OXIDATION OF FATTY ACIDS:

KETOGENESIS
 LEARNING OBJECTIVES FOR TODAY:

1. To describe the processes by which fatty acids are transported in

blood and activated in the cytosol

2. To outline the beta-oxidation pathway by which fatty acids are

metabolized to acetyl CoA

3. To compute for total bioenergetics produced and consumed by

fatty acid beta-oxidation
LIPIDS
FATTY ACID BETA OXIDATION
 BETA OXIDATION OF FATTY ACIDS
Catabolic pathway of fats

Free fatty acids are converted to Acetyl CoA

Free/Nonesterified Citric Acid Cycle

or Unesterified Acetyl CoA and ETC
fatty acid
O2
 WHY DO WE NEED TO BREAKDOWN FATTY ACIDS
FURTHER?
Major energy reserve

Oxidation: 9 kcal/gm of fat > 4 kcal/gm of carbohydrates

Constitutes 11kg of 70kg of total body weight (84% of

stored energy)
 Carboxyl group
FATTY ACIDS
α-carbon
Basic structural unit of lipids
β-carbon

Relatively insoluble in water (the

more hydrocarbons are present,
the more hydrophobic)

Soluble in nonpolar solvents (ether

and chloroform) R-COOH
WHAT IS A GOOD SOURCE OF FATTY ACIDS IN THE BODY?
TRIACYLGLYCERIDES (TAG) IN STORES
Major energy reserve: 9 kcal/gm
of fat
Site of accumulation: cytoplasm of
adipose cells

Adipose tissue: specialized for the

synthesis, storage and mobilization
of lipids
 PROCESS OF BETA-OXIDATION
Can be divided into three (3) phases:

1. Breakdown of stored TAG and transport fatty acid from adipose

tissue to target cells (Lipolysis + fatty acid transport)

2. Fatty acid activation in Cytosol

3. Transport of activated fatty acids into mitochondria and

oxidative catabolism inside mitochondrial matrix
FIRST PHASE: LIPOLYSIS AND FATTY ACID TRANSPORT
 LIPOLYSIS
Glucagon, adrenalin, ACTH
↓
receptor activation
↓
protein kinase A
↓
phosphorylation of Perilipin
(when dephosphorylated it inhibits the
access of lipases to TG and DG
+
phosphorylation of HSL
(hormon-sensitive lipase)
activation
↓
CGI dissociates from Perilipin
then associates with ATGL (adipocyte
trigliceride lipase)
↓
TG→DG + fatty acid
↓
HSL: DG→MG + fatty acid
↓
MGL (monoacylglycerol lipase):
MG→G + fatty acid
FATTY ACID TRANSPORT Fatty acids are highly water-
insoluble
Fatty acid must be
transported in blood together
with a protein

1. Fatty acid esters

(Esterified)
2. Free fatty acids (Unesterified)
1. Fatty acid esters
- more than 90% of blood fatty acids are
esters
- e.g. TAG in chylomicrons, cholesterol
esters, phospholipids; they are transported in
blood with lipoprotein particles

2. Free fatty acids (Unesterified)

- less than 10%
- e.g. not bound to any other molecule
-must be transported together with albumin
 Fatty acids + Albumin

10 Fatty acids/albumin monomer

Blood Circulation

Skeletal muscle, heart muscle, renal cortex

Fatty acid activation, transport into the mitochondria, beta-oxidation

SECOND PHASE: ENTRY INTO CYTOPLASM
ENTRY INTO THE CYTOPLASM
FATTY ACID ACTIVATION
FA are converted to CoA thioesters
by FA acyl-CoA synthetase (FA
thiokinase)

FA thiokinase: found in outer

membrane of mitochondria BUT FA
activation occurs in cytosol

ATP dependent
ACTIVATION: FATTY ACID TO ACYL COA
THIRD PHASE: ENTRY INTO MITOCHONDRIA
 TRANSLOCATION OF ACYL COA
Acyl CoA from SCFA/MCFA
can traverse the outer
membrane and inner
mitochondrial membrane
directly

Long chain fatty acids are

converted to Carnitine for
the purpose of transport into
intermembrane space
INTERMEMBRANE SPACE: Carnitine Acyl Transferase I: Acyl
CoA  Acyl-carnitine

Translocase: Only recognizes acylcarnitine; it translocated

the carnitine structure through the inner membrane to the
matrix

MITOCHONDRIAL MATRIX: Carnitine Acyl Transferase II:

Acyl-carnitine  Acyl CoA
-in the matrix, fat is esterified back into CoA
SUMMARY
BETA-OXIDATION IN MITOCHONDRIAL MATRIX
 BETA-OXIDATION PROPER
A four-step process:
1) Oxidation by FAD-linked Acyl CoA dehydrogenase
2) Hydration by hydratase
3) Oxidation by NAD-linked dehydrogenase
4) Thiolysis by thiolase
 Three isoenzymes:

STEP I: OXIDATION -long-chain fatty acids (C 12-18)

-medium chain fatty acids (MCAD, 4-14)
-short-chain fatty acids (4-8)
MCAD deficiency is relatively frequent

The first reaction is oxidation

of acyl CoA by an acyl CoA
dehydrogenase to give α-β
unsaturated acyl CoA (enoyl
CoA)

FAD is used as the hydrogen

acceptor
 STEP II: HYDRATION

The second reaction is the

hydration of the double bond
to β-hydroxyacyl CoA
 STEP III: OXIDATION

The third reaction is the oxidation

of β-hydroxyacyl CoA to produce
β-ketoacyl CoA with a NAD-
dependent reaction
 STEP IV: THIOLYSIS
The fourth reaction is
cleavage of the two carbon
fragment by splitting the bond
between α and β carbons
using a thiolase enzyme

Reaction will repeat up until

all carbons of the FA is
removed
HOW MANY ACETYL COA
ARE YOU GOING TO
PRODUCE FROM PALMITIC
FATTY ACID (C16)?
The release of acetyl CoA leaves an acyl CoA molecule
shortened by 2 carbons

This acyl CoA molecule is the substrate for the next round of
oxidation starting with acyl CoA dehydrogenase

Repetition occurs until all the carbons of the original fatty acyl
CoA are converted to acetyl CoA

In the last round, a four carbon acyl CoA (butyryl CoA) is
cleaved into 2 acetyl CoA
BIOENERGETICS OF FA OXIDATION
Example: Palmitic Acid (16 Carbon fatty
acid)

1. Beta-oxidation of palmitic acid will be

repeated 7 cycles producing 8
molecules of acetyl CoA
2. In each cycle, FADH2 and NADH is
produced and will be transported to
the ETC
FADH2 = 1.5 ATP NADH = 2.5 ATP
NOTE: Some books will round off the mol
ATP generated (FADH2 = 2 ATP; NADH =
3 ATP)
So 7 cycles:
(FADH2 + NADH) x 7 cycles = (1.5 ATP + 2.5 ATP) x 7 = 28 ATP
(FADH2 + NADH) x 7 cycles = (2 ATP + 3 ATP) x 7 = 35 ATP
3. Each of acetyl CoA when oxidized through the citric acid cycle gives
10 or 12 ATPs

Recall: 1 acetyl CoA  Citric Acid Cycle will yield 3 molecules of

NADH, 1 molecule of FADH2 and 1 ATP/GTP
3 NADH molecules = 2.5 ATP x 3 = 7.5 ATP 3 ATP = 9 ATP
1 FADH2 molecules = 1.5 ATP x1 = 1.5 ATP 2 ATP = 2 ATP
1 ATP or GTP = 1ATP__ 1 ATP = 1 ATP
10 ATPs 12 ATPs
3 NADH molecules = 7.5 ATP x 8 = 60 ATP 9 ATP x 8 = 72 ATP
1 FADH2 molecule = 1.5 ATP x 8 = 12 ATP 2 ATP x 8 = 16 ATP
1 ATP/GTP molecule = 1 ATP x 8 = 8 ATP 1ATP x 8 = 8 ATP
80 ATP 96 ATP
10 ATPs x 8 = 80 ATPs
12 ATPs x 8 = 96 ATPs
4. 2 ATPs are utilized in the activation of fatty acid (It
occurs once)

Energy Gain = Energy produced – Energy Utilized

= 28 + 80 ATP – 2 ATP = 106 ATP
= 35 + 96 ATP – 2 ATP = 129 ATP
WHAT HAPPENS TO GLYCEROL?

Glycerol will be converted to Glycerol-3-Phosphate intermediate in the liver

REGULATION OF FATTY ACID BETA-OXIDATION
REGULATION
 REGULATION OF BETA-OXIDATION
Positive Effectors: Starvation and a general low-energy level
a) Low insulin and high glucagon (Low insulin:glucagon ratio)

Malonyl-CoA inhibits it because it is a reactant of fat synthesis

a) Inhibits Carnitine acetyltransferase, preventing transport of fats
into mitochondria
b) In conjunction with fat synthesis, so that newly made fats are
not broken down again
Negative effectors: General indicators of sufficient energy in
the cell
a) High insulin and low glucagon
b) High ATP
 LONG TERM REGULATION
PPAR (Peroxisome proliferator/activated receptors)
Nuclear receptors: transcription factors

PPARα: Muscle, adipose tissue and liver

-regulate transcription of fatty acid transporters, CPT I
and CPT II, and acyl CoA dehydrogenase
 CLINICAL CORRELATE
The most common genetic defect in fatty acid oxidation is Acyl-CoA
dehydrogenase deficiency

For the medium chain acyl CoA dehydrogenase

-1:40 mutation in one of the chromosomes
-1:10000 two mutant copies (disease manifestation in first years)

Symptoms: Hypoglycemia with decreased ketone body formation

(decreased fatty acid oxidation and gluconeogenesis in the liver)
Accumulation of lipids in the liver (hepatic ____)

Therapy: Frequent carbohydrate-rich meals

Carnitine Acyltransferase Deficiency: most common: CPT
II gene mutation (partial loss)
a) muscle weakness
b) serious: hypoglycemia with decreased ketone body
formation
KETONE BODY FORMATION
 FATTY ACID
GLUCOSE

GLUCONEOGENESIS
ß-oxidation

PYRUVATE ACETYL-CoA

OXALOACETATE
Ketone bodies

CITRATE
Fatty acid oxidation + lack of oxaloacetate

fasting
untreated diabetes
THREE KETONE BODIES
SYNTHESIS OF KETONE BODIES IN THE LIVER
KETONE BODIES AS FUELS
 OXIDATION OF KETONE
BODIES IN EXTRAHEPATIC
TISSUES

heart muscle
striatal muscle
kidney
brain
 SUMMARY ON KETONE BODIES

Ketone bodies are

important sources of
energy in heart muscle,
renal cortex
(preference to glucose,
1/3 of energy) and
brain (Glucose is main
fuel but in starvation
and DM, brain uses
acetoacetate
 KETONE BODIES
Fasting
Diabetes
High levels of ketone bodies in the blood
(KETOSIS)

Level of ketone bodies after an overnight fast: ~0.05 mM

2 days starvation: 2 mM (40-fold increase)
40 days: 7 mM