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Fatty Acid Degradation and Biosynthesis

Department of Biochemistry

Jeerus Sucharitakul
Fatty acid breakdown

Oxidation of hydrocarbon chain in fatty acid
•Free energy is stored in forms of FADH and NADH
•The final product from oxidation process is acetyl CoA.
•Oxidation occurs in matrix of mitochondria

Processes in fatty acid oxidation
I Activation of fatty acid (cytosol)
II Transport of activated fatty acid from cytosol into
intermembrane space and matrix
III Oxidation of hydrocarbon chains (matrix of mitochondria)
Activation of fatty acids
•Fatty acid is activated by forming a thioester link with CoA
•The reaction is catalyzed by acyl CoA synthase or fatty acid
thiokinase which is presented on the outer membrane of
mitochondria.
•Free energy for driving the reaction comes from hydrolysis
of pyophosphate (PPi) of ATP.
O O
R C O- + ATP + CoA R C S CoA + AMP + Pi2

∆Go = -15 kJ/mol

Pi2 + H2O Pi + Pi

∆Go = -19 kJ/mol
Activation of fatty acids
Transport of fatty acid into mitochondria

•Small and medium chains (≤10 C) acyl CoA; free diffusion
•Longer chains; enzymatic reaction, carnitine shuttle

CH3
H H2
H3C N+ CH2 C C COO- Carnitine
CH3 OH

•Facilitated diffusion by integral membrane protein
(carnitine transporter)
•Carnitine carries acyl CoA by covalent linkage to acyl group.
Transport of fatty acid into mitochondria

Carnitine transferase I; outer membrane of mitochondria
Carnitine transferase II; inner membrane of mitochondria
Carnitine-mediated entry process is the rate-limiting step.
Oxidation of fatty acids

•Cleavage bond of hydrocarbon chain at β-C (β-oxidation)
by two carbon producing of acetyl CoA.
•Hydrocarbon is an inert compound therefore, it is activated
by reduction forming of double bonds.

β-oxidation

Acetyl CoA FADH2 NADH

Oxidative phosphorylation
Oxidation of fatty acids

I Oxidation III Oxidation

IV Thiolysis
II Hydration
Oxidation of fatty acids

β -oxidation of saturated
fatty acids

β -oxidation of unsaturated
fatty acids
-Most lipid in animals and plants
Are unsaturated fatty acids
-Double bonds are in cis
conformation.
β -oxidation of unsaturated fatty acids
H2 H2 H2
O Monosaturated fatty acids
C C C C C C SCoA
H H
•Double bond at C-3 prevents
β-oxidation reduction the bond between
C-2 and C-3
O
H2
C C C C SCoA •Enzyme enoyl-CoA hydratase
H H
cis-∆3-enoyl CoA
is only specific for substrate-
trans conformation.
Isomerase
•Enzyme isomerase translocate
H O double bond from cis to trans
H2
C C C C SCoA conformation without reduction.
H
trans-∆2-enoyl CoA
β -oxidation of monounsaturated fatty acids
β -oxidation of polyunsaturated fatty acids
β -oxidation of polyunsaturated fatty acids
β -oxidation of fatty acids

Even-number fatty acids Odd-number fatty acids

β-oxidation
C3
Acetyl CoA Acetyl CoA + Propionyl CoA

Carboxylation

Succinyl CoA
TCA cycle
Conversion of propionyl-CoA into succinyl-CoA
B12 is a cofactor of methylmalonyl-CoA mutase.
Vitamin B12 deficiency
•Serious disease; pernicious anemia
•Vit B12 can not be synthesized by animals or plants
but only few microorganism can do it.
•Vit B12 is synthesized by intestinal bacteria
•Absorption of Vit B12 in intestinal lumen requires a
glycoprotein, intrinsic factor.

Pernicious anemia
-Genetic deficiency disease, for intrinsic factor
-Low production of hemogolbin
-Low production of erythrocytes
-Severe impairment of central nervous system
Animals can not use acetyl CoA for gluconeogenesis.

•Animals can not convert acetyl CoA
into pyruvate or oxaloacetate.

•No net conversion of carbons from
acetyl CoA produced from fatty acid
oxidation into oxaloacetate.

•Plants have the pathway for conversion
of acetyl CoA into oxaloacetate for
glucose synthesis and for other
precursors, glyoxylate cycle.
Ketone bodies
•Excess production of Acetyl-CoA from β-oxidation
can be converted into ketone bodies in the liver.
•Ketone bodies can be used as a fuel by other tissues
(extrahepatic tissues) such as brain, heart, renal cortex
and muscle under starvation.
Formation of ketone bodies from acetyl Co-A
Degradation of β -hydroxybutyrate as a fuel by
extrahepatic tissues
Liver acts as a producer of
β-hydroxybutyrate but can
not be a consumer because
it lacks of enzyme thiolase.
Conditions for ketone bodies production

I Starvation; deplete of glucose, glycogen and intermediates in
TCA cycle for gluconeogenesis
-At this condition all tissues divert to use fat as a energy
source.
-Liver produces ketone bodies for extrahepatic tissues

II Untreated diabetes; lack of insulin
-Extrahepatic tissues can not take up glucose efficiently from
Blood.
-lack of intermediate in TCA cycle due to high rate of
gluconeogensis.
-Accumulation of acetyl-CoA
-Characteristic odor to the breath from volatile acetone
Acidosis
-Lowered blood pH from β-hydroxybutyrate

-Coma and death

-High level of ketone bodies in blood and urine, ketosis
-Collectively called ketoacidosis

-People during very low-calorie diets, body uses fat from
Adipose tissue as a major source of energy are risky to
ketosis.
Lipid biosynthesis

Lipids have many various functional roles in cells.

•Composition of cell membranes
•Pigments; retinal, carotene
•Cofactor; vitamin K
•Bile salts
•Hormones
•Intracellular mediators
Lipid biosynthesis
•Fatty acid synthesis occurs in different pathways and
different compartment from fatty acid breakdown.
•In most higher eukarytoes, fatty acid biosynthesis occurs in
cytosol.
•The first intermediate for lipid biosynthesis is three-carbon
compound, malonyl-CoA.
•Formation of malonyl-CoA is a rate-limiting step in this process.
•Malonyl-CoA is synthesized by acetyl-CoA carboxylase.

O O
H2
C C C
-
O S CoA
Malonyl-CoA
Lipid biosynthesis

Acetyl-CoA carboxylase
•Multifunctional enzyme
•Coenzyme; biotin (CO2 carrier)
Lipid biosynthesis

Enzyme catalyzing fatty acid synthesis is fatty acid synthase.

Substrates for fatty acid synthase
-Malonyl-CoA
-NADPH
-Acetyl-CoA
-ATP

Palmitate (saturated C16) is the principle product of the fatty
acid synthase system in animal cells.
Fatty acid synthase
•Multifunctional enzyme
•The product from chain elongation is held by
ACP domain (acyl carrier protein)
•Enzyme is located on cytosolic
surface of SER.
Fatty acid synthase
Channeling of substrates for fatty acid synthesis

Pentose phosphate pathways Carbohydrates, amino acids,
(cytosol) β-oxidation (mitochondria)

NADPH Acetyl-CoA

Fatty acid synthesis

Reoxidation of malate
to pyruvate by malate
enzyme
Malonyl CoA
Source of NADPH in cytosol
Cytosolic NADPH is generated by (hepatocyte and adipocyte)

I Pentose phosphate pathway; animals
II Malic enzyme; plants

2NADP+ 2NADPH + H+

Glucose 6-phosphate Ribulose 5-phosphate

COO- NADP+ NADPH + H+ COO-
HCOH
C O + CO2
CH2
Malic enzyme COO-
COO-

Malate Pyruvate
Acetyl CoA shuttle
•Acetyl CoA used in fatty acid synthesis is formed in
matrix of mitochondria.
•Inner membrane of mitochondria is impermeable to
acetyl CoA.
•Acetyl CoA is passed into cytosol in form of citrate.
•Citrate is hydrolyzed by enzyme citrate lyase releasing
acetyl-CoA in cytosol

Citrate synthase
Oxaloacetate + Acetyl-CoA Citrate + CoA-SH

Citrate lyase
Citrate + CoA-SH + ATP Oxaloacetate + Acetyl-CoA + ADP + Pi
Acetyl CoA shuttle
Regulation of fatty acid synthesis

Citrate

Citrate lylase Insulin

Acetyl-CoA

Acetyl-CoA Glucagon
carboxylase Epinephrine

Malonyl-CoA

Palmitoyl-CoA
Long chain fatty acid synthesis

Palmitate (C16) is the
principal product of the fatty
acid synthase system in
animal cells.

•Long chain fatty acids are
synthesized by other system, fatty
acid elongation system which is in
SER and mitochondria.

•The mechanism for elongation is
similar to that of fatty acid synthase.
Synthesis of unsaturated fatty acid (desaturation)

•Palmitate (C16) and stearate (C18) are precursors for
unsaturated fatty acids.
•Most common monounsaturated fatty acids of animal
tissues are palmitoleate, 16:1∆9 and oleate, 18:1 ∆9.

•The double bond is introduced by
oxidation, which is catalyzed by
enzyme fatty acyl-CoA desaturase
in ER.
•NADP+ is an final electron acceptor.
Synthesis of unsaturated fatty acid (desaturation)
•Only plants can synthesize
polyunsaturated fatty acids, linoleate
and linolenate.
•Both of those fatty acids are
essential fatty acids for mammals as
precursors for other polyunsaturated
fatty acids.
•Arachidonic acid (ecosanoid) is
an essential of regulatory lipids.
Ecosanoids (C20) are biological signaling in cells.

Arachidonic acid

Cycloxygenase Lipoxygenase

I Prostaglandins; Leukotrienes;
-pain, inflammation, fever, -Contraction of smooth
wake-sleep cycle muscle in lung
II Thromboxanes; -Allergic reaction or
-vasoconstriction hypersensitivity
-blood clotting
-platelet aggregration
Ecosanoids (C20) are biological signaling in cells.

Cyclic pathway of cycloxygenase
Ecosanoids (C20) are biological signaling in cells.
Linear pathway of lipooxygenase
Biosynthesis of triacylglycerols

•Synthesis of storage lipids
•Synthesis of membrane
phospholipids

•Glycerol 3-phosphate is a
precursors for biosynthesis of
triacylglycerols.
•Reactions is catalyzed by two
pathways using enzyme
glycerol 3-phosphate
dehydrogenase for reduction
and glycerol kinase.
Biosynthesis of triacylglycerols

Fatty acyl-CoA

Phosphatidic acid is an intermediate for triacylglycerol, and
for glycerophopholipids (components of cell membranes).
Biosynthesis of triacylglycerols

Phosphatidic acid is an intermediate for
triacylglycerol, and for glycerophopholipids
(components of cell membranes).
Hormonal regulation triacylglycerol synthesis
Triacylglycerol cycle
Biosynthesis of membrane phospholipids
Constructions of membrane phospholipids
I Synthesis of backbone molecule (glycerol or sphingosine)

II Attachment of fatty acids through
ester or amide

III Addition of head group through
phosphodiester or ester

IV Alteration of head group to the
desired product.
CDP (cytidine diphosphate) is used for action
either diacylglycerol or head group.
Regulations of fatty acid degradation and
biosynthesis
Biosynthesis of cholesterol and steroids

•27-carbon compound
•Component of cellular membranes
•Precursor for steroid hormones
•Precursor for bile acid
•One of risk factors for cardiovascular diseases

-Mammalian cells can synthesize cholesterol from simple
molecules, acetyl-CoA.
-The isoprene unit is an intermediate for cholesterol
synthesis.

CH3
H2C C CH CH2

Isoprene unit
Biosynthesis of cholesterol and steroids

Committed step for
cholesterol synthesis

Mevalonate is oxidized into C5 isoprene unit by decarboxylation.
HMG CoA reductase

•Committed step and regulation step for cholesterol
Synthesis
•Feedback inhibition by high level cholesterol
•Target for therapy of hypercholesterolemia, competitive
inhibition to decrease rate for cholesterol synthesis
-Lovastatin; fungal products
•Much of cholesterol synthesis in vertebrates takes place
in the liver.
Steroid hormones
Bile salts
•Bile acids are polar derivatives of cholesterol with amino acid.
•Synthesis from liver in form of glycolate and taurocholate.
•Bile salts are stored in and concentrated in the gall bladder,
and released into intestine for lipid digestion.
O O
OH -
O OH
N SO3-
N
H O H

HO OH HO OH

Glycolate Taurocholate

Amide linkage with glycine Amide linkage with taurine
Vitamin D
•Vitamin D (cholecalciferol)
•Synthesis by skin from 7-dehydrocholesterol in a
photochemical reaction
•Regulation of gene expression

7-dehydrocholesterol
UV light

Cholecalciferol (Vitamin D3)

Liver and Kidney

Dihydroxycholecalciferol
(reactive compound)