Lipid Metabolism

in Lehninger Principles of Biochemistry, 5th Edition

Subject Chapter Reading

Lipid Structure & Chem (Review) 10 pp343-350,355,359
Lipid Degradation 17 pp 647-665
Ketogenesis 17 pp 666-668
Fatty acid Synthesis 21 pp 805-817
Integration 23 various
Triglyceride/Phospholipid Biosynthesis 21 pp 820-831
Cholesterol/Steroid metabolism/ 21 pp 831-845
Lipoproteins
 Lipid Classes (Review)
1. Simple fatty acids
- saturated
-unsaturated
2. Mono, di- and triglyceride
3. Glycerolphospholipids
(phosphoglycerides)
e.g. lecithin (phosphatidyl choline)
4. Sphingolipids
5. Steroids
6. Prostaglandins
7. Lipid Vitamins
 Lipid Catabolism
Triglyceride Glycerol + Fatty Acids
Glycolysis
Sat Unsat Odd Fatty Acids
TCA
Propionyl-CoA Succinyl-CoA

ACETYL-CoA
CO2

HMG CoA ACETYL-CoA

Ketone Bodies
 Lipid Anabolism(Biosynthesis)

ACETYL-CoA HMG-CoA Cholesterol

Steroids
Bile Acids
Fatty acids Vit D
Prostaglandins

Sphingolipids
Phospholipids Triglycerides
 General Triglyceride Catabolism
- Fatty acids in triglycerides (TG) are the major
source of energy in the body. TG are stored in
adipocytes or are found in dietary fat.
- 40% of daily energy requirement of people in
industrial countries is met with TG.
- Main tissues using fatty acids as energy source are
liver, heart and resting skeletal muscle
- A 70kg male has about 7 kg of lipid. A 65 kg
female has about 14 kg of lipid.
This is adequate energy needs for about a
month of starvation.
-There are carbohydrate stores for only
about a day.
-Carbohydrate and protein yield about 4 kcal
energy/g, whereas, triglycerides yield about 9
kcal of energy/g.
- Excess dietary carbohydrate is converted into
TG.

-Two reasons for using fat as an energy storage

form are its high energy yield/g and since fat is
very hydrophobic, fat can be stored in a compact
form.
Utilization of Dietary Fats
♦ Bile salts and pancreatic lipases in the small
intestines lead to the breakdown of dietary
fats to fatty acids, glycerol, mono- and
diglycerides for absorption into the intestine.

♦ These absorbed products are used to re-

synthesize triglycerides (TG) in the intestinal
cells and the TG are then distributed to
various tissues in the body by lipoproteins.
 Mobilization of Fats in Adipocytes (Fat Cells)
-Within the fat cells in peripheral tissues, lipids
are stored in oil drops.
-When needed, the triglycerides are broken down
by a different lipase called the hormone
sensitive lipase.
lipase
O lipase
O CH2O R CH2OH RCO2 _
R O H O
+ H2O HO H + RCO2 _

CH2O R CH2OH RCO2 _

triacylglycerides glycerol fatty acids

 Hormone Sensitive Lipase
1-Hydrolyzes
triglycerides to
glycerol and free
fatty acids
2-Hormonally
regulated
3-Provides mechanism
of liberating stored
fatty acids so they
can be distributed
to other tissues.

4-Albumin carries fatty

Lehn fig 17-3 acids in bloodstream
Regulation of Hormone Sensitive Lipase
Epinephrine or Glucagon

Insulin - stimulates adenylyl cyclase

cyclic AMP
stimulates protein kinase

inactive protein active protein kinase A

kinase A

ATP ADP

inactive lipase mobilized & active lipase- P

TG glycerol + fatty acids

 Glycerol Metabolism
CH2OH glycerol CH2OPO3 _
HO H + ATP kinase HO H + ADP
CH2OH CH2OH

glycerol–3-phosphate
glycerol NAD + NADH
glycerol phosphate
dehydrogenase
NADH NAD +

CH2OPO3 _

glycolysis O
CH2OH
dihydroxyacetone phosphate
Lehn fig 17-4 modified
 *Fatty Acid Catabolism*
Fatty Acid Activation
Fatty acids enter the cell as the free fatty
acid and must be activated into a fatty
acyl CoA to enable subsequent metabolism.

Fatty acyl CoA synthetase (thiokinase)

O O
RCO- + ATP + CoASH RC-SCoA + AMP
+ PPi
fatty acyl-CoA
 Two Steps in Overall Rxn
O O O
1. R-C-O- + ATP RCO-P-O- adenosine + PPi
O-
fatty acid fatty acyl adenylate 2Pi
(enzyme bound)

O O
2. RCO-P-O-adenosine + CoASH
O- O
AMP RC-SCoA +AMP
fatty acyl CoA
 Transport of Fatty Acids
into the Mitochondrion
♦ Fatty acids are stored in cytosol. Fatty acids
must be transported into the mitochondria,
where oxidative metabolism occurs.

♦Transport of fatty acids into the mitochondrion

requires carnitine and carnitine acyl
transferases.

carnitine
Carnitine Acyl Transferase I and II

Lehn fig 17-6

 β- Oxidation
1.oxidation to
double bond
between α and β
carbon
2. hydration across
double bond

3. oxidation to ketone
4. thiolysis releasing
acetyl-CoA and
shortened
(by two) acyl-CoA
 -Oxidation (cont’d) C16

• Two carbon groups are

oxidation
removed successively from the
carboxyl terminal
end of the fatty acid to give 7 rounds
acetyl-CoA.
hydration
• -Oxidation is a series of
oxidation, hydration and
cleavage reactions centered oxidation
around the  (3) carbon of the
fatty acid.

• Fatty acids are degraded by the thiolysis

release of C2 fragments as
acetyl-CoA by  -oxidation.
Lehn fig 17-8
Energy Yield from Oxidation of
Fatty Acids
Subsequent oxidation of the products results in the
yield of 17 ATPs per round of -oxidation.

FAD FADH2 2 ATPs

NAD+ NADH 3 ATPs

acetyl-CoA TCA cycle 12 ATPs

 Yield from Fatty Acid Oxidation
Fatty acid degradation occurs in the mitochondria and
yields acetyl-CoA and reduced cofactors via -oxidation.
Subsequent oxidation of these products by the TCA cycle
and the electron transport pathway yields much ATP.

Palmitoyl-CoA+7 CoASH+7 FAD + 7NAD+ + 7 H2O

8 acetyl-CoA + 7 FADH2 + 7 NADH + 7H+

8x12= 96 ATP 14 ATP 21 ATP

The 7 FADH2, 7 NADH and 8 acetyl-CoA,
therefore, yield 131 ATPs.

Since 2 ATP equivalents are necessary

to activate palmitate to palmitoyl CoA
(ATP AMP) the total energy yield from
palmitate oxidation is 129 ATPs = 8 ATPs/C.

Recall that 1 glucose 38 ATPs =

6.3 ATPs/C.

Clearly fatty acid oxidation yields

more storable energy.
 Odd Chain Fatty Acid Oxidation
Odd chain fatty oxidation yields the three
carbon propionyl-CoA. The metabolism of propionyl-
CoA involves the use of biotin and vitamin B12 and
results in the formation of succinyl-CoA.

propionyl-CoA carboxylase
CO2 + ATP [biotin]
O_
odd chain O O O _O O

fatty acids S COA H S COA O S COA

Beta- propionyl-CoA succinyl-CoA

oxidation n acetyl-CoA methylmalonyl-CoA
Lehn fig 17-11 modified
 Ketogenesis
♦Ketogenesis is a process by which acetyl CoA is
converted into ketone bodies (acetoacetic acid, β-
hydroxybutyrate and acetone).
acetone
♦Acetoacetic acid, and β-hydroxybutyrate are
distributed to peripheral tissues (muscle, brain)
to be used for ATP production.
♦HMG-CoA (hydroxymethylglutaryl CoA) is a
key intermediate in ketogenesis as well as sterol
synthesis.
 O
CH3 C CH3

acetone

Ketone O
O
Bodies CH3 C CH2
O

acetoacetate

OH
O
CH3 CH CH2
O
-hydroxybutyrate
Ketogenesis
1. condensation of two
acetyl-CoAs

2. add another acetyl

from acetyl-CoA

Lehn fig 17-19

3. split off
acetoacetate

4a. decarboxylate
to acetone
(normally
small amounts)
4b. reduce
 Fate of Ketone Bodies

LIVER fatty acid

oxidation
fatty acids acetyl-CoA acetoacetate -hydroxybutyrate

BLOOD

CO2 +ATP acetyl-CoA acetoacetate -hydroxybutyrate

citric acid
cycle succinyl-CoA
MUSCLE, BRAIN succinate Beta-ketoacyl-CoA
transferase
and KIDNEY acetoacetyl-CoA
 During untreated
diabetes, severe
dieting, and fasting,
ketogenesis and
gluconeogenesis
increase - there is
an over-production
of ketone bodies

Lehn fig 17-20

 Fatty Acid Biosynthesis
Overall reaction of fatty acid biosynthesis
14 NADPH + 8 acetyl CoA + 7 ATP
palmitate (C16) + 14 NADP+ + 8 CoA+ 7ADP +7 Pi +
H2O
This is a highly energy dependent process.
It occurs in the cytosol.
cytoso
The key enzyme which begins this process is acetyl CoA
carboxylase.
carboxylase It uses a biotin cofactor.
O O O
CH3C- SCoA + CO2 + ATP -
O-C-CH2-C- SCoA +
ADP + Pi
acetyl-CoA malonyl-CoA
♦ CO2 is required for fatty acid biosynthesis but is not found
in the final product. Malonyl-CoA is synthesized from
acetyl-CoA by acetyl-CoA carboxylase,
carboxylase which requires
ATP, CO2 and biotin.
♦Acetyl-CoA carboxylase, being the first enzyme in the
biosynthetic pathway is rate limiting.

♦ It is highly regulated
inactivated by phosphorylation (favored by glucagon)
activated by dephosphorylation (favored by insulin)
citrate also increases the activity allosterically
 Fatty Acid Synthase and Acyl Carrier
Protein
- Chain elongation occurs with fatty acid bound to a
carrier protein, acyl carrier protein, (ACP), which has
a prosthetic group related to CoASH.

- In eukaryotes ACP is actually part of a multifunctional

protein, fatty acid synthase that is responsible for fatty
acid chain synthesis.
 Lehn fig 21-5
modified

O O
|| ||
CH3C-SCoA + ACPSH CH3C-S-ACP +CoASH
malonyl-SCoA + ACPSH malonyl–S-ACP +
CoASH
 Fatty Acid Synthase
- Fatty acid synthase is a complex of enzymes that
adds C2 units to a growing fatty acid chain.
-It is a cyclic process that occurs with the fatty acid
covalently attached to ACP and requires reductive
reactions that use NADPH.

- The eukaryotic fatty acid

synthase is a homodimer of a
polyfunctional protein with
enzymes and carrier protein in
one polypeptide unit.
Fatty Acid Synthase
(cont’d)
malonyl attached to ACP,
acetyl to SH group.
acetyl displaces carboxylic
acid (as CO2),adding two
carbons.
ketone is reduced to hydroxide
using NADPH

Lehn fig 21-2 &21-3

Dehydration leaving a double bond

The double bond is hydrogenated

(reduced) using NADPH again

The acyl group is transferred

from the ACP to the SH
(not shown)

Lehn fig 21-2

First part is the set of
rxns just discussed Repeat: transferring butyryl to
Note: butyryl on SH acetyl on ACP and transferring
product back to SH
Process is
repeated
several times

After all necessary rxns,

product, palmitic acid,
is released
Lehn fig 21-4
 Rxn for palmitate synthesis is:
Acetyl-CoA + 7 malonyl-CoA + 14+ NADPH + 7 H+
palmitate + 7 CO2 + 14 NADP + 8 CoASH +
6H2O
But malonate is made from acetyl CoA

7 acetyl-CoA + 7 CO2 + 7 ATP 7 malonyl-CoA

+ 7 ADP + 7 Pi + 7 H+
So overall:
overall

8 acetyl-CoA + 7 ATP + 14 NADPH palmitate

+14 NADP+ + 8 CoASH + 6 H2O + 7 ADP + 7 Pi
The NADPH comes from the pentose phosphate
shunt. The source of acetyl-CoA is more complex.
Acetyl-CoA is required for fatty acid synthesis.

Acetyl-CoA is made in mitochondrial matrix.

Sources of acetyl-CoA are:

a) pyruvate dehydrogenase
b) breakdown of fatty acids
c) catabolism of ketogenic amino acids

-The mitochondrial membrane is impermeable to acetyl-

CoA
- So a shuttle system must be used for export these two
carbon fragments into cytosol.
 pyruvate
Citrate
synthase Citrate lyase
ATP, CO2
Acetyl CoA Acetyl CoA
ATP ADP

OAA citrate citrate OAA

NADH NADH

NAD+ NAD+
malate malate

mitochondrion membrane cytosol

Citrate-Malate Shuttle
 Citrate is the carrier of acetyl groups from the
mitochondrion to the cytosol. This is accomplished by
the action of citrate synthase and citrate lyase.

O O O
C O_ C O _ ATP ADP
C O_
Acetyl-CoA
O C O CH2 O C
C CH CH2
CH2 _O
CH2 Acetyl-CoA C
C
O O_ C O O_
oxaloacetate (OAA) citrate
O O _ oxaloacetate

citrate synthase citrate lyase

♦The oxaloacetate can be reduced to malate and
transported back into the mitochondrial matrix to be
oxidized to oxaloacetate.
♦ Instead cytosolic reducing equivalents for fatty acid
synthesis may be generated by the action of malic
enzyme.
O O
C O_
C O_
HO CH + NADP+ +NADPH + CO2+H+
O C
CH2
C CH3 pyruvate
O O_ malate
♦ Pyruvate is then transported into the mitochondrion
and carboxylated to oxaloacetate by pyruvate
carboxylase.
NADPH is also provided by the pentose phosphate shunt.
Differences Between Fatty Acid Synthesis and Oxidation

Parameter Oxidation Synthesis

Intracellular location Mitochondria Cytoplasm
Initial substrates Fatty acyl CoA Acetyl CoA,
malonyl CoA
Thioester linkage of CoASH Protein SH (ACP)
intermediates
Coenzymes FAD, NAD+ NADPH
Bicarbonate dependence NO YES
Energy state favoring processHigh ADP High ATP
Citrate activation NO YES
Acyl CoA inhibition NO YES
Highest activity Fasting and starvation Carbohydrate fed
Regulation of Lipid Metabolism

Metabolic regulation is achieved by

1. Hormonally stimulated phosphorylation-

dephosphorylation of regulated enzymes
2. Metabolic regulation by allosteric control
3. Control of Expression
Discuss
Integrative aspects of regulation
Key Regulatory Enzyme is
Acetyl CoA Carboxylase
Biotin
acetyl-CoA + ATP + HCO3- malonyl-CoA +
ADP + Pi + H+

♦ inactivated by phosphorylation (glucagon)

♦ activated by dephosphorylation (insulin)
♦ allosterically activated by citrate
 1a. Hormonal Control- Glucagon
Binding of glucagon to its plasma membrane
receptor results in the subsequent
phosphorylation of enzymes by protein kinases.
a. Acetyl CoA carboxylase ( inhibited)
b. Hormone sensitive lipase ( activated)
c. Fructose bisphosphatase-2 (active)
With the following results

a. Decreased fatty acid synthesis

b. Increased mobilization of stored fats
c. Increased gluconeogenesis
 1b. Hormonal Control- Insulin
Binding of the insulin to its plasma membrane
receptor results in the subsequent
dephosphorylation of enzymes by phosphoprotein
phosphatases.
a. Acetyl CoA carboxylase ( activated)
b. Hormone sensitive lipase ( inactivated)
c. Phosphofructokinase-2 ( activated)
With the following results

a. Increased fatty acid synthesis

b. Decreased mobilization of stored fats
c. Decreased gluconeogenesis
Regulation of Acetyl CoA Carboxylase
 Compare
low blood sugar, ↑gluconeogenesis increased fat
glucagon oxidation/low fatty
acid biosynthesis
high blood sugar, ↓gluconeogenesis increased fatty acid
insulin biosynthesis/low fat
oxidation

2. Regulation by Allosteric Control

♦ Palmitoyl-CoA inhibits fatty acid synthesis by

allosteric inhibition of acetyl CoA carboxylase

♦ Acetyl-CoA carboxylase is activated by citrate

♦ Carnitine palmitoyl transferase is inhibited by

malonyl CoA
 acetyl CoA
carboxylase
filaments
Lehn fig 21-11
Inhibition of Carnitine Acyl Transferase-I

Conditions favor synthesis- malonyl-CoA helps

retain fatty acids in cytosol
3. Changes of enzyme synthesis

a. fatty acid synthesis is induced

b. acetyl Co A carboxylase is induced

c. adipose lipoprotein lipase is induced (more

on this later)
 Enzymes, Proteins & Other Players in
Triacylglyceride & Fatty Acid Degradation
Hormone sensitive lipase
Albumin
Glycerol kinase
Glycerol-3-phosphate dehydrogenase
Fatty acyl-CoA synthetase (thiokinase)
carnitine
Carnitine acyl transferase I &II
Acyl-CoA dehydrogenase, hydratase,
OH acyl-CoA dehydrogenase
Acyl-CoA acetyltransferase
Propionyl-CoA carboxylase
in fat cells, hydrolyzes TG to release fatty
acids, controlled by glucagon, epinephrine
& insulin
carries fatty acids in blood stream
produces glycerol-3-phosphate
oxidizes G3PD to DHAP for glycolysis
uses ATP & CoA to activate fatty acids
small molecule used to transport fatty
acids into mitochondrion
adds fatty acyl to carnitine from fatty acyl
CoA & removes
dehydrogenates (oxidizes) fatty acyl CoA,
hydrates and oxidizes to ketone at beta
position
removes acetyl-CoA by attack of CoA
leaving shortened acyl CoA
adds CO2 to propionyl-CoA to form
methylmalonyl –CoA, uses ATP, biotin &
B12
Enzymes, Proteins & Other Players in Ketogenesis
& Utilization of Ketone Bodies
Ketone bodies acetoacetone, D-beta hydroxybutyrate and
acetone
Thiolase puts 2 acetyl CoAs together to form
acetoacetyl-CoA
HMG-CoA synthase adds a 3rd acetyl-CoA to produce HMG-
CoA; also in chol synthesis
D-Beta-hydroxybutyrate dehydrogenase oxidizes acetoacetate to D-beta-
hydroxybutyrate
Beta-ketoacyl CoA transferase exchanges acetoacetyl for succinyl in
succinyl-CoA
Diabetic ketosis large increase in levels of circulating ketone
bodies in untreated diabetes
Enzymes, Proteins & Other Players in Fatty Acid
Biosynthesis
Acetyl CoA carboxylase
Acyl carrier protein, ACP
Fatty acid synthase
Citrate-malate shuttle
Citrate synthase & citrate lyase
carboxylates acetyl-CoA to make
malonyl-CoA, uses ATP & biotin,
regulated & rate limiting
has prosthetic group, part of fatty acid
synthase, carries malonyl-CoA & acyl-
CoA
homodimer of large polypeptide,
synthesizes fatty acids
transports acetyl-CoA from mit to cytosol
produces acetyl-CoA carrier in mit &
releases acetyl CoA in cytosol