(Ch 17) Fatty Acid Catabolism

(b-oxidation)

• Digestion, Mobilization, and Transport of Fats

• Oxidation of Fatty Acids

• Ketone Bodies
 Fatty Acid Catabolism (b-oxidation)

• Greatest fraction of fuel for most organisms

and organs
– Vertebrates
– Muscle (including heart), liver
• Fat sources for energy
– Ingested
– Taken from stores (Adipocytes)
– Synthesized in liver from carbohydrates

under all physiological circumstance, b-oxidation in mammalian heart and liver,

provides as much as 80% of the energetic needs.
 Dietary Fats Are Absorbed in the Small Intestine

Because chylomicron is too big to move through capillaries as protein or

glucose, so it move through lymphatic system
 Molecular structure of a chylomicron

• The surface is a layer of phospholipids,

with head groups facing the aqueous
phase.

• Triacylglycerols sequestered in the

interior (yellow) make up more than
80% of the mass. 100 to 500 nm.

• Several apolipoproteins that protrude 80%

from the surface (B-48, C-III, C-II) act as
signals in the uptake and metabolism of
chylomicron contents.

• The diameter from 100 to 500 nm

Hormones Trigger Mobilization of Stored Triacylglycerols

Liver

Muscle
Tissue
Adipose Tissue

Neutral
lipid
 Hormones Trigger Mobilization of Stored Triacylglycerols

• The surface of chylomicron droplets is coated with perilipins, a family of proteins that
restrict access to lipid droplets, preventing untimely lipid mobilization

• When hormones signal the need for metabolic energy, triacylglycerols stored in adipose
tissue are mobilized (brought out of storage) and transported to tissues (skeletal muscle,
heart, and renal cortex) in which fatty acids can be oxidized for energy production

• Cyclic AMP–dependent protein kinase (PKA) phosphorylates perilipin A, and causes

hormone-sensitive lipase (HSL) in the cytosol to move to the lipid droplet surface, where it
can begin hydrolyzing triacylglycerols to free fatty acids and glycerol.

• PKA also phosphorylates hormone-sensitive lipase, doubling or tripling its activity.

• About 95% of the biologically available energy of triacylglycerols resides in their three long-
chain fatty acids; only 5% is contributed by the glycerol moiety
Entry of glycerol into the glycolytic pathway

aldolase
F 1,6 bis-p

gluconeogensis
 β-oxidation of fatty acids

• substrate: Fatty acyl-CoA

• product: n acetyl-CoA, n NADH + H+, n FADH2
• function: gain of energy from fatty acids
• Organ location: liver, skeletal muscles and other
tissues with expection to CNS
• regulatory enzyme: carnitine acyltransferase I
 Beta Oxidation of Fatty Acids

• Process by which fatty acids are degraded by

removal of 2-C units
• b-oxidation occurs in the mitochondria matrix
• The 2-C units are released as acetyl-CoA, not free
acetate
• The process begins with oxidation of the carbon
that is "beta" to the carboxyl carbon, so the
process is called "beta-oxidation"
 Fatty acids must first be activated by formation
of acyl-CoA
• Acyl-CoA synthetase condenses fatty acids with CoA, with
simultaneous hydrolysis of ATP to AMP and PPi
• Formation of a CoA ester is expensive energetically
• Reaction just barely breaks even with ATP hydrolysis DGo’ATP
hydroysis = -32.3 kJ/mol, DGo’ Acyl-CoA synthesis +31.5
kJ/mol.
• But subsequent hydrolysis of PPi drives the reaction strongly
forward (DGo’ –33.6 kJ/mol)
Activation
 Import of acyl-CoA into mitochondria
• B-oxidation occurs in the mitochondria,
requires import of long chain acyl-CoAs
• 1. Acyl-CoAs are converted to acyl-
carnitines by carnitine acyltransferase.
1 2
• 2. A translocator then imports Acyl
carnitine into the matrix while
simultaneously exporting free carnitine
to the cytosol
• 3. Acyl-carnitine is then converted back
to acyl-CoA in the matrix.
• 4. The carnitine-mediated entry process
is the rate limiting step for oxidation of
fatty acids in mitochondria.
4 3
 Carnitine Shuttles
Transport

The fatty acids with 12 or fewer carbons enter mitochondria without the help of membrane
transporters.
But those with 14 or more carbons, which constitute the majority of the FFA obtained in the
diet or released from adipose tissue, cannot pass directly through the mitochondrial
membranes—they must first undergo the three(acyl CoA synthetase one of them) enzymatic
reactions of the carnitine shuttle.
FA with14 or more carbons
 Deficiencies of carnitine or carnitine transferase or
translocator activity are related to disease state

• Symptoms include muscle cramping during exercise,

severe weakness and death.
• Affects muscles, kidney, and heart tissues.
• Muscle weakness related to importance of fatty acids
as long term energy source
• People with this disease supplement diet with medium
chain fatty acids that do not require carnitine shuttle
to enter mitochondria.
 b-oxidation
• Strategy: create a carbonyl group on
the β-C
• First 3 reactions do that; fourth
cleaves the "β-keto ester"
• Products: an acetyl-CoA and a fatty
acid two carbons shorter
B-Oxidation
 Acyl-CoA Dehydrogenase

• Oxidation of the C-C bond

• Mechanism involves
proton abstraction
(removal), followed by
double bond formation (in
trans configuration) and
hydride removal by FAD
• Electrons are passed to an electron transfer flavoprotein, and then
to the electron transport chain.
 Enoyl-CoA Hydratase
• Adds water across the double bond.
• Uses substrates with trans-D2-and cis D2
double bonds (important in B-oxidation of
unsaturated FAs).

• With trans-D2 substrate forms L-isomer,

with cis D2 substrate forms D-isomer.

• Normal reaction converts trans-enoyl-CoA

to L-β-hydroxyacyl-CoA .

• note that naturally occurring unsaturated

fatty acids are in cis configuration
 Hydroxyacyl-CoA Dehydrogenase
• Oxidizes the 3-Hydroxyl
Group to keto group

• This enzyme is completely

specific for L-hydroxyacyl-
CoA

• D-hydroxylacyl-isomers are
handled differently
• Produces one NADH
 Thiolase
• Nucleophillic sulfhydryl group of
CoA-SH attacks the b-carbonyl
carbon of the 3-keto-acyl-CoA.

• Results in the cleavage of the Ca-

Cb bond.
• Acetyl-CoA and an acyl-CoA (-) 2
carbons are formed.

• Note:4 hydrogen atoms were

removed by dehydrogenases
Energy Yield
 b-oxidation
• B-oxidation of palmitate (C16:0) yields 106 molecules of ATP
• C 16:0-CoA + 7 FAD + 7 NAD+ + 7 H20 + 7 CoA à 8 acetyl-CoA + 7
FADH2 + 7 NADH + 7 H+

2.5 ATPs per NADH = 17.5

1.5 ATPs per FADH2 = 10.5
10 ATPs per acetyl-CoA = 80
Total = 108 ATPs

• 2 ATP equivalents (ATPà AMP + PPi, PPi à 2 Pi) consumed

during activation of palmitate to acyl-CoA
• Net yield = 106 ATPs
Oxidation of unsaturated fatty acid

• Mitochondria

• Isomerase: cis → trans

• Epimerase: D (-) → L (+)

 b-oxidation of odd
chain fatty acids
• Odd chain fatty acids are less common
(naturally lipids are even number)
• Formed by some bacteria in the
stomachs of ruminants and the human
colon.
• B-oxidation occurs pretty much as
even chain fatty acids until the final
thiolase cleavage a five-carbon fatty
acid which results in acetyl-CoA and a
3 carbon acyl-CoA (propionyl-CoA).
Special set of 3 enzymes are required
to further oxidize propionyl-CoA
• Final Product succinyl-CoA enters TCA
cycle
Take care of the enzymes and its coenzymes and cofactors
Oxidation of propionyl-CoA

propionyl-CoA
Carboxylase (biotin)
Epimerase
Mutase (VB12)

succinyl-CoA
enters TCA cycle
Oxidation of a monounsaturated fatty acid.
Oleic acid This product cannot serve as a substrate for enoyl-CoA
hydratase, which acts only on trans double bonds.

The auxiliary enzyme are needed

for _ oxidation of the common
unsaturated fatty acids:
an isomerase and a reductase
enoyl-CoA
Isomerase

isomerizes the cis--enoyl-CoA to

the trans-enoyl-CoA, which is
converted by enoyl-CoA
hydratase into the corresponding
L-hydroxyacyl-CoA
(trans-2-dodecenoyl-CoA
 b-oxidation of unsaturated fatty acids
• b-oxidation occurs normally for 3 rounds
until a cis- Δ3-enoyl-CoA is formed.
• Acyl-CoA dehydrogenase can not add
double bond between the a and b
carbons.
• Enoyl-CoA isomerase converts this to
trans- Δ2 enoly-CoA
• Now the b-oxidation can continue on w/
the hydration of the trans-D2-enoyl-CoA
• One cis double bond, Odd numbered
double bonds handled by isomerase
 Two cis d.b.’s
—Takes a new reductase, too…
—And then another isomerase…
Oxidation of a polyunsaturated linoleic acid fatty acid.
 Synthesis of ketone bodies (ketogenesis)

• substrate: fatty acetyl-CoA

• product: acetoacetate, 3-hydroxybutyrate, acetone
• function: energy substrate for extrahepatal tissues
• subcelullar location: matrix of mitochondria
• organ location: liver

Excessive production of ketone bodies is typical during starvation

or diabetes mellitus:
↑ lipolysis → ↑ FA → β-oxidation of FA → excess of acetyl-CoA
→ ↑ ketogenesis
 Ketone Bodies

are water-soluble fuels normally exported by the liver

but overproduced during fasting or in untreated
diabetes mellitus
 Ketone Bodies
• A special source of fuel and energy for certain tissues
• Produced when acetyl-CoA levels exceed the capacity of the TCA cycle
(depends on OAA levels)
• Under starvation conditions no carbohydrates to produce anpleorotic
intermediates
• Some of the acetyl-CoA produced by fatty acid oxidation in liver
mitochondria is converted to acetone (small amount), acetoacetate
and b-hydroxybutyrate
• These are called "ketone bodies“ soluble in blood and urine.
• Source of fuel for brain (adaptation under starvation), heart and
muscle
• Major energy source for brain during starvation
• They are transportable forms of fatty acids!
In liver Ketone Bodies
In periphery

Liver lacks this enzyme

therefore is unable
to use ketone bodies as fuel

TCA cycle
 Use of ketone bodies by the extrahepatal
tissues
• acetoacetate and 3-hydroxybutyrate are reconverted
to acetyl-CoA (→ citric acid cycle)

• is located in matrix of mitochondria of the peripheral

tissues

• is significant in skeletal muscles, heart and also in the

brain if lack of Glc occurs
 Ketone Bodies and Diabetes
• Lack of insulin related to uncontrolled fat
breakdown in adipose tissues
• Excess beta-oxidation of fatty acids results in
ketone body formation.
• Can often smell acetone on the breath of
diabetics.
• High levels of ketone bodies leads to condition
known as diabetic ketoacidosis.
• Because ketone bodies are acids, accumulation
can lower blood pH.
 Causes for ketosis

• Severe diabetes mellitus

• Starvation
• Hyperemesis (vomiting) in early
pregnancy
Hepatocyte

Acetoacetate,
β-hydroxybutyrate,
acetone Ketone bodies
exported as
Ketone energy source
CoA for heart,
body
formation skeletal muscle,
Fatty Citric kidney, and
acids Acetyl-CoA brain
Acid cycle
β-oxidation

oxaloacetate
gluconeogenesis

Glucose Glucose exported

as fuel for tissues
such as brain
Plasma concentrations of metabolic fuels
(mmol/L) in the fed and starving states
Ketosis consists of ketonemia, ketonuria and
smell of acetone in breath
 w-oxidation
• Minor pathway, but when oxidation is defective (because of
mutation or a carnitine deficiency, for example) it becomes more
important
• The enzymes located In the endoplasmic reticulum of liver and
kidney.
• 3 additional enzymes.
• substrates are fatty acids of 10 or 12 carbon atoms.
The first step introduces a hydroxyl group onto the
carbon . The oxygen for this group
comes from molecular oxygen (O2) in a complex reaction
that involves cytochrome P450 and the electron
donor NADPH catalyzed by mixed-function oxidases.

Two more enzymes now act on the carbon: alcohol

dehydrogenaseoxidizes the hydroxyl group to an
aldehyde, and aldehyde dehydrogenase oxidizes
the aldehyde group to a carboxylic acid, producing a
fatty acid with a carboxyl group at each end.

At this point, either end can be attached to

coenzyme A, and the molecule can enter the
mitochondrion and undergo oxidation by the normal
route. In each pass through the -oxidation pathway,
the “double-ended” fatty acid yields dicarboxylic
acids such as succinic acid, which can enter the
citric acid cycle, and adipic acid