytes as
triacylglycerol.
Triacylglycerol must be hydrolyzed to release the fatty
acids.
Adipocytes are found mostly in the abdominal cavity
and subcutaneous tissue.
Adipocytes are metabolically very active; their stored
triacylglycerol is constantly hydrolyzed and
resynthesized.
Mobilization of Stored Fats: Lipolysis
Adipocytes release nonesterified fatty acids
 Non-esterified fatty acid release from the adipocytes is initiated by
the action of hormone sensitive lipase (HSL), which begins to
hydrolyze the stored triglyceride.
 The final products of triacylglycerol hydrolysis are glycerol and
nonesterified fatty acids.
 HSL is activated by epinephrine, norepinephrine, ACTH and
glucagon, acting via phosphorylation of the enzyme.
 It is inhibited by insulin.
 Non-esterified fatty acids are bound to serum albumin for
transport to other tissues, where they are used.
 Major target tissues are muscle and liver.
 At the target cells non-esterified fatty acids are taken up passively.
Within the target cells they are bound to fatty acid binding
protein. Next they must be activated.
 Fatty Acid Activation and Transport into the
Mitochondria
Fatty acids inside the cell, like glucose, must be
activated before proceeding through
metabolism.
Activation consists of conversion of the non-
esterified fatty acid to its CoA derivative.
The activated fatty acid may then be transported
into the mitochondrion for energy production.
 Activation
Fatty acids are activated by fatty acyl CoA synthetase.
The reaction:
R-COOH + CoASH + ATP <--> R-CO-SCoA + AMP + PPi
 The subsequent hydrolysis of PPi draws the reaction in the
forward direction, maintaining a low cytosolic free fatty
acid concentration: PPi + H2O --> 2 Pi
 The reaction occurs in the endoplasmic reticulum and the
outer mitochondrial membrane.
 Transport
 The fatty acyl group is transported into the mitochondrial
matrix, where it undergoes beta-oxidation.
 In the intermembrane space of the mitochondria, fatty acyl
CoA reacts with carnitine in a reaction catalyzed by
carnitine acyltransferase I (CAT-I), yielding CoA and fatty
acyl carnitine. The resulting fatty acyl carnitine crosses the
inner mitochondrial membrane.
 CAT-I is associated with the inner leaflet of the outer
mitochondrial membrane.
 In liver the CAT-I reaction is rate-limiting; the enzyme is
allosterically inhibited by malonyl CoA. Malonyl CoA
concentration would be high during fatty acid synthesis.
 Inhibition of CAT-I by malonyl CoA prevents simultaneous
synthesis and degradation of fatty acids.
 Transport and Regeneration
 Fatty acyl CoA is impermeable to the inner
mitochondrial membrane, so it is carried in the form of
fatty acyl carnitine.
 Fatty acyl carnitine is transported across the inner
mitochondrial membrane in exchange for carnitine by an
antiport translocase.
 In the mitochondrial matrix fatty acyl carnitine reacts
with CoA in a reaction catalyzed by carnitine
acyltransferase II (CAT-II), yielding fatty acyl CoA and
carnitine.
 The fatty acyl CoA is now ready to undergo beta-
oxidation.
 ATP + CoA AMP + PPi

palmitate palmitoyl-CoA

Cytoplasm

OUTER
ACS MITOCHONDRIAL
CPT-I
[1] [2] MEMBRANE

CoA
palmitoyl-CoA

Intermembrane palmitoyl-carnitine
carnitine
Space
CPT-I defects cause severe muscle weakness because fatty acids are an
important muscle fuel during exercise.

Activation of palmitate to palmitoyl CoA and conversion to palmitoyl carnitine

 CPT-I

palmitoyl-CoA CoA

Intermembrane Space carnitine palmitoyl-carnitine

INNER
CAT [3] MITOCHONDRIAL
MEMBRANE

Matrix CPT-II
carnitine palmitoyl-carnitine
[4]
palmitoyl-CoA CoA

Mitochondrial uptake via of palmitoyl-carnitine via the carnitine-

acylcarnitine translocase (CAT).
 ATP + CoA AMP + PP
i
Cytoplasm
palmitate palmitoyl-CoA

OUTER
ACS MITOCHONDRIAL
CPT-I
MEMBRANE
[1] [2]

CoA Intermembrane
palmitoyl-CoA Space

carnitine palmitoyl-carnitine

INNER
[3] MITOCHONDRIAL
CAT MEMBRANE

CPT-II
carnitine palmitoyl-carnitine
Matrix
[4]
palmitoyl-CoA CoA
 Mitochondrial beta-oxidation
 Beta-oxidation is the process by which long chain fatty acyl CoA is
degraded. The products of beta-oxidation are:
 acetyl CoA
 FADH2, NADH and H+
The overall reaction, using palmitoyl CoA (16:0) as a model substrate:
7 FAD + 7 NAD+ + 7 CoASH + 7 H2O + H(CH2CH2)7CH2CO-SCoA --> 8
CH3CO-SCoA + 7 FADH2 + 7 NADH + 7 H+

Fate of acetyl CoA

 Oxidation by the citric acid cycle.
 In liver only, acetyl CoA may be used for ketone body synthesis.
Fate of the FADH2 and NADH + H+
FADH2 and NADH + H+ are oxidized by the mitochondrial electron
transport system, yielding ATP.
 Palmitoylcarnitine

inner mitochondrial Carnitine

membrane respiratory chain
translocase

Palmitoylcarnitine
matrix side 2 ATP
3 ATP
Palmitoyl-CoA
FAD
oxidation
FADH2
hydration H2O

recycle NAD+
oxidation
6 times
NADH
thiolase CoA

CH3-(CH)12-C-S-CoA + Acetyl CoA

Citric
O acid
cycle 2 CO2
Four enzymes and reactions
There are four individual reactions of beta-oxidation,
each catalyzed by a separate enzyme.
Dehydrogenation between the alpha and beta carbons
(C2 and C3) in a FAD-linked reaction.
Hydration of the double bond by enoyl CoA hydratase.
A second dehydrogenation in a NAD-linked reaction.
Thiolytic cleavage of the thioester by beta-ketoacyl CoA
thiolase.
This sequence of four steps is repeated until the fatty
acyl chain is completely degraded to acetyl CoA.
 Dehydrogenation
 There are three fatty acyl CoA dehydrogenases. Each is specific for
a different acyl chain length, so different enzymes are involved in
different stages of beta-oxidation.
 Long chain fatty acyl CoA dehydrogenase acts on chains greater >
C12.
 Medium chain fatty acyl CoA dehydrogenase acts on chains of C6 to
C12.
 Short chain fatty acyl CoA dehydrogenase acts on chains of C4 to C6.

 MCAD deficiency is thought to be one of the most common inborn

errors of metabolism
 Dehydrogenation
Dehydrogenation occurs between the alpha and beta carbons (C2 and C3) in
a FAD-linked reaction catalyzed by acyl CoA dehydrogenase.
The product contains a trans- double bond. Involvement of the beta-carbon
in this and subsequent steps gives the pathway its name.

Hydration
Hydration of the double bond is catalyzed by enoyl CoA
hydratase. The product is an L-3-hydroxyacyl CoA.
2nd dehydrogenation
A second dehydrogenation, of the alcohol, occurs
in a NAD-linked reaction catalyzed by beta-
hydroxyacyl CoA dehydrogenase. The product is
a ketone.
 Thiolytic cleavage
 Thiolytic cleavage of the thioester is catalyzed by beta-ketoacyl CoA thiolase.
 Reaction products: The products are acetyl CoA and a long chain fatty acyl CoA
that is two carbons shorter than the original fatty acyl CoA.

 The shortened fatty acyl group is now ready for another round of beta-oxidation.
After the fatty acyl CoA has been reduced to acetyl or propionyl CoA, beta-
oxidation is complete.
 Regulation: This reaction is inhibited by high concentrations of acetyl CoA.
 Beta-oxidation is regulated as a whole primarily by fatty acid availability; once
fatty acids are in the mitochondria they are oxidized as long as there is adequate
NAD+ and CoA.
Complete beta-oxidation of palmitoyl CoA

Complete beta-oxidation of palmitoyl CoA:

7 FAD + 7 NAD+ + 7 CoASH + 7 H2O + H(CH2CH2)7CH2CO-

SCoA --> 8 CH3CO-SCoA + 7 FADH2 + 7 NADH + 7 H+
ENERGY YIELD FROM ß-OXIDATION
From PalmitoylCoA ATP Yield
7NADH x 3 ATP by ETC oxidation 21
7 FADH2 x 2 ATP by ETC oxidation 14
8 Acetyl CoA x 12 ATP via Krebs CAC 96

Total (Gross) 131 ATP

Less 2 ATP
NET 129 ATP
From one molecule of palmitoylCoA
F.A Synthesis
The rest covered by your self Like
unsaturated fatty acids and odd
chain fatty acids metabolism
Thank you
Additional Enzymes
Additional enzymes are needed for complete
oxidation of unsaturated and odd-carbon fatty
acids.
The action of enoyl CoA isomerase may be
required.
A system is needed to generate the trans- double
bond required in beta-oxidation in place of the cis-
bond which occurs naturally in fatty acids.
The three-carbon propionyl CoA residue from
beta-oxidation of odd-chain fatty acids is
metabolized with special enzymes.
Additional Enzymes: Enoyl CoA
The action of enoyl CoA isomerase is required to handle
double bonds at odd-numbered carbons because beta-
oxidation generates or requires pre-existing double bonds
at even-numbered carbons.
If there is a double bond at an odd-numbered carbon (e.g.,
18:1 9), the action of enoyl CoA isomerase is required to
move the naturally occurring cis- bond and convert it to
the trans- bond used in beta-oxidation
The product, with a trans- double bond, is a substrate for
enoyl CoA hydratase, the second enzyme of beta-
oxidation.
Additional Enzymes: trans- vs cis
 Generating a trans- instead of a cis- double bond.
 If there is also a double bond at an even-numbered carbon (e.g.,
the second double bond in 18:2 9,12), the problem is to generate a
trans- double bond instead of a cis-. This occurs in an indirect
manner. Both activities occur in the mitochondrial matrix.
 FIRST: Three cycles of beta-oxidation occur normally. Beta-
oxidation then continues as expected in the presence of the 9
double bond through the fourth cycle, generating a trans-
double bond at the 2-position.
 The fourth cycle completes, and the fifth cycle then begins
normally, but proceeds only through the acyl CoA
dehydrogenase step.
SECOND: 2,4-dienoyl CoA reductase reduces the
compound, leaving one trans- double bond, but in
the wrong position. NADPH + H+ is required.
The product is a substrate for enoyl isomerase, the
same enzyme used for cis- double bonds at odd-
numbered carbons. It moves the double bond from
the 3 to the 2 position.

Beta-oxidation now proceeds normally

Additional Enzymes: Propionyl CoA

Handling the three-carbon propionyl CoA

Fatty acids with an odd number of carbons in their
chains require a means of handling the three-carbon
propionyl CoA that is the final fragment produced by
beta-oxidation of such a chain:
The first step is carboxylation by the biotin-
dependent propionyl CoA carboxylase in an ATP-
requiring reaction.

The D- isomer, which is the product, is then

converted to the L- isomer by methylmalonyl CoA
racemase.
In the final step, the L- isomer is converted to
succinyl CoA by methylmalonyl CoA mutase

Succinyl CoA can then be metabolized through

the tricarboxylic acid cycle.
Summary
Complete Oxidation of an Odd-Chain Fatty Acid --
Summary

This diagram shows production of propionyl CoA from

an odd-chain fatty acid and the subsequent conversion
of propionyl CoA to succinyl CoA, which can be
metabolized through the citric (tricarboxylic) acid cycle.
 Synthesis and Utilization of Ketone Bodies

Overview

These are the compounds known as ketone bodies.

Notice that beta-hydroxybutyrate is not chemically a
ketone.
It is considered to be physiologically equivalent to one
because it and acetoacetate are readily interconverted in
the body.

Synthesis from acetyl CoA
Ketone bodies are synthesized from acetyl CoA.
 Ketone body synthesis from acetyl CoA occurs in hepatic
mitochondria.
 First, acetoacetate is produced in a three-step process.
 Acetoacetate can be reduced to beta-hydroxybutyrate.
 Acetone also arises in small amounts as a biologically inert
side product.
 Ketone body production is regulated primarily by
availability of acetyl CoA.
 If mobilization of fatty acids from adipose tissue is high,
hepatic beta-oxidation will occur at a high rate, and so will
synthesis of ketone bodies from the resulting acetyl CoA.
 The rate of ketone body production increases in starvation.
Synthesis from acetyl CoA: Step 1
The first step is formation of acetoacetyl CoA in a
reversal of the thiolase step of beta-oxidation.
 Synthesis from acetyl CoA: Step 2
In the second step, a third molecule of acetyl CoA
condenses with the acetoacetyl CoA, forming 3-hydroxy-3-
methylglutaryl CoA (HMG CoA) in a reaction catalyzed by
HMG CoA synthase.
 Synthesis from acetyl CoA: Step 3
In the third step HMG CoA is cleaved to yield
acetoacetate (a ketone body) in a reaction catalyzed by
HMG CoA lyase (HMG CoA cleavage enzyme). One
molecule of acetyl CoA is also produced.