Beta Oxidation of Fatty acids

Beta Oxidation is the process where energy is produced

from Lipids.

Beta oxidation is the oxidation over the β -Carbon of

hydrocarbon chain of fatty acids by a sequential cleavage of
two carbon atoms.
The break down of a fatty acid to acetyl-CoA
STRICTLY AEROBIC

Occurs in the mitochondria

Acetyl-CoA is fed directly into the Krebs cycle

Overproduction causes KETOSIS

→ β -oxidation takes place in Mitochondria
→ Fatty acids which are participating in β-oxidation
undergo activation to form Fatty acyl CoA
→ Activation of fatty acid takes place in cytoplasm,
requires 2 high energy bonds and enzyme is
Thiokinase or fatty acyl Co A synthetase.
→ The activated fatty acids which are present in
cytoplasm enters into mitochondria with the help of
Carnitine.
 CARNITINE is β – hydroxy trimethyl amonium butrate
It is synthesized from Lysine and Methionine in Liver &
Kidney.
Lipid Metabolism
 Transport into Mitochondria depends on Carnitine

+
N(CH3)3
FA~CoA Acyl transferase I
HS-CoA CH2
Carnitine
FA~Carnitine H-C-OH
CH2
Translocase
COO-
Carnitine
FA~Carnitine Carnitine
HS-CoA
Acyl transferase II
FA~CoA
β – oxidation proper

There are 4 steps in β – oxidation

Step I – Oxidation by FAD linked dehydrogenase

Step II – Hydration by Hydratase

Step III – Oxidation by NAD linked dehydrogenase

Step IV – Thiolytic clevage Thiolase

Beta Oxidation
Beta Oxidation
Beta Oxidation
Beta Oxidation
Beta Oxidation Palmitic (16 c)
 Beta Oxidation Palmatic acid(16 c)

8
7
Energetics:
Palmitic acid (16 C) needs 7 cycles of beta-
oxidation, which gives rise to
8 molecules of Acetyl Co A

8 acetyl Co A x 12 = 96 ATP
7 FADH2 x 2 = 14 ATP
7 NADH x 3 = 21 ATP
Total 131 ATP
In initial activation Palmic acid → Palmitoyl Co A
requires 2 high energy phosphates, so net is 131 – 2
= 129 ATP
Regulation:
(1) Availability of FFA regulates β-oxidation. The
availability of FFA is controlled by
Glucagon : Insulin ratio
Glucagon Increases FFA level
Insulin decreases FFA level

(2) CAT – I regulates entry of Fatty acyl Co A into

mitochondria.
Malonyl Co A Inhibits CAT – I activity.
 Biosynthesis of Fatty acids

The excess dietary Carbohydrates & Proteins can

be converted to fatty acids and are stored as Tri acyl
Glycerol.
Denovo synthesis of Fatty acids takes place in
Liver, Kidney, adipose tissue and Lactating
Mammary glands.
Site: Cytoplasm of the cell
Requirements:
Acetyl CoA – source of Carbon atoms
NADPH – provides reducing equivalents
ATP – energy
═ Fatty acid synthesis in 3 stages

(i) Production of Acetyl CoA & NADPH

(ii) Conversion of acetyl CoA to Malonyl CoA

(iii) Reactions of Fattyacid synthase complex.

Production of Acetyl CoA & NADPH
Acetyl CoA is produced in mitochondria from oxidation
of pyruvate
fatty acids
degradation of Amino acids
Ketone bodies

 Mitochondrial membrane is impermeable to Acetyl

CoA
Transfer of Acetyl CoA from mitochondria to cytoplasm
═ In mitochondria Acetyl CoA + Oxaloacetate
to form Citrate
═ Citrate is freely transferable
═ In cytoplasm Citrate is cleaved
to Acetyl CoA & Oxaloacetate
oxaloacetate
↓
Malate
NADPH Malic enzyme

Pyruvate
 Citrate As Carrier of Acetate Groups

Cytosol Mitochondria
Glucose Pyruvate Pyruvate Pyruvate Acetyl CoA
Dehydrogenase
Malic enzyme

Malate
Malate Oxalo-
dehydrogenase acetate
Citrate
Acetyl CoA
Oxaloacetate
ATP-Citrate
Lyase
Note: Acetyl CoA
Citrate
cannot be converted
to glucose
Mitochondrial
membrane 21
 Sources of NADPH

NADPH
↑
1) Malic enzyme (Malate → Pyruvate)

2) HMP Shunt
 Regulation of Fatty acid Synthesis
1) Acetyl CoA Carboxylase →Important step in FA Synthesis

Acetyl CoA → Malonyl CoA

This enzyme is activated by citrate
Inhibited by Palmitoyl CoA

This enzyme is also activated by Insulin

Inactivated by Glucagon, epinephrine & nor epinephrine.

Diet also influences fatty acid synthesis.

 Synthesis of TAG
(I) Stage :→ Synthesis of Glycerol – 3 – phosphate
a) In liver Glycerol is converted to Glycerol – 3 – phosphate
by Glycerol Kinase

Glycerol Glycerol – 3- phosphate

b) In liver & adipose tissue from Glycolysis

DHAP is converted to Glycerol -3 – phospate by
Glycerol – 3- Phosphate dehydrogenase

DHAP Glycerol – 3 – Phosphate

(II) Stage : Addition of acyl groups (fatty acids) to

Glycerol – 3 – Phosphate
Acyl transferases catalyes the transfer acyl groups
Glycerol – 3 – Phosphate
Fatty acyl CoA
CoA

Lysophosphatidic acid
Fatty acyl
CoA CoA

phosphatidic acid

P1 Phosphatase

Diacyl glycerol
Fatty acyl
CoA CoA

Triacyl glycerol
 Transamination
Salient features:
► Transfer of amino group from an amino acid to keto acid is
known as Transamination.

► It involves interconversion of a pair of α-amino acid & a pair

of α-keto acids and is catalyzed by transaminases or
aminotransferases.

► Pyridoxal phosphate is the coenzyme essential for

transaminase activity.

► Alanine-pyruvate transaminase or Alanine transaminase &

glutamate-α-ketoglutarate transaminase or glutamate
transaminase make a significant contribution & transfer amino
acids from Alanine or glutamate.

► Transamination is a reversible process.

► There is no free ammonia liberated ,only the transfer of
amino group occurs.

► Each transaminase is specific for specified pair of amino

acids and keto acids as one pair of substrates.

► Most amino acids are substrates for Transamination except

lysine, Threonine, Proline and Hydroxy Proline.

► δ-amino group of ornithine also undergoes Transamination.

► Transamination is important for distribution of amino group

and production of non-essential amino acids & involves both
catabolism and anabolism of amino acids.

► Transamination diverts excess amino acids towards energy

generation.
► Amino acids undergo Transamination to finally concentrate
nitrogen in glutamate.

► Glutamate is the only amino acid that undergoes oxidative

deamination to liberate ammonia for urea synthesis.

► Serum transaminases are important for diagnostic and

prognostic purposes.

Mechanism:
It involves two stages:

► Stage 1:

Transfer of amino group to coenzyme pyridoxal phosphate

to form pyridoxamine phosphate.
► Stage 2:

The amino group of pyridoxamine phosphate is transferred to

keto acid to produce new amino acid with pyridoxal phosphate is
regenerated.

All transaminases require pyridoxal phosphate (PLP), a

derivative of vitamin B6.

Aldehyde group of PLP is linked with ε-amino group of lysine

residue forming Schiff base.

When amino acid comes in contact with the enzyme it displaces

lysine and new Schiff base linkage formed by non covalent
forces.

Snell & Braustein proposed a Ping Pong Bi Bi mechanism involving

series of intermediates in Transamination reactions,
 Deamination
 Removal of amino group from amino acids as ammonia is
called Deamination

 liberation of ammonia is used for synthesis of urea cycle and

the carbon skeleton of amino acids is converted to keto
acids.

 Deamination my be either oxidative or non-oxidative.

 both transamination and Deamination involve in using

glutamate as central molecule.

 1.oxidative Deamination:

 it is the liberation of free ammonia from the amino group of

amino acids coupled with oxidation.
 it takes place mostly in liver and kidney

 the role of glutamate with enzyme glutamate dehydrogenase

(GDH) serves as collection centre for amino groups to liberate
ammonia.

 this enzyme is unique and it can use either NAD+ or NADP+

as coenzyme.

 conversion of L-Glutamate to α- Ketoglutarate occurs

through an intermediate product called α- iminoglutarate with
help of GDH.

 GDH reaction is important as it links up glutamate

metabolism with TCA Cycle through α- Ketoglutarate

 GDH is involved in both catabolic and anabolic reactions.

 GDH is a zinc containing mitochondrial enzyme.

 GDH is controlled by allosteric regulation.

 GTP, ATP, Steroids & Thyroid hormones inhibit GDH.

 GDP & ADP activate GDH.

 L-Amino acid oxidase & D- amino acid oxidase are

flavoproteins, possessing FMN & FAD.

 These act on amino acids to produce α-keto acids and

ammonia.

 oxygen is reduced to hydrogen peroxide and decomposed by

catalase.
 Activity of L- amino acid oxidase is low & D-amino acid
oxidase is high in liver and kidney.

 L-amino acid oxidase does not act on glycine and

dicarboxylic acids.

 if enzyme catalase is absent α-keto acid produced by

oxidative Deamination is dacarboxylated by hydrogen peroxide
forming carboxylic acid with one carbon less.

 the enzyme L-amino acid oxidase has no effect on glycine.

 L-amino acid oxidase does not have major role in catabolism

of mammalian amino acids and formation of ammonia.
2. Non-oxidative Deamination:

 Serine, Threonine & homoserine are hydroxyl amino acids

which undergo non-oxidative Deamination , catalyzed by PLP-
dependent dehydrases.

 Sulfur amino acids like Cysteine & Homocysteine undergo

Deamination coupled with desulfhydration to give keto acids.

 Urocunoate acts on histidine to liberate ammonia by non-

oxidative Deamination.

Production of Urea
Organisms can excrete excess N as
 Ammonia (ammonotelic; e.g., aquatic animals)
 Urea (ureotelic; terrestrial animals)
 Synthesized in liver by urea cycle (discovered by Hans Krebs,
before he elucidated the TCA)
 Uric acid (uricotelic: birds, reptiles, dinosaurs?)

H
N
NH
O

N
H N O
H
uric acid
 Overall Reaction for Urea Cycle
NH3+
H2
NH3 + HCO3- + -O2C C C CO2-
H

3 ATP

2ADP + 2Pi +AMP + PPi

-
H2N C NH2 + O2C C C CO2-
H H
The Steps of the Urea Cycle
1. Carbamoyl Phosphate Synthetase
I (CPS I; mitochondrial form)
2. Ornithine Transcarbamoylase:
transfer of carbamoyl group to O

ornithine HO

NH2
NH2

ornithine

3. Argininosuccinate synthetase
Mechanism of carbamoyl synthetase I
Acquisition of 1st N

Mechanism of argininosuccinate
Acquisition of 2nd N