Topic 8 : Metabolism in Liver 2

• Objectives
 Synthesis and Storage of fatty acids
 Comparison of synthesis and breakdown
of fatty acids
 Synthesis of triacylglycerol
 Ketone body formation
 Utilisation of ketone bodies

Topic 8 : Metab. Liver 2 1

 Fatty Acid Synthesis
 Early studies showed that synthesis of fatty acids not a
reversal of degradation, although pathways very similar.
 The two processes occur in different
compartments of cells.
 Requirement for bicarbonate in synthesis but not
incorporated into product.
• Fatty acid synthesis from acetyl-CoA and malonyl-
CoA occurs by a series of reactions, catalyzed in bacteria
by seven different enzymes.
• In eukaryotes, synthesis catalysed by Fatty Acid
Synthase System (FAS).
• NADPH serves as electron donor in two reactions
involving substrate reduction. The NADPH is produced
mainly by the Pentose Phosphate Pathway.
Topic 8 : Metab. Liver 2 2
 Conversion of Carbohydrate to Fat
 Diet of humans mainly carbohydrate.
 Storage of carbohydrate limited because
more “bulky”.
 Instead stored as fat; weight for weight fats
yield more energy than carbohydrates.
 Excess carbohydrate converted to fats;
hence requirement for conversion of
carbohydrate to fats.
 Central metabolite is acetyl CoA, formed
from oxidative decarboxylation of
pyruvate and β-oxidation of fatty acids.
Topic 8 : Metab. Liver 2 3
Acetyl Co A as key Intermediate between fat
and carbohydrate metabolism

 Carbohydrates can be
converted to fats but
not vice-versa except
propionyl CoA and
glycerol.
 PDH virtually
irreversible →
pyruvate cannot be
formed from acetyl
Topic 8 : Metab. Liver 2
CoA 4
 Transfer of Acetyl units and Reducing
Equivalents for Fatty Acid Synthesis
 Acetyl CoA is produced in the mitochondria whereas fatty acid
synthesis occurs in the cytosol
 Acetyl CoA like all CoASH derivatives cannot penetrate the
mitochondrial membrane.
 Shuttle mechanism exists which not only transports the acetyl units
but also provides mechanism to supply some of the NADPH

1 = Citrate synthase
2 = Citrate lyase 2
1
3 = Malate dehydrogenase
4 = Malic enzyme
5 = Pyruvate carboxylase 3 3

5 4
Topic 8 : Metab. Liver 2 6
 Acyl Carrier Protein
 In fatty acid oxidation, all the intermediates are linked to a carrier
molecule, coenzyme A (CoASH)
 Fatty acid synthesis involves a carrier molecule known as acyl
carrier protein (ACP)

7
 Formation of Acetyl- and Malonyl–acyl
carrier protein (ACP)
• First committed step
in fatty acid synthesis
is carboxylation of
acetyl CoA to form
malonyl CoA
• Catalysed by acetyl
CoA carboxylase, an
enzyme which is
allosterically
stimulated by citrate,
contains biotin as
prosthetic group.
• Elongation steps in FA synthesis involve all intermediates linked
to terminal sulphydryl group of phosphopantetheine of ACP.8
 Reactions of FA Synthesis
• Cycle of reactions involve
 Condensation of acetyl–ACP
and malonyl–ACP with release
of CO2.
 Reduction with NADPH.
 Dehydration to yield a trans
double bond.
 Reduction with NADPH.
• First cycle yields butyryl–ACP
(C4).
• Cycle repeats with malonyl–ACP
adding two-carbon units.
• Chain elongation until palmitoyl-
ACP (C16).
• Molecule not accepted by acyl –
malonyl–ACP condensing enzyme.
• Hydrolysed by thioesterase to give
palmitate and ACP.
9
Topic 8 : Metab. Liver 2 11
 Enzymes in Fatty Acid Synthesis
• Overall reaction
8Acetyl CoA + 7ATP + 14NADPH + 6H+ → Palmitate
+ 14 NADP+ + 8CoASH + 6H20 + 7ADP + 7 Pi
• In prokaryotes, reactions of FA synthesis carried out
by separate enzymes.
• In eukaryotes, enzymes present in a single
polypeptide chain, multifunctional enzyme complex
called fatty acid synthase (FAS).
• Swinging arm of phosphopantetheine (PPT) brings
acyl groups into contact with active sites of FAS
• Fatty acid synthase complex exits as a dimer and
arranged in such a way as to increase the efficiency of
FA synthesis. Topic 8 : Metab. Liver 2 12
 Fatty Acid Modification
Synthesis of odd-chain fatty acids
• Propionyl –ACP (3-C) used as primer – occurs particularly in
ruminants
Chain Elongation
 In eukaryotes, elongation of fatty acids (beyond C16) occur both in
the mitochondria as well as endoplasmic reticulum, especially in the
latter known as the microsomal system
 Reactions similar to FAS except involve CoASH derivatives e.g.
condensation of palmitoyl CoA (hence palmitate → palmitoyl CoA)
and malonyl CoA
Unsaturation of Fatty Acids
 Involves microsomal system called fatty acyl-CoA desaturase
 Palmitoyl CoA + NADH + H+ + O2 → Palmitoeyl CoA + NAD+ +
2H2O
 Mammals cannot introduce double bonds beyond ∆9 in fatty acids
 Unsaturated fatty acids like linoleic acid (ω6, 18:2, ∆9,12) and linolenic
acid (ω3, 18:3, ∆9,12,15), are the only two known essential fatty acids in
many animals, including humans; provided in the diet. 13
 Comparison of Fatty Acid Synthesis and
Degradation

Reduction
Dehydrogenation
(NADPH)
(FAD)

Hydration Dehydration

Reduction
Dehydrogenation
(NADPH)
(NAD+)

Condensa-
Thiolytic
tion
cleavage

Topic 8 : Metab. Liver 2 14

 Comparison of Fatty Acid Synthesis and
Degradation
Oxidative degradation Synthesis

Dehydrogenation Reduction
(FAD) (NADPH)

Hydration Dehydration
L-form D-form
Dehydrogenation Reduction
(NAD+) (NADPH)

Thiolytic Condensa-
cleavage tion
15
 Comparison of Fatty Acid Synthesis and
Degradation
Oxidative degradation Synthesis

Dehydrogenation Reduction
(FAD) (NADPH)

Hydration Dehydration
L-form D-form
Dehydrogenation Reduction
(NAD+) (NADPH)

Thiolytic Condensa-
cleavage tion
16
 Synthesis of Triacylglycerols
Glucose → → Dihydroxyacetone phosphate (1)
(Liver and adipose tissue)
Glycerol (liver) (2)

 Fatty acids are converted to

triacylglycerols (TAG) for storage
especially in adipose tissue and, lesser
extent, liver
 Stored TAG in dynamic state –
continuously broken down and
synthesized
 Phosphatidic acid is key intermediate not
only for TAG synthesis but also other lipids
e.g. phospholipids
(1) = Glycerol 3-phosphate dehydrogenase
(2) = Glycerol kinase (present in liver)
 Ketogenesis – Formation of Ketone Bodies
TCA Cycle Acetyl CoA Fatty acid

Ketogenesis 1

 Occurs when acetyl CoA accumulates

beyond cell’s capacity to oxidise it or to
synthesize fatty acids 2
CH3CO~SCoA
CoASH
 Occurs in the mitochondria, primarily in the
-
OOC-CH2
liver
 Extremely important during starvation
during which the brain adapts to using ketone 3

bodies for fuel

1 β-Ketothiolase
(Minor)
2 HMG-CoA synthase 4

3 HMG-CoA lyase
4 Β-Hydroxybutyrate dehydrogenase
Ketogenesis – Formation of Ketone Bodies
 Utilisation of Ketone Bodies
 Ketone bodies transported
from liver to other tissues and
used for energy production.
 Liver cannot convert
acetoacetate to acetoacetyl
CoA and hence cannot use
former for fuel
 Extrahepatic tissues including brain but not RBC (no mitochondria) efficiently
oxidise acetoacetate and hydroxybutyrate for energy production
 During starvation, glucagon secreted
 Adipose tissue, mobilisation of TAG → fatty acids
 Liver, increase breakdown of fatty acids → acetyl CoA
 Gluconeogenesis stimulated (Pyruvate → OAA →→ Glucose)
 Increase in acetyl CoA but low OAA, acetyl CoA accumulates
→ ketogenesis Topic 8 : Metab. Liver 2 20
Metabolism of Ketone Bodies

Topic 8 : Metab. Liver 2 21