DENOVO SYNTHESIS OF FATTY

ACIDS
FA oxidation FA synthesis
• Starvation • Well fed state
• Mitochondria • Cytoplasm
• NADH & FAD • NADPH
• CoA • ACP
 Fatty Acid synthesis

• Not a simple reversal of the beta oxidation

• Different pathways
• Catalyzed by different sets of enzymes
• Take place in different parts of the cell
 Fatty Acid synthesis
Organs involved:

• Liver

• Lactating mammary gland

• Adipose tissue

• Kidney, Brain, Lung

 Fatty Acid synthesis
• Acetyl co A from glucose - primary substrate
for lipogenesis

• Synthesized by sequential addition of all

carbon units to the activated carboxyl end of
a growing chain
 Fatty Acid synthesis
Co factor requirements:
• NADPH
• ATP
• Biotin
• HCO3-
 Fatty Acid synthesis
• Saturated, straight chain C16 acid- palmitic acid
- is first synthesized, and all other fatty acids
are made by modification of palmitic acid
Transfer of acetyl CoA to
the cytoplasm
 Sources of NADPH

• Malic enzyme
 Sources of NADPH

Pentose Phosphate Pathway

 Fatty Acid synthesis
Step 1 - Committed step
• The formation of malonyl-CoA from acetyl-
CoA
• Catalyzed by acetyl-CoA carboxylase
• Irreversible process
 Fatty Acid synthesis

Acetyl-CoA carboxylase
• A multi enzyme protein with variable
number of identical subunits
 Fatty Acid synthesis
Acetyl-CoA carboxylase – each subunit contain
1.Biotin
2.Biotin carboxylase
3.Biotin carboxyl carrier protein
4.Transcarboxylase
5.Regulatory allosteric site
 Fatty Acid synthesis
Reaction take place in two steps:

• Carboxylation of biotin involving ATP

• Transfer of carboxyl group to acetyl coA to

form malonyl coA
 Enzyme-biotin
-
HCO3 + ATP
1
ADP + Pi
-
Enzyme-biotin-CO2
O
ll 2
CH3-C-SCoA
Enzyme-biotin
acetyl-CoA
O
-
ll
O2C-CH2-C-SCoA
malonyl-CoA
 Fatty acid synthase complex

• Fatty acid synthase I (FAS I ): vertebrates and

fungi

• Fatty acid synthase II (FAS II): plants and bacteria.

 Fatty acid synthase complex

FAS I :
• Single multifunctional polypeptide chain
• Seven active sites for different reactions
• Lie in separate domains
• Functions as a homodimer
• The subunits function independently
 Fatty acid synthase complex

• Seven enzymes are organized into three

domains joined by flexible regions

 Fatty acid synthase complex
1st domain
• Acetyl transacylase (AT)
• Malonyl transacylase (MT)
• Condensing enzyme (CE) / Beta keto acyl
synthase
 Fatty acid synthase complex
2nd domain
• Acyl carrier protein (ACP)
• ß-ketoacyl reductase (KR)
• Dehydratase (DH) / Hydratase
• Enoyl reductase (ER)
 Fatty acid synthase complex

3rd domain
• Thioesterase (TE)
 Fatty acid synthesis

• The intermediates remain covalently attached

as thioesters to one of two thiol groups
 Fatty acid synthesis

1. SH group of Cys residue of CE /Beta keto acyl

synthase

2. SH group of Acyl carrier protein

 Acyl carrier protein (ACP)

• Prosthetic group 4'-phosphopantetheine

serves as a flexible arm

• Carries the reaction intermediates from one

enzyme active site to the next
Acyl carrier protein (ACP)

• Hydroxyl group of a
Ser residue in ACP
 Fatty acid synthesis

Charging of FAs

• Transfer of acetyl group of acetyl-CoA to the

Cys –SH group of the Condensing Enzyme (CE)

• Catalyzed by Acetyl transacylase

 Fatty acid synthesis
Charging of FAs
• Transfer of the malonyl group from malonyl-
CoA to the -SH group of ACP

• Catalyzed by Malonyl transacylase

 Fatty Acid synthesis
Condensation
• Condensation of acetyl and malonyl groups to
form 3 keto acyl enzyme

• A molecule of CO2 released

 Fatty Acid synthesis
Reduction
• Reduction of the carbonyl group at C-3 to
form 3 Hydroxyacyl enzyme
• Catalyzed by ß-ketoacyl reductase
• The electron donor - NADPH
 Fatty Acid synthesis
Dehydration
• Water removed from C-2 and C-3 to yield a
double bond 2,3 unsaturated acyl enzyme

• Catalyzed by Hydratase / Dehydratase

 Fatty Acid synthesis
Reduction
• Double bond of 2,3 unsaturated acyl enzyme
is reduced to form acyl / butyryl-enzyme

• By Enoyl reductase (ER)

• Electron donor - NADPH

 Fatty Acid synthesis
• Charging
• Condensation
• Reduction
• Dehydration
• Reduction
• One cycle produce four carbon, Butyryl
enzyme
 Fatty Acid synthesis

• Next cycle of four reactions lengthens the

chain by two more carbons
 Fatty Acid synthesis
Second cycle:
• The butyryl group transferred from ACP to the
Cys -SH group of CE

• Another malonyl group is linked to ACP

• Product of condensation six-carbon acyl group

 Fatty Acid synthesis
• The sequence of reactions is repeated six more
times until a saturated 16 carbon palmityl chain
has been assembled

• Chain elongation stops at this point

• Free palmitate released from the ACP

by thioesterase ( TE )
 Overall reaction

• 8 Acetyl-CoA+ 7 ATP + 14NADPH+ 14H

+

• Palmitate+ 8 CoA+7 ADP+ 7Pi+

14NADP+ 6H2O
 Energetics

• 3ATP /1 Acetyl CoA

• 2ATP for transport from mitochondria

Regulation of denovo synthesis of
fatty acids
 Regulation

• Fatty acid synthesis is maximal when

carbohydrates and energy are plentiful
 Regulation

• Acetyl-CoA carboxylase reaction

:The rate limiting step

• Important site of regulation.

 Regulation

Short term regulation

• Allosteric & Covalent modification of enzymes

Long term regulation

• Regulation of gene expression

 Regulation

• Allosteric:Citrate and palmitoyl CoA

• Covalent: Hormonal

• Gene expression:Diet /Insulin

 Allosteric
• Palmitoyl-CoA: Feedback inhibitor

• Citrate :Allosteric activator

 Covalent modification
Insulin Vs Glucogon,epinephrin
 Long term regulation

• By changing the gene expression of adaptive

enzymes
• Concentration of enzymes
 Increased during fed state
 Decreased in starvation, high fat diet & in
diabetes mellitus
 Long term regulation

• Insulin : Increase concentration of

enzymes
• Glucagon : Decrease the concentration of
enzymes
• Increase PUFA in diet : inhibition of
lipogenesis
 Fate of palmitic acid

• Formation of acylglycerols

• Chain elongation or desaturation

• Esterification to cholesterol ester

 Fatty acid Elongation

• Site : endoplasmic reticulum / mitochondria

• The elongation beyond 16-carbon length

• By the action of multienzyme complex - Fatty acid

elongase

• Requires another set of enzymes

 Fatty acid Elongation

• Occurs by the addition of 2-carbon fragments

derived from malonyl-CoA

• NADPH - reductant

• Coenzyme A is the acyl carrier in the reaction

 Fatty acid Elongation

• The mechanism similar to that in palmitate

synthesis

• Occur in mitochondria

• Donation of two carbons by malonyl-CoA

• Followed by reduction, dehydration, and reduction

 Fatty acid Elongation

• Very long chain (22-24 carbon) fatty acids are

produced in the brain

• Elongation in the brain increases rapidly during

myelination

• Required for the synthesis of sphingolipids

 Desaturation of fatty acids

• The body has a requirement for mono- and polyunsaturated

fatty acids, in addition to saturated fatty acids.

• Some are supplied in the diet

• Linoleic and linolenic : essential fatty acids

 Desaturation of fatty acids

• By an oxidative reaction catalyzed by fatty acyl-CoA

desaturase

• A mixed-function oxidase

• Site – Mainly smooth endoplasmic reticulum of

hepatocytes
 The Desaturation system

• Molecular oxygen

• NADH / NADPH

• Cytochrome b5

• Cytochrome b5 reductase
 Desaturation of fatty acids
• Palmitate and stearate are precursors of

2 most common un saturated fatty acids of animal

tissues

– Palmitoleate - 16 :1(▲9)

– Oleate - 18:1(▲9);
 Desaturation of fatty acids

• The first double bond introduced into a saturated

fatty acids is nearly always in the Δ9position

• Additional double bonds can be introduced at the

Δ4, Δ5, Δ6 positions
 Why essential fatty acids are essential?

• Double bonds can be introduced at the Δ4, Δ5, Δ6

and Δ9 positions in most mammals

• But cannot introduce additional double bonds

between C10 and the methyl-terminal end
 Why essential fatty acids are essential?

Mammals cannot synthesize

• Linoleic acid 18:2(▲9,12)

• α-linolenic acid 18:3(▲9,12,15)

 Why essential fatty acids are essential?

• Necessary precursors for the synthesis of other

products

• They must be obtained from dietary plant material

 Synthesis of polyunsaturated fatty acids

• Essential for maintenance of the physical state

of stored TAGs and membrane phospholipids

• Synthesized by humans through a combination

of elongation and desaturation reactions

• Site : microsomes
 Essential fatty acid deficiency

• Rare

• EFAs – high in vegetable oils

• Precursors for prostaglandin, thromboxane,

leukotriene & lipoxin formation

• Found in structural lipids of the cell

 Essential fatty acid deficiency

• Concerned with structural integrity of mitochondrial

membrane

• Arachidonic acid – 5-15% FAs in phospholipids

 Essential fatty acid deficiency

• DHA - w 3 (22:6) , synthesised from linolenic acid /

obtained from fish oils

• High amount in retina, cerebral cortex, testis &

sperm

• Needed for the development of brain and retina

 Essential fatty acid deficiency
Clinical features:
• Cystic fibrosis, Retinitis pigmentosa
• Scaly dermatitis, hyperkeratosis
• Acrodermatitis enteropathica
• Hepatorenal syndrome
• Multisystem neuronal degeneration
• Hypercholestrolemia
 Essential fatty acid deficiency

Prevention :

• Intake of essential fatty acids -1 – 2% of total caloric

requirement