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
 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 Phospholipase A1
 The site of action of different
O
phospholipids is shown
Phospholipase A2 CH2 O C CH2 R1
below. 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
2H O Free fatty acid Free fatty acid
CH2 O C CH2 R3 2 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
 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
Lymph Enterocyte
Enterocyte Intestinal
Intestinal
Lumen
Lumen

2 Fatty Acid

I
+
Monoglycerid

T
e

DGAT

Triglyceride
 Phospholipid Absorption

Intestinal
Intestinal
Lumen
Lumen
Lymph
Lymph Enterocyte
Enterocyte

Fatty Acid

I
+
Lysophospholipid

Phospholipid
 Chylomicron Formation

Intestinal
Intestinal
Lymph
Lymph Enterocyte
Enterocyte Lumen
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 genetic defects of lipid absorption (e.g.,
 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 non-esterified fatty acids.

 HSL is activated by epinephrine, nor-epinephrine, ACTH

and glucagon, acting via phosphorylation of the enzyme.

Adipocytes release nonesterified fatty acids
 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: PP i + 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 inter-membrane 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.
 Transport
 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 carnitine palmitoyl-carnitine

Space
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 + CoAAMP + 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
 Palmitoylcarnitine

inner mitochondrial Carnitine

membrane respiratory chain
translocase

Palmitoylcarniti
matrix side ne 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
 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.
 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 (LCAD) acts on
chains greater than C12.
 Medium chain fatty acyl CoA dehydrogenase (MCAD)
acts on chains of C6 to C12.
 Short chain fatty acyl CoA dehydrogenase (SCAD) acts
on chains of C4 to C6.
 MCAD deficiency is thought to be one of the most
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
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.