FA oxidation and ketone bodies

Roles of Lipids

• principal form of stored energy

• major constituents of cell membranes
• vitamins
• messengers – intra and extracellular
 Starved state
Glycerol-P Glycerol
Glucose
Triacylglycerol

Fatty acyl CoA Fatty acid

gluconeogenesis
Malonyl CoA
Pyruvate

Acetyl CoA Ketone bodies

TCA cycle
Oxidation of fatty acids
• Central energy-yielding
pathway in animals.

=O • Generates acetyl-CoA
CH3-C-CoA

• Generates electrons –
which pass through the
respiratory chain driving
ATP synthesis.
 Sources of fatty acid fuels
1. de novo synthesis
• Fatty acids may be synthesized and are
converted to triacylglycerols

• Made in liver and exported to muscle or fat cells

• In muscle, used as fuel; In fat cells, stored as

droplets
 Sources of fatty acid fuels
2. diet
• Diet – on average 40% or more of the daily
energy requirement of humans is supplied in
the diet – in the form of triacylglycerols

• Stored in cells as lipid droplets

 Triacylglycerols

Extremely suitable as a energy storage

molecule:

1. Contain highly reduced hydrocarbon

chains with an energy more than twice
that of the same weight of carbohydrate or
protein
2. Extremely insoluble in water : less heavy
than hydrated molecules, such as
carbohydrates.
3. Chemically inert and so can be stored in
large quantity in cells.
Absorption of dietary fat
 Dietary fats are absorbed in the small
intestine

• Ingested triacylglycerols
are converted from
insoluble fat particles to
finely dispersed micelles

• Bile salts are amphipathic

cholesterol derivatives
made in the liver and
stored and released by the
gall bladder perform this
function
• Micelle formation allows lipid Ingested
triacylglycerols
molecules to be accessible to
water-soluble lipases (secreted by Lipases
pancreas)
Mono- and di-
acylglycerols,
free fatty acids,
glycerol
 The products of lipase action diffuse into the epithelial cells lining the intestinal surface
•

Then they are reconverted to triacylglycerols and packaged with dietary cholesterol and specific lipoprotein aggregates called chylomicrons.
•
 Molecular structure of a chylomicron
• The surface is a layer of
phospholipids, with
head groups facing the
aqueous phase.
• Triacylglycerols are in
the interior make up
more than 80% of the
mass.
• Apolipoproteins – lipid
binding proteins
• Chylomicrons move through the
lymphatic system, enter the blood
stream and are carried to muscle
and adipose tissue.

• In the capillaries of these tissues,

the extracellular enzyme
lipoprotein lipase hydrolyze
triacylglycerols to fatty acids and
glycerol, which are taken up by
cells in the target tissues.
• In muscle, the fatty
acids are oxidized
for energy.
• In adipose tissue,
they are esterified for
storage.
 Fate of dietary fat
• Dietary fat may be utilized immediately or
stored in adipocytes
• In fed state, fatty acid synthesis occurs and
the products are also stored in adipocytes
• In starved state, these stored forms of fat are
mobilized
 The Mobilization of
triacylglycerols

Fatty acids
Triacylglycerols +
Glycerol

• Triacylglycerols stored in the adipose tissues are

mobilized and transported to tissues where fatty
acids can be oxidized for energy production.
What signals the mobilization of
stored triacylglycerols?
• Hormones signal the need for metabolic
energy.

• Hormones epinephrine and glucagon are

secreted in response to low blood glucose
levels
 Glucagon and epinephrine act by a
signal transduction mechanism

Capillary

• These hormones activate the enzyme adenylyl

cyclase in the adipocyte plasma membrane.
• Results in increased amount of the secondary
messenger cAMP.
• cAMP activates
hormone-sensitive
triacylglycerol lipase
• The fatty acids released
passed into the blood
where they bind to blood
protein serum albumin.

• the insoluble fatty acids

are carried to tissues,
dissociated from albumin
and transported into cells
to serve as fuels.
 The products of fat mobilization
• Triacylglycerols are broken down to
glycerol and fatty acid

• 95% of the biologically available energy of

triacylglycerols resides in their long-chain
fatty acids
• 5% is contributed by the glycerol moiety
 Entry of glycerol into the glycolytic
pathway

• The glycerol released is

phosphorylated by glycerol
kinase to glycerol 3-
phosphate
• oxidized to dihydroxyacetone
phosphate
• then converted to
glyceraldehyde 3-phosphate,
which is oxidized via
glycolysis
 Fatty acids are activated and
transported into mitochondria
• FA oxidation enzymes
are located in
mitochondria
• Free FA cannot pass
directly through the
mitochondrial
membranes
 1. Activation of fatty acid by CoA

• Acyl CoA synthase

Fatty acid + CoA + ATP fatty acyl-CoA +AMP + PPi

 2. Esterification to carnitine

• Then transferred to carnitine

Fatty acyl-CoA + carnitine Fatty acyl- carnitine + CoASH

• The fatty acyl group is transferred to
carnitine by carnitine transferase I on the
outer face of the inner membrane
• The fatty acyl-carnitine ester then enters the
matrix through the acyl-carnitine/carnitine
transporter
 3. Esterification to CoA
• The fatty acyl group is enzymatically transferred from
carnitine to coenzyme A by carnitine acyltransferase II
• Regenerates fatty acyl-CoA and free carnitine
 Mitochondrial oxidation of fatty
acids takes place in three stages
• These stages result in massive ATP
production
 Stage 1.
β -oxidation:
 Oxidative removal
of successive two-
carbon units to form
acetyl-CoA starting
from carboxyl end of
the fatty acyl chain
 Also generates
NADH and FADH2
 Stage 2

• Acetyl groups of
acetyl-CoA are
oxidized to CO2 in
the TCA cycle
• NADH is also
generated from the
TCA cycle
 Stage 3
• The NADH and
FADH2 produced
mitochondrial
respiratory chain
 The β -oxidation of saturated
fatty acids has four basic steps

• The β -oxidation sequence is a

mechanism of breaking stable bond
between methylene (-CH2-) groups.
• The first three steps create a bond that is
more easily broken.
 Step 1
• Dehydrogenation of fatty acyl-CoA produces a double
bond between C-2 and C-3
• This double bond is in the trans configuration
• Yields FADH2
 Step 2
• Hydration - water is added to the double bond
to form β -hydroxyacyl-CoA.
 Step 3
 β -hydroxyacyl-CoA is dehydrogenated
(oxidized) to form β –ketoacyl-CoA.
(yields NADH + H+)
 Step 4
 β –ketoacyl-CoA reacts with free
coenzyme A to split off the carboxyl-
terminal two-carbon fragment of the
original FA as acetyl CoA.
The products for one pass through β -
oxidation

Palmitoyl-CoA + CoA + FAD + NAD+ + H20

myristoyl-CoA + acetyl-CoA + FADH2
+ NADH + H +

Myristate is a C14 fatty acid

 The four steps are repeated to
yield acetyl-CoA and ATP
• The myristoyl-CoA can go through another
set of four β -oxidation reactions to yield a
second molecule of acetyl-CoA and lauryl-
CoA (C-12).
 The complete β -oxidation of
palmitate

Palmitoyl-CoA + 7CoA + 7FAD + NAD+ + 7H20

8acetyl-CoA + 7FAD2 + 7NADH + 7H+
 The fate of FADH2 and NADH
• Each molecule of FADH2 ---2 molecules of
ATP .
• Each molecule of NADH delivers a pair of
electrons---3 molecules of ATP.
 The fate of acetyl-CoA
• Each molecule of acetyl-CoA can be
oxidized to CO2 and H2O by the TCA cycle
to yield 12 molecules of ATP.
 The overall ATP yield

Palmitoyl-CoA + 7CoA + 7FAD + NAD+ + 7H20

8acetyl-CoA + 7FAD2 + 7NADH + 7H+

• becomes
Palmitoyl-CoA + 23O2 + 108Pi + 131ADP
CoA + 131ATP + 16CO2 + 23H2O
 Oxidation of special fatty acids
• Mono and polyunsaturated fatty acids
• Odd chain fatty acids
Oxidation of unsaturated fatty acids
requires additional reactions

• Most fatty acids in triacylglycerols and phospholipids

are unsaturated.
• Being in the cis position these double bonds cannot
be acted upon by the β -oxidation enzymes
• By the action of two auxiliary enzymes such
substrates may be broken down
 Oxidation of a monounsaturated
fatty acid

• Example: oleic acid

• Requires enoyl-CoA
isomerase to reposition the
double bond.
• Converting the cis isomer
to a trans isomer, a normal
intermediate in β -
oxidation
 Oxidation of a polyunsaturated
fatty acid

• Example, linoleic
acid
• The first step is the
same as that
described above
• Complete oxidation requires
a second auxiliary enzyme, a
NADPH dependent reductase
that removes an unsaturated
bond

• The isomerase is required to

convert the double bonds
from a cis to a trans
configuration
 Complete oxidation of odd-
number fatty acids

• Odd numbered lipids are present in

plants and marine organisms
• Oxidized as even chain but end up
with proprionyl-CoA
 Proprionate metabolism

• Proprionyl-CoA is
carboxylated to
form D-methyl-
malonyl-CoA
• D-methyl-malonyl-
CoA is epimerized to
its L-stereoisomer

• L-methyl-malonyl-
CoA is converted to
succinyl-CoA, which
can enter TCA cycle.
 Fatty acid oxidation is tightly
regulated

• Fatty acid oxidation is regulated so it

occur only when the need for energy
requires it
 Two pathways fatty acyl-CoA in liver
Cytosol Mitochondria
Fatty acid Fatty
synthesis acid
oxidation

• The pathway taken Carnitine

depends on the rate Fatty acid
of transfer of long- transporter
chain fatty acyl-CoA
into the
mitochondria Triacylglycerols
and
phospholipids
Malonyl-CoA initiates fatty acid synthesis
Glycerol-P

Glucose
Triacylglycerol
• Malonyl-CoA is the first
intermediate in the cytosolic
biosynthesis of long-chain Fatty acyl CoA
fatty acids from acetyl-CoA
Malonyl CoA
Pyruvate
• Excess glucose that cannot
be oxidized or stored as
glycogen is converted in the Acetyl CoA
cytosol into FA for storage
as triacylglycerols TCA cycle
Malonyl-CoA inhibits carnitine transferase I
Cytosol Mitochondria
Fatty acid
synthesis Fatty
acid
Malonyl-CoA oxidation

Carnitine
Fatty acid
transporter

Triacylglycerols
and
phospholipids
 Fed state
Glycerol-P Glycerol
Glucose
Triacylglycerol

Fatty acyl CoA Fatty acid

Carnitine
transporter
Malonyl CoA
Pyruvate

Acetyl CoA

Insulin, citrate
TCA cycle
 Starved state
Glycerol-P Glycerol
Glucose
Triacylglycerol Glucagon/
epinephrine
gluconeogenesis

Fatty acyl CoA Fatty acid

Carnitine
transporter
Malonyl CoA
Pyruvate

Acetyl CoA Ketone bodies

TCA cycle
 Ketone bodies

• The acetyl-CoA formed in the

liver during β -oxidation can
have two fates:
1. Enter the TCA cycle
2. Converted to “ketone bodies”
– acetone, acetoacetone and
β -hydroxybutyrate – for
export to other tissues
 Ketone bodies formed in the liver
1. Condensation of two
molecules of acetyl-
CoA,
2. The resulting
acetoacetyl-CoA
condenses with acetyl-
CoA to form β -
hydroxy-β -
methylglutaryl-CoA
(HMG-CoA)
3. Cleavage of HMG-CoA yields acetyl-CoA
and acetoacetate.
4. Reduction of acetoacetate yields D-β -
hydroxybutyrate (do not confuse with L-
β -hydroxybutyrate of the b-oxidation
pathway).
5. Acetoacetate is easily decarboxylated (may
be spontaneously or enzymatically) to
acetone and CO2.
 Ketone bodies are exported to
other organs
• Acetone, produced in smaller quantities
than the other ketone bodies, is exhaled

• Acetoacetate and β -hydroxybutyrate are

transported in the blood to tissues other than
the liver
 Ketone bodies as fuels
 β -hydroxybutyrate may
be converted to acetyl-
CoA.

 The acetyl-CoA is
Oxidized in the TCA
cycle to provide much of
the energy required by
tissues
 Ketone bodies are used under
starvation conditions
• The brain, which preferentially uses glucose as fuel, can
adapt to the use of acetoacetate or β -hydroxybutyrate
under starvation conditions, when glucose is
unavailable
Intertissue relationships during starvation
 Summary of lipid metabolism
• Sources of triacylglycerols – diet and stored in
adipocytes
• Route taken by dietary triacylglycerols to muscle or
fat cells
• Mobilization of triacylglycerols is initiated by
hormones – epinephrine and glucagon
• The products of mobilization are free fatty acid and
glycerol, both are used for energy production
 Summary of lipid metabolism
• Carnitine transporter mediates entry of fatty acids
into mitochondria
• β -oxidation of fatty acids has four basic steps –
essentially fatty acid synthesis in reverse
• β -oxidation generates acetyl-CoA, NADH and
FADH2 ATP
• Ketone bodies serve as fuel molecules under
starvation conditions