AMINO ACID METABOLISM

 General reactions of amino acid metabolism: Transamination,

deamination & decarboxylation
 Urea cycle and its disorders

 Catabolism of phenylalanine and tyrosine and their metabolic

disorders (Phenylketonuria, Albinism, alkeptonuria, tyrosinemia)
 Synthesis and significance of biological substances; 5-HT,
melatonin, dopamine, noradrenaline, adrenaline
 Catabolism of heme; hyperbilirubinemia and jaundice
 Metabolism of amino acids

• Proteins are the most abundant organic compounds and constitute a major
part of the body dry weight (10-12 kg in adults).

• Proteins are nitrogen-containing macromolecules consisting of L-α-amino

acids as the repeating units.

• Of the 20 amino acids found in proteins, half can be synthesized by the body
(non-essential) while the rest have to be provided in the diet (essential amino
acids).

• The proteins on degradation (proteolysis) release individual amino acids.

Amino acids are not just the structural components of proteins.

• Each one of the 20 naturally occurring amino acids undergoes its own
metabolism and performs specific functions.
 Metabolism of amino acids

• Some of the amino acids also serve as precursors for the synthesis of
many biologically important compounds

e.g. melanin, serotonin, creatine etc.

• Certain amino acids may directly act as neurotransmitters

e.g. glycine aspartate, glutamate.

• Protein metabolism is more appropriately learnt as metabolism of

amino acids.
Overview of body’s amino acid pool – Sources & utilization
 Metabolism of amino acids —General aspects
• The amino acids undergo certain common reactions
like transamination followed by deamination for
the liberation of ammonia.
• The amino group of the amino acids is utilized for
the formation of urea which is an excretory end
product of protein metabolism.
• The carbon skeleton of the amino acids is first
converted to keto acids (by transamination) which
meet one or more of the following fates.

1. Utilized to generate energy.

2. Used for the synthesis of glucose.

3. Diverted for the formation of fat or ketone bodies.

4. Involved in the production of non-essential amino

acids.
 Transamination
• The transfer of an amino (NH2) group from an amino acid to a keto acid
is known as transamination.
• This process involves the interconversion of a pair of amino acids and a pair
of keto acids, catalysed by a group of enzymes called transaminases
(recently, aminotransferases).
 Salient features of transamination
1. All transaminases require pyridoxal phosphate (PLP), a coenzyme
derived from vitamin B6.

2. Specific transaminases exist for each pair of amino and keto acids.
However, only two— namely, aspartate transaminase and alanine
transaminase—make a significant contribution for transamination.

3. There is no free NH3 liberated, only the transfer of amino group occurs.

4. Transamination is reversible.

5. Transamination is very important for the redistribution of amino groups

and production of non-essential amino acids, as per the requirement of
the cell. It involves both catabolism (degradation) and anabolism
(synthesis) of amino acids.
 Salient features of transamination
6. Transamination diverts the excess amino acids towards energy
generation.

7. The amino acids undergo transamination to finally concentrate nitrogen

in glutamate. Glutamate is the only amino acid that undergoes oxidative
deamination to a significant extent to liberate free NH3 for urea synthesis.

8. All amino acids except lysine, threonine, proline and hydroxyproline

participate in transamination.

9. Transamination is not restricted to α-amino groups only. For instance,

α-amino group of ornithine is transaminated.

10. Serum transaminases are important for diagnostic and prognostic

purposes.
Mechanism of transamination

Transamination occurs in two stages

1. Transfer of the amino group to the

coenzyme pyridoxal phosphate
(bound to the coenzyme) to form
pyridoxamine phosphate.

2. The amino group of pyridoxamine

phosphate is then transferred to a
keto acid to produce a new amino
acid and the enzyme with PLP is
regenerated.
 Mechanism of transamination
• All the transaminases require pyridoxal
phosphate (PLP), a derivative of vitamin B6.
• The aldehyde group of PLP is linked with α-amino
group of lysine residue, at the active site of the
enzyme forming a Schiff base (imine linkage).
• When an amino acid (substrate) comes in contact
with the enzyme, it displaces lysine and a new
Schiff base linkage is formed.
• The amino acid-PLP-Schiff base tightly binds
with the enzyme by noncovalent forces.
• Snell and Braustein proposed a Ping Pong Bi Bi
mechanism involving a series of intermediates
(aldimines and ketimines) in transamination
reaction.
 Deamination

• The removal of amino group from the amino acids as NH3 is

deamination.

• Transamination involves only the shuffling of amino groups

among the amino acids. On the other hand, deamination results

in the liberation of ammonia for urea synthesis. Simultaneously,

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

• Deamination may be either oxidative or non-oxidative.

 I. Oxidative deamination

 Oxidative deamination is the liberation of free ammonia from

the amino group of amino acids coupled with oxidation.

 This takes place mostly in liver and kidney.

 The purpose of oxidative deamination is to provide NH3 for urea

synthesis and α-keto acids for a variety of reactions, including

energy generation.
 I. Oxidative deamination
Role of glutamate dehydrogenase:
•In the process of transamination, the amino groups of most amino acids are transferred
to α-ketoglutarate to produce glutamate. Thus, glutamate serves as a ‘collection centre’
for amino groups in the biological system.
•Glutamate rapidly undergoes oxidative deamination, catalyzed by glutamate
dehydrogenase (GDH) to liberate ammonia. This enzyme is unique in that it can utilize
either NAD+ or NADP + as a coenzyme.
•Conversion of glutamate to α-ketoglutarate occurs through the formation of an
intermediate, α-iminoglutarate.
 I. Oxidative deamination
Oxidative deamination by amino acid oxidases:
•L-Amino acid oxidase and D-amino acid oxidase are flavoproteins, possessing FMN and FAD,
respectively. They act on the corresponding amino acids (L or D) to produce α-keto acids and
NH3. In this reaction, oxygen is reduced to H2O2, which is later decomposed by catalase.

•The activity of L-amino acid oxidase is much low while that of D-amino acid oxidase is high in
tissues (mostly liver and kidney).
•L-Amino acid oxidase does not act on glycine and dicarboxylic acids. This enzyme, due to its
very low activity, does not appear to play any significant role in the amino acid metabolism.
 II. Non-oxidative deamination
• Some of the amino acids can be deaminated to liberate NH 3 without undergoing
oxidation

(a) Amino acid dehydrases: Serine, threonine and homoserine are the hydroxy amino
acids. They undergo non-oxidative deamination catalysed by PLP-dependent
dehydrases (dehydratases).

(b) Amino acid desulfhydrases: The sulfur amino acids, namely cysteine and
homocysteine, undergo deamination coupled with desulfhydration to give keto acids.

(c) Deamination of histidine: The enzyme histidase acts on histidine to liberate NH 3 by a

non-oxidative deamination process.
 Urea cycle
• Urea is the end product of protein metabolism (amino acid metabolism).
 The nitrogen of amino acids, converted to ammonia, is toxic to the body.

 It is converted to urea and detoxified.

 As such, urea accounts for 80-90% of the nitrogen containing substances

excreted in urine.
• Urea is synthesized in liver and transported to kidneys for excretion in
urine.
• Urea cycle is the first metabolic cycle that was elucidated by Hans Krebs
and Kurt Henseleit (1932), hence it is known as Krebs-Henseleit cycle.
 Urea cycle

• Urea has two amino (NH2)

groups, one derived from NH3

and the other from aspartate.
Carbon atom is supplied by
CO2.

• Urea synthesis is a five-step

cyclic process, with five distinct
enzymes.
 The first two enzymes are
present in mitochondria while
the rest are localized in cytosol.
 Urea cycle
1. Synthesis of carbamoyl
phosphate
2. Formation of citrulline
3. Synthesis of arginosuccinate
4. Cleavage of arginosuccinate
5. Formation of urea
 1. Synthesis of carbamoyl phosphate

• Carbamoyl phosphate synthase I

(CPS I) of mitochondria catalyses
the condensation of NH4+ ions with

CO2 to form carbamoyl phosphate.

• This step consumes two ATP and is

irreversible, and rate-limiting. CPS
I requires N-acetylglutamate for its
activity.
• Another enzyme, carbamoyl
phosphate synthase II (CPS II)—
involved in pyrimidine synthesis—is
present in cytosol. It accepts amino
group from glutamine and does not
require N-acetylglutamate for its
activity.
 2. Formation of citrulline
• Citrulline is synthesized from
carbamoyl phosphate and ornithine by
ornithine transcarbamoylase.
• Ornithine is regenerated and used in
urea cycle. Therefore, its role is
comparable to that of oxaloacetate in
citric acid cycle.
• Ornithine and citrulline are basic
amino acids. (They are never found in
protein structure due to lack of
codons).
• Citrulline produced in this reaction is
transported to cytosol by a transporter
system.
 3. Synthesis of arginosuccinate

• Arginosuccinate synthase
condenses citrulline with
aspartate to produce
arginosuccinate.
• The second amino group of urea
is incorporated in this reaction.
• This step requires ATP which is
cleaved to AMP and
pyrophosphate (PPi). The latter is
immediately broken down to
inorganic phosphate (Pi).
 4. Cleavage of arginosuccinate

• Arginosuccinase cleaves
arginosuccinate to give
arginine and fumarate.
• Arginine is the immediate
precursor for urea.
• Fumarate liberated here
provides a connecting link
with TCA cycle,
gluconeogenesis etc.
 5. Formation of urea
• Arginase is the fifth and final enzyme that
cleaves arginine to yield urea and ornithine.
• Ornithine, so regenerated, enters
mitochondria for its reuse in the urea cycle.
• Arginase is activated by Co2+ and Mn2+.

• Ornithine and lysine compete with arginine

(competitive inhibition).
• Arginase is mostly found in the liver,
while the rest of the enzymes (four) of urea
cycle are also present in other tissues.
• For this reason, arginine synthesis may
occur to varying degrees in many tissues.
But only the liver can ultimately produce
urea.
 Urea cycle – Overall reaction and energetics

• The urea cycle is irreversible and consumes 4 ATP.

• Two ATP are utilized for the synthesis of carbamoyl phosphate.

• One ATP is converted to AMP and PPi to produce

arginosuccinate which equals to 2 ATP.
• Hence 4 ATP are actually consumed.

NH4+ + CO2 + Aspartate + 3ATP Urea + Fumarate + 2 ADP + 2 Pi + AMP + PPi

 Metabolic disorders of urea cycle

• Metabolic defects associated with each of the five enzymes of urea cycle have been
reported.
• All the disorders invariably lead to a build-up in blood ammonia (hyperammonemia),
leading to toxicity.
• Other metabolites of urea cycle also accumulate which, however, depends on the
specific enzyme defect.
• The clinical symptoms associated with defect in urea cycle enzymes include vomiting,
lethargy, irritability, ataxia and mental retardation.
Metabolic defects in urea cycle
Defect Enzyme involved
Hyperammonemia type I Carbamoyl phosphate synthase I
Hyperammonemia type II Ornithine transcarbamoylase
Citrullinemia Arginosuccinate synthase
Arginosuccinic aciduria Arginosuccinase
Hyperargininemia Arginase