Fatty acid metabolism

Fatty Acid synthesis

• Lipids

• principal form of stored energy in most higher organisms,

• major constituents of membranes.

• Anabolism is not simply the reverse of catabolism.

• Lipid biosynthesis: endergonic and reductive.

• Regulated Pathway

• Location : Cytosol

• Starting substrate : Acetyl Co A (2C) → Malonyl CoA (3C)

• Electron Donor: NADPH

Subcellular localization of lipid metabolism
Transfer of acetyl groups from mitochondria to the cytosol.
Source of NADPH for fatty acid synthesis
Malonyl CoA

• formed from acetyl-CoA and bicarbonate

• Irreversible three step process

• Enzyme: acetyl-CoA carboxylase ( in cytoplasm)

• Prosthetic group: Biotin (Vit. H)

• covalently attached to the ε-amino group of a Lys residue of ACC.

• ATP dependent pathway

• Carboxyl group donor: bicarbonate (HCO₃⁻)

Malonyl CoA Synthesis
Malonyl CoA Synthesis
Fatty Acid synthesis

• repeating four-step sequence

• Enzyme: fatty acid synthase.

• Each cycle extends the fatty acyl chain by two carbons, using a saturated acyl group and an
activated malonyl group.

• The four-step process includes:

1. Condensation

2. Reduction

3. Dehydration

4. Reduction (converts the C-3 carbonyl to a methylene group)

Fatty Acid synthesis
Fatty Acid synthesis
Palmitate (16:0) synthesis
Fatty Acid synthase

• In mammals, the fatty acid synthase I (FAS I)

• seven active sites

• located in separate domains of a single multifunctional polypeptide (molecular

weight ~240,000), with two polypeptides forming a homodimer (molecular weight
~480,000).

• FAS I produces a single product through the cycle, with the acyl group covalently
linked to an acyl carrier protein (ACP).

• Acyl carrier protein (ACP)

• part of the FAS I polypeptide,

• ensuring that no intermediates are released during synthesis.

Fatty Acid synthase

• β-ketoacyl-ACP synthase (KS),

• malonyl/acetyl-CoA–ACP transferase (MAT),
• β-hydroxy-acyl ACP dehydratase (DH),
• enoyl-ACP reductase (ER),
• β-ketoacyl-ACP reductase (KR).
• ACP : acyl carrier protein.
• Thioesterase (TE) that releases the palmitate product from ACP
Fatty Acid synthesis
Fatty Acid synthesis Activated acetyl and malonyl
groups to form acetoacetyl-ACP
Fatty Acid synthesis
Stoichiometry
7 Acetyl-CoA + 7CO2 + 7ATP → 7 malonyl-CoA + 7ADP + 7Pi

Acetyl-CoA + 7 malonyl-CoA + 14NADPH + 14H+ → palmitate + 7CO2 + 8CoA + 14NADP+ + 6H2O

Notice that only six net water molecules are produced, because one is used to hydrolyze the
thioester linking the palmitate product to the enzyme

8 Acetyl-CoA + 7ATP + 14NADPH + 14H+ →palmitate + 8CoA + 7ADP + 7Pi + 14NADP+ + 6H2O
Regulation of fatty acid synthesis -regulation of Acetyl-CoA carboxylase-

-negative feed-back inhibition by

palmitate

- allosteric stimulation by citrate

-regulation by
covalent modification

Dephosphorylated form – active

-Polymerizes into
long filaments
insulin

Phosphorylation inactivates the

enzyme
Active dephosphorylated ACC
Routes of synthesis of unsaturated fatty acids
Routes of synthesis of unsaturated fatty acids
Triacylglycerols
 Triacylglycerols
• Major energy reserve
• Oxidation: 9 kcal/g (for carbohydrates: 4 kcal/g)
• 11 kg of 70 kg total body weight
• Site of accumulation: cytoplasm of ADIPOSE CELLS
• Liver converts excess carbohydrates to fats.
• About 40% of daily human energy comes from dietary triacylglycerols.
• Stored fat: starvation, hibernating animals, migrating birds, germinating seeds.
Processing of dietary lipids in vertebrates.

1. Ingestion of Triacylglycerols
2. Solubilization by Bile Salts
3. Lipase Action
4. Absorption into Intestinal
Mucosa
5. Reconversion to
Triacylglycerols
6. Formation of Chylomicrons
7. Transport via Lymphatic
System
8. Exchange of Apolipoproteins
9. Hydrolysis by Lipoprotein
Lipase
10. Uptake by Target Tissues
11. Utilization in Muscle and
Adipose Tissue
Mobilization of triacylglycerols stored in adipose tissue
1. Glucagon triggers cAMP production via
adenylyl cyclase.
2. PKA activates hormone-sensitive lipase
(HSL) and perilipin by phosphorylation.
3. CGI-58 recruits adipose triacylglycerol
lipase (ATGL) to break down
triacylglycerols to diacylglycerols.
4. HSL converts diacylglycerols to
monoacylglycerols.
5. Monoacylglycerol lipase (MGL)
hydrolyzes monoacylglycerols.
6. Fatty acids are released and transported by
serum albumin.
7. Fatty acids enter myocytes through specific
transporters.
8. In myocytes, fatty acids are oxidized to CO₂,
producing ATP for energy.
Fatty Acid Breakdown
Site of β-oxidation: mitochondria
Activation of fatty acids:
Fatty acid + CoA + ATP ⇌
fatty acyl–CoA + AMP + 2Pi
ΔG′° =−34 kJ/mol
Fatty acid entry into mitochondria
Fatty acid synthesis –Lipogenesis- not a reversal of degradation

Synthesis Breakdown
Site Cytosol Mitochondrial matrix

Intermediates bound to Acyl-carrier protein CoA

Enzymes Joined in a single Not associated

polypeptide chain (fatty acid
synthase)
Reducing equivalents NADPH NAD, FAD
Units Malonyl-CoA Acetyl-CoA
3-hydroxyacyl-derivative D-enantiomer L-enantiomer
Fatty Acid Catabolism
β-oxidation of fatty acids
Step 1: Dehydrogenation of Fatty Acyl-CoA

• Fatty acyl-CoA undergoes dehydrogenation,

• forming a double bond between the α and β carbons (C-2 and C-3).
• Produces trans-Δ²-enoyl-CoA (double bond in trans configuration).
Step 2: Hydration:
• Addition of Water to trans-Δ²-enoyl-CoA
• Enzyme: enoyl-CoA hydratase
• Product: L-β-hydroxyacyl-CoA.
Step3 : Dehydrogenation:
• L-β-hydroxyacyl-CoA is dehydrogenated
• Enzyme: β-hydroxyacyl-CoA dehydrogenase
• Product: β-ketoacyl-CoA.
• electron acceptor: NAD+ + e- →NADH
β-oxidation of Palmitate

Palmitoyl-CoA + 7CoA + 7FAD + 7NAD+ + 7H2O → 8 acetyl-CoA + 7FADH2 + 7NADH + 7H+

Palmitoyl-CoA + 7CoA + 7O2 + 28Pi + 28ADP → 8 acetyl-CoA + 28ATP + 7H2O

8 Acetyl-CoA + 16O2 + 80Pi + 80ADP → 8CoA + 80ATP + 16CO2 + 16H2O

Palmitoyl-CoA + 23O2 + 108Pi + 108ADP → CoA + 108ATP + 16CO2 + 23H2O

No. Of ATP = 7 x n +8 ( where n = no. of carbon )

ATP calculation in β-oxidation

No. Of ATP = 7 x n +8 ( where n = no. of carbon )

β-oxidation of MUFA
β-oxidation of PUFA
Regulation of fatty acid oxidation
Regulation of fatty acid oxidation
High energy state

hormones
(adrenaline, glucagon) Malonyl-CoA NADH Acetyl-CoA

Inhibition of carnitine Inhibition of thiolase

Inhibiton of Perilipin 3-hydroxyacyl CoA
acyltransferase I
Activation of hormone-sensitive lipase dehydrogenase

Inhibition of ß-oxidation
Entry of fatty Oxidation
Increased level of free fatty acids
acids into
mitochondria is
Oxidation inhibited
Formation of ketone bodies
• In humans and mammals, acetyl-CoA formed in the
liver from fatty acid oxidation can:

1. Enter the citric acid cycle.

2. Convert to ketone bodies: acetone,

acetoacetate, and D-β-hydroxybutyrate.

• Ketone bodies

• soluble compounds, not particulate,

• transported in blood and urine to other tissues.

Formation of ketone bodies
Ketone bodies as fuels
• D-β-Hydroxybutyrate, synthesized in
the liver,

• passes into the blood and thus to

other tissues,

• where it is converted in three steps to

acetyl-CoA.

• It is first oxidized to acetoacetate,

• which is activated with coenzyme A

donated from succinylCoA, then split
by thiolase.

• The acetyl-CoA thus formed enters

the citric acid cycle.