LIPID

METABOLISM
MRS R. MAUTSA
• Lipid metabolism is the synthesis and degradation of lipids in
cells

• It involves
1. the breakdown and storage of fats for energy and
2. the synthesis of structural and functional lipids
 SOURCES OF FATTY ACIDS

1. Diet

2. Adipolysis – digestion of fats

3. De novo synthesis
 It is the synthesis of saturated fatty acids from acetyl CoA that is primarily
derived from glucose

 When an excess of dietary carbohydrate is consumed, glucose is converted to

acetyl CoA, which provides the two-carbon units that condense in a series of
reactions on the fatty acid synthase complex, producing palmitate.
 SITES FOR DE NOVO SYNTHESIS OF FATTY ACIDS

Occurs in the cytosol of the

• Liver
• Kidney
• Adipose tissue
• Lactating mammary gland.
 DE NOVO SYNTHESIS OF FATTY ACIDS

• Enzymes are in the cytosolic fraction of the cell

• Requires malonyl-CoA and acetyl-CoA

• acetyl CoA is found in the mitochondria

• Free palmitate is the product

- Palmitate is then converted to other fatty acids
DE NOVO SYNTHESIS OF FATTY ACIDS
 DE NOVO SYNTHESIS OF FATTY ACIDS

1. Acetyl-CoA is the starting material for FA synthesis.

- However, most Acetyl-CoA found in mitochondria

2. Acetyl-CoA must be transferred from the mitochondria to the

cytosol

3. BUT mitochondria not permeable to Acetyl-CoA

 TRANSPORTATION OF ACETYL CoA

1. Acetyl CoA is shuttled out of mitochondria as citrate

2. The mitochondrial inner membrane is impermeable to acetyl-CoA
3. Intra-mitochondrial acetyl-CoA first reacts with oxaloacetate to form
citrate (catalyzed by citrate synthase)
4. Citrate then passes into the cytosol through the mitochondrial inner
membrane on the citrate transporter.
5. In the cytosol, citrate is cleaved by citrate lyase regenerating acetyl-CoA.
TRANSPORTATION OF ACETYL CoA
 ENZYMES AND COFACTORS INVOLVED IN FATTY ACIDS
SYNTHESIS
Two main enzymes -
1. Acetyl CoA carboxylase
2. Fatty acid Synthase

Both the enzymes are multi-enzyme complexes

Coenzymes and cofactors are -

1. Biotin, NADPH, Mn²⁺, Mg²⁺
 FATTY ACID
SYNTHASE COMPLEX Acetyl transacylase AT

• A multi-functional enzyme is used to Malonyl transacylase MT

synthesize fatty acids
3-ketoacyl synthase KS

• Contains several enzyme activities

3-ketoacyl reductase KR
and an acyl carrier protein (ACP)
segment Enoyl reductase ER

• The fatty acid synthase complex is Thioesterase

located in the cytosol and, therefore,
it uses cytosolic acetyl CoA.
 FATTY ACID SYNTHASE COMPLEX

Fatty acid synthase

sequentially adds two-
carbon units from malonyl
CoA to the growing fatty
acyl chain to form
palmitate.
 FORMATION OF MALONYL CoA
1. Acetyl CoA is carboxylated to malonyl CoA by the enzyme
acetyl CoA carboxylase. (bicarbonate is the source of
CO2)

2. This is an ATP-dependent reaction & requires biotin for

CO2 fixation.
 CONVERSION OF ACETYL CoA TO MALONYL CoA
The overall reaction, which is spontaneous, is summarized as:
HCO3- + ATP + Acetyl-CoA  ADP + Pi + Malonyl-CoA
O

H3C C SCoA
acetyl-CoA
Acetyl-CoA
ATP + HCO3- Carboxylase
(inhibited by
ADP + Pi AMP-Activated
Kinase)
O
-
OOC CH2 C SCoA
malonyl-CoA
 ASSEMBLY OF A LONG CHAIN FATTY ACID
1. Once malonyl-CoA is synthesized, long carbon FA chains may be
assembled in a repeating four-step sequence

1. The growing fatty acid chain, attached to the fatty acid synthase
complex in the cytosol, is elongated by the sequential addition of
two-carbon units provided by malonyl CoA
2. The processes involved are-
i. Condensation
ii. Reduction
iii. Dehydration
iv. Reduction
These steps are repeated till a fatty acidwith 16 carbon atoms is synthesized
REPETITION OF THESE FOUR STEPS LEADS TO FATTY
ACIDS SYNTHESIS
 THE RESULT OF FATTY ACYL SYNTHASE
ACTIVITY
1. Seven cycles of condensation and reduction produce the 16-carbon
saturated palmitoyl group, still bound to ACP (on the fatty acid
synthase complex).

2. Chain elongation usually stops at this point, and free palmitate is

released from the ACP molecule by hydrolytic activity in the synthase
complex.

3. Smaller amounts of longer fatty acids such as stearate (18:0) are

also formed
 THE OVERALL REACTIONS FOR THE
SYNTHESIS OF PALMITATE FROM ACETY-
CoA CAN DIVIDED IN TWO PARTS
First, the formation of seven malonyl-CoA molecules:
7Acetyl-CoA + 7CO 2 + 7ATP 7malonyl CoA + 7ADP + 7Pi

Then the seven cycles of condensation and reduction

Acetyl-CoA + 7malonyl-CoA + 14NADPH + 14H⁺ palmitate + 7CO₂ + 8CoA + 14NADP⁺ + 6H₂O
The biosynthesis of FAs requires acetyl-CoA and the input of energy in the form of ATP and reducing power of NADPH.

The overall reactions

8Acetyl-CoA + 7ATP + 14NADPH + 14H ⁺ palmitate + 7CO₂ + 8CoA + 14NADP⁺ + 7ADP + 7Pi + 6H2O
BREAKDOWN OF FATTY ACIDS

OR

FATTY ACID OXIDATION

OVERVIEW

• The fatty acid chain is shortened by a recurring sequence of 4

reactions

• 4 enzyme catalysed reactions make up the 1st round of fatty acid

oxidation

• The FA chain is shortened by 2 carbon atoms as a result of these

reactions. FADH₂, NADH and acetyl CoA are generated

• Called Beta-Oxidation because most of the chemistry occurs on

the beta-carbon (beta to the carbonyl) per turn of the cycle
 OXIDATION OF FATTY ACIDS

1. Oxidation is the process where energy is produced by the

degradation of fatty acids.
2. There are several types of fatty acids oxidation.
i. β- oxidation of fatty acid
ii. α- oxidation of fatty acids
iii. ω- oxidation of fatty acids
 ß-OXIDATION OF FATTY ACIDS
1. Beta-oxidation is the process by which FAs, in the form of Acyl-CoA
molecules, are broken down in mitochondria and/or in peroxisomes
to generate Acetyl-CoA – which then enters the Krebs cycle

It occurs in many tissues including liver, kidney and heart but not the brain.
 STAGES
1. The beta oxidation of fatty acids involve three stages:

i. Activation of fatty acids in the cytosol

ii. Transport of activated fatty acids into mitochondria (carnitine

shuttle)

iii. Beta oxidation in the mitochondrial matrix

 1) ACTIVATION OF FA
FAs are degraded to acetyl CoA
1. FAs enter tissue cells in need of energy.
2. FAs must pass through the mitochondrial membrane to be
oxidized and to produce energy, but they cannot pass without
being activated first.
- FAs must be activated before they are degraded to produce energy. FAs are
activated in the cytosol, but oxidation occurs in the mitochondria
1) ACTIVATION OF FA
• This proceeds by acyl CoA synthetase present in cytosol
•Acyl CoA synthetase requires ATP, CoA-SH, Mg²⁺.
•The product of this reaction is F A acyl CoA and water.

A CoA molecule is added to the fatty

HS – CoA acid to produce acyl-CoA, converting
ATP to AMP in the process. Note
that in this step, the ATP is
converted to AMP, not ADP.
Thus, activation uses the equivalent
of 2 ATP molecules
 2)TRANSPORT OF FATTY ACYL CoA FROM CYTOSOL
INTO MITOCHONDRIA
Long-chain fatty acyl CoA moves across the inner mitochondria membrane with a special
transport mechanism called Carnitine shuttle.
The cytosol

The matrix
 2)TRANSPORT OF ACYL CoA INTO THE MITOCHONDRIA (RATE-
LIMITING STEP)
• Fatty acyl group from fatty acyl CoA is transferred to
carnitine to form acyl carnitine catalyzed by carnitine
acyltransferase I (CAT I), in the outer mitochondrial
membrane.
CAT I

• Acyl carnitine is then shuttled across the inner

mitochondrial membrane by a translocase enzyme.

• The acyl group is transferred back to CoA in the matrix by

CAT II
carnitine acyl transferase II.

• Finally, carnitine is returned to the cytosolic side by

translocase, in exchange for incoming acyl carnitine.
3) ß – OXIDATION IN THE MITOCHONDRIAL MATRIX

Step I – Oxidation by FAD linked dehydrogenase

Step II – Hydration by Hydratase

Step III – Oxidation by NAD linked dehydrogenase

Step IV – Thiolytic clevage Thiolase

 Oxidation

SUMMARY OF
REACTIONS Hydration

Oxidation

Thiolytic cleavage (thiolase)

 -OXIDATION REACTIONS FOR PALMITATE (C16)
Palmitoyl-CoA + 7 CoA + 7 FAD + 7 NAD+ + 7 H2O --> 8 acetyl CoA + 7 FADH2 + 7 NADH + 7 H+
ENERGETICS
3 ATP molecules per NADH 7 NADH × 3 ATP =21 ATP
2 ATP molecules per FADH2 7 FADH2 × 2 ATP = 8 ATP
12 ATPs per Acetyl-CoA 8 Acetyl-CoA x 12 ATP = 96 ATP
TOTAL COUNT = 131 ATP.

Activation energy loss – 2 ATP

Net energy output - 131 – 2
= 129 ATP
Regulation of Beta oxidation

• There are 2 types of regulation

1.Enzymatic regulation:- CAT-1 is the principal enzyme as it catalyzes
the rate-limiting step of beta-oxidation; which is the formation of
acylcarnitine. CAT-1 is allosterically inhibited by malonyl-CoA.

2.Hormonal regulation: It has 2 conditions

1. during well-fed conditions, the level of insulin increases which in turn
increases the level of malonyl-CoA. and this inhibits the beta-oxidation
process
2. during starvation, the level of glucagon increases, which decreases the level
of malonyl-CoA and stimulates beta-oxidation.
 IMPORTANCE OF MALONYL CoA
• Involved in the regulation of beta-oxidation of FAs
• When fuel is high, acetyl CoA is plentiful, and malonyl CoA is produced
• High levels of malonyl CoA inhibit acylcarnitine transferase/CPT1
(Enzyme involved in FA oxidation)
• No fatty acids can get into mitochondria for oxidation
• The level of malonyl-CoA thus determines the rate of fatty acid
oxidation
• Since malonyl CoA inhibits carnitine acyltransferase, a high rate of fatty
acid synthesis results in a low rate of fatty acid oxidation, and vice
versa