Amino acid metabolism

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 which meet one or more of the
following fates.

• Utilized to generate energy.

• Used for the synthesis of glucose.
• Diverted for the formation of fat or ketone bodies.
• Involved in the production of nonessential amino acids.

FIG. 15.2 An overview of amino acid metabolism.

The details of general and specific metabolic reactions of amino acids are
Transamination described in the following pages.

The transfer of an amino group from an amino acid to a keto acid is known as
Transamination
transamination. This process involves
The transfer the(−NH
of an amino interconversion of a pair of amino acids and a pair
2) group from an amino acid to a keto acid is
of keto acids, catalysed by as
known a group of enzymes
transamination. called
This process transaminases
involves (aminotransferases).
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
Salient features of transamination
• All transaminases require pyridoxal
1. All transaminases phosphate
require pyridoxal (PLP),
phosphate (PLP), a coenzyme
a coenzyme derived derived from
from vitamin B6.
vitamin B6. 2. Specific transaminases exist for each pair of amino and keto acids. However,
• only two—exist
Specific transaminases namely,for
aspartate
eachtransaminase
pair of and alanineand
amino transaminase—make
keto acids. However, only
a significant contribution for transamination.
two— namely, aspartate transaminase and alanine transaminase—make a significant
contribution for transamination.
• There is no free NH3 liberated, only the transfer of amino group occurs.
• Transamination is reversible
 3. There is no free NH3 liberated, only the transfer of amino group occurs.
4. Transamination is reversible (Fig.15.3).

FIG. 15.3 Transamination reaction.

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

• Transamination production
is very important
of nonessentialforaminothe redistribution
acids, of amino
as per the requirement of the cell.groups
It and production
of nonessential involves
amino bothacids,
catabolismas per theandrequirement
(degradation) of ofthe
anabolism (synthesis) aminocell. It involves both
acids.
catabolism (degradation)
6. Transaminationanddivertsanabolism
the excess amino (synthesis) of amino
acids towards energy acids.
generation.
• Transamination diverts
7. The the
amino acids excess
undergo amino
transamination acids towards
to finally concentrateenergy
glutamate. Glutamate is the only amino acid that undergoes oxidative
nitrogen ingeneration.

• The amino acidsdeamination

undergo to atransamination to finally
significant extent to liberate concentrate
free NH for
3 urea synthesis. nitrogen in glutamate.
Glutamate is the
8. Allonly
aminoamino acid
acids except that
lysine, undergoes
threonine, proline andoxidative
hydroxyprolinedeamination to a significant
participate in transamination.
extent to liberate free NH3is for
9. Transamination urea synthesis.
not restricted to α-amino groups only. For instance, δ-
amino group of ornithine is transaminated.
• All amino acids except
10. Serum lysine,are threonine,
transaminases proline
important for diagnostic and hydroxyproline
and prognostic purposes. participate in
transamination. (Refer Chapter 6).
• Transamination is not restricted
Mechanism to α-amino groups only. For instance, δ-amino group
of transamination
of ornithine isTransamination
transaminated. occurs in two stages (Fig.15.4)

• Serum transaminases are important for diagnostic and prognostic purposes.

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.

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
NADP as a coenzyme. Conversion of glutamate to α-ketoglutarate occurs
reactions, including through
energy the generation.
formation of an intermediate, α-iminoglutarate (Fig.15.5).

FIG. 15.5 Oxidation of glutamate by glutamate dehydrogenase (GDH).

Glutamate dehydrogenase catalysed reaction is important as it reversibly links

Non-oxidative deamination
up glutamate metabolism with TCA cycle through α-ketoglutarate. GDH is
involved in both catabolic and anabolic reactions.
Some of the amino Regulation
acids canof be deaminated to liberate NH3 without undergoing oxidation.
GDH activity
Amino acid dehydrases : Serine,
Glutamate threonine
dehydrogenase and
is a zinc homoserine
containing are the
mitochondrial hydroxy
enzyme. It is a amino acids. They
complex enzyme consisting of six identical units with a molecular weight of
undergo non-oxidative deamination catalysed by PLP dependent dehydrases (dehydratases).
56,000 each. GDH is controlled by allosteric regulation. GTP and ATP inhibit—
whereas GDP and ADP activate—glutamate dehydrogenase. Steroid and thyroid
hormones inhibit GDH.
After ingestion of a protein-rich meal, liver glutamate level is elevated. It is
converted to α-ketoglutarate with liberation of NH3. Further, when the cellular
energy levels are low, the degradation of glutamate is increased to provide α-
ketoglutarate which enters TCA cycle to liberate energy.
 II Non-oxidative deamination
SomeSome of of
thetheamino
aminoacids
acidscan
can be
be deaminated
deaminated totoliberate
liberateNH
NHwithout
3 without undergoing
undergoing
3
oxidation
oxidation
(a) (a)
Amino
Amino acid
aciddehydrases
dehydrases:: Serine, threonineand
Serine, threonine andhomoserine
homoserine areare
the the hydroxy
hydroxy
amino
aminoacids.
acids.They
Theyundergo
undergo non-oxidative deamination
non-oxidative deamination catalysed
catalysed by by
PLPdependentdehydrases
PLPdependent dehydrases (dehydratases).
(dehydratases).

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

Deamination of histidine : The enzyme histidase acts on histidine to liberate NH3 by a non-
(c) Deamination of histidine : The enzyme histidase acts on histidine to liberate
oxidative deamination process.
NH by a non-oxidative deamination process.
3

Decarboxylation Metabolism of ammonia

Ammonia is constantly being liberated in the metabolism of amino acids
(mostly) and other nitrogenous compounds. At the physiological pH, ammonia
Some of the α-amino
exists asacids undergo
ammonium ( decarboxylation to form the respective amines. This
) ion.

is carried out by a group Iof enzymes

Formation called decarboxylases which are dependent on PLP. Many
of ammonia
biogenic amines with important functions
The production arefrom
of NH occurs 3 synthesized
the amino acidsby PLP decarboxylation.
(transamination and
deamination), biogenic amines, amino group of purines and pyrimidines and by
the action of intestinal bacteria (urease) on urea.
Serotonin (5-hydroxytryptamine,
II Transport and storage of NH
5HT), produced from tryptophan is important in
3
nerve impulse transmission
Despite (neurotransmitter). It regulates
a regular and constant production sleep,
of NH from various3
behaviour,
tissues, its blood pressure
etc. concentration in the circulation is surprisingly low (normal plasma 10–20
mg/dl). This is mostly because the body has an efficient mechanism for NH 3
transport and its immediate utilization for urea synthesis. The transport of
ammonia between various tissues and the liver mostly occurs in the form of
glutamine or alanine and not as free ammonia. Alanine is important for NH3
transport from muscle to liver by glucose-alanine cycle (Refer Fig.13.13).

Role of glutamine
Glutamine is a storehouse of NH3. It is present at the highest concentration (8
mg/dl in adults) in blood among the amino acids. Glutamine serves as a storage
and transport form of NH3. Its synthesis mostly occurs in liver, brain and muscle.
Ammonia is removed from the brain predominantly as glutamine. Glutamine is
Urea cycle freely diffusible in tissues, hence easily transported.

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. The individual reactions, however, were described
in more detail later on by Ratner and Cohen. 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.
 synthesis.
3. Citric acid cycle is an important metabolic pathway for the complete oxidation
of various metabolites to CO2 and H2O. The CO2 liberated in TCA cycle (in
the mitochondria) can be utilized in urea cycle.

Metabolic disorders of urea cycle

Metabolic defects associated with each of the five enzymes of urea cycle have
Metabolic disorders of urea cycle
been reported (Table 15.1). All the disorders invariably lead to a build-up in
blood ammonia (hyperammonemia), leading to toxicity. Other metabolites of
Metabolic defects associated
urea cycle with
also each of thewhich,
accumulate five enzymes
however,of urea cycle
depends on thehave beenenzyme
specific
reported. All the disorders invariably
defect. Thelead to a build-up
clinical symptoms in associated
blood ammonia (hyperammonemia),
with defect in urea cycle enzymes
leading to toxicity. The clinical symptoms
include associated
vomiting, with defectataxia
lethargy, irritability, in urea
andcycle enzymes
mental include
retardation.
vomiting, lethargy, irritability, ataxia and mental retardation.
Table 15.1
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

Blood
Catabolism of Phenylalanine and urea—clinical
tyrosine importance
In healthy people, the normal blood urea concentration is 10-40 mg/dl. Higher
protein intake marginally increases blood urea level, however this is well within
normal range. About 15–30 g of urea (7–15 g nitrogen) is excreted in urine per
day.
Blood urea estimation is widely used as a screening test for the evaluation of
kidney (renal) function. It is estimated in the laboratory either by urease method
or diacetyl monoxime (DAM) procedure. Elevation in blood urea may be
broadly classified into three categories.
 Phenylalanine and tyrosine are structurally related aromatic amino acids. Phenylalanine
is an essential amino acid while tyrosine is nonessential. Besides its incorporation into proteins,
the only function of phenylalanine is its conversion to tyrosine. The predominant metabolism
of phenylalanine occurs through tyrosine. Tyrosine is incorporated into proteins and is involved
in the synthesis of a variety of biologically important compounds epinephrine, norepinephrine,
dopamine (catecholamines), thyroid hormones and the pigment melanin. During the course of
degradation, phenylalanine and and
urinary oxalate tyrosine are converted
the formation to stones.
of oxalate metabolites which can serve as
precursors for the Blood
synthesis of glucose and fat. Hence, these aminorenal
urea estimation is commonly used to assess acidsfunction.
are both glucogenic
and ketogenic. Elevation of blood urea level (normal 10–40 mg/dl) is associated
with several disorders which may be prerenal (diabetic coma),
renal (acute glomerulonephritis) and post-renal (tumors or stones
in the urinary tract).
Estimation of serum creatinine (normal < 1 mg/dl) is considered
to be a more reliable indicator for the evaluation of kidney
function.

Conversion of phenylalanine to tyrosine

Under normal circumstances, the degradation of phenylalanine mostly occurs
through tyrosine. Phenylalanine is hydroxylated at para-position by
phenylalanineFIG.hydroxylase
15.17 Overviewto of phenylalanine and tyrosine
produce tyrosine metabolism (CNS–
(phydroxy phenylalanine). This
Central nervous system; T3–Triiodothyronine).
is an irreversible reaction
Conversion of phenylalanine to tyrosine and requires the participation of a specific coenzyme
biopterin (containing pteridine ring) which is structurally related to folate. The
active
Under formcircumstances,
normal of biopterintheisdegradation
tetrahydrobiopterin (H4-biopterin).
of phenylalanine In the
mostly occurs through
phenylalanine ishydroxylase
tyrosine. Phenylalanine hydroxylatedreaction, tetrahydrobiopterin
at para-position by phenylalanineis oxidized to
hydroxylase to
dihydrobiopterin (H -biopterin).
produce tyrosine. This is an irreversible
2 Tetrahydrobiopterin is then regenerated by
reaction and requires the participation of a specific an
biopterin which is Biomedical/clinical
coenzyme NADPH-dependent dihydrobiopterin
structurally relatedreductase concepts
to folate.(Fig.15.18).
About 300–400 g of protein per day is constantly degraded and
synthesized in the human body.
The amino acids are mainly utilized for protein biosynthesis,
production of specialized products (creatine, porphyrin, amines,
purines, pyrimidines) and generation of energy.
Glutamate is the collection centre for the amino groups in the
biological system while glutamine is the storehouse of NH3. Free
NH3 can be liberated predominantly from glutamate.
Ammonia accumulation in blood is toxic to brain causing slurring
of speech, blurring of vision, tremors and even death. Mammals
convert
FIG. 15.18NH 3 to urea,ofa tyrosine
Synthesis non-toxic excretory
from product.
phenylalanine ( Metabolic
—Block in
phenylketonuria).
defects in urea cycle enzymes result in hyperammonemia.
Degradation of tyrosine (phenylalanine)
Dietary consumption of a protein rich meal increases the level of
The enzyme phenylalanine
N-acetylglutamate in liverhydroxylase
which enhances is urea
present in the liver. In the
production.
conversion Primary
of phenylalanine to tyrosine,
hyperoxaluria—a thedisorder
metabolic reaction
dueinvolves
to a defectthe
in incorporation
the enzyme glycine transaminase— is characterized by elevated
of one atom of molecular oxygen (O2) into the para position of phenylalanine
The metabolism of phenylalanine and tyrosine is considered together. The
sequence of the reactions in the degradation of these amino acids, depicted in
Fig.15.19, is described hereunder

FIG. 15.19 Tyrosine metabolism—degradative pathway [αKG–α-

Ketoglutarate; Glu-Glutamate; The circled numbers indicate metabolic
Disorders of tyrosine (phenylalanine) metabolism
defects (1) Phenylketonuria; (2) Tyrosinemia type II; (3) Neonatal
tyrosinemia; (4) Alkaptonuria; (5) and (6) Tyrosinosis (tyrosinemia, type I)].
Phenylketonuria

Phenylketonuria (PKU) is the most common metabolic disorder in amino acid

metabolism. The incidence of PKU is 1 in 10,000 births. It is due to the deficiency of the hepatic
enzyme, phenylalanine hydroxylase, caused by an autosomal recessive gene. Phenylketonuria
primarily causes the accumulation of phenylalanine in tissues and blood, and results in its
increased excretion in urine. Due to disturbances in the routine metabolism, phenylalanine is
diverted to alternate pathways, resulting in the excessive production of phenylpyruvate,
phenylacetate, phenyllactate and phenylglutamine. All these metabolites are excreted in urine
in high concentration in PKU. Phenylacetate gives the urine a mousey odour. The name
phenylketonuria is coined due to the fact that the metabolite phenylpyruvate is a keto acid
excreted in urine in high amounts.

Tyrosinemia type II

This disorder also known as Richner-Hanhart syndrome, is due to a defect in the

enzyme tyrosine transaminase. The result is a blockade in the routine degradative pathway of
tyrosine. Accumulation and excretion of tyrosine and its metabolites namely p-
hydroxyphenylpyruvate, p-hydroxyphenyllactate, p-hydroxyphenylacetate, N-acetyltyrosine
and tyramine are observed. Tyrosinemia type II is characterized by skin (dermatitis) and eye
lesions and, rarely, mental retardation. A disturbed self-coordination is seen in these patients.

Neonatal tyrosinemia

The absence of the enzyme p-hydroxyphenylpyruvate dioxygenase causes neonatal

tyrosinemia. This is mostly a temporary condition and usually responds to ascorbic acid. It is
explained that the substrate inhibition of the enzyme is overcome by the presence of ascorbic
acid.

Alkaptonuria (Black urine disease)

Alkaptonuria has great historical importance. It was first described by Lusitanus in 1649
and characterized in 1859. The defective enzyme in alkaptonuria is homogentisate oxidase in
tyrosine metabolism. Homogentisate accumulates in tissues and blood, and is excreted into
urine. Homogentisate, on standing, gets oxidized to the corresponding quinones, which
polymerize to give black or brown colour. For this reason, the urine of alkaptonuric patients
resembles coke in colour.

Tyrosinosis or tyrosinemia type I

This is due to the deficiency of the enzymes fumarylacetoacetate hydroxylase and/or

maleylacetoacetate isomerase. Tyrosinosis is a rare but serious disorder. It causes liver failure,
rickets, renal tubular dysfunction and polyneuropathy. Tyrosine, its metabolites and many other
amino acids are excreted in urine. In acute tyrosinosis, the infant exhibits diarrhea, vomiting,
and ‘cabbage-like’ odor. Death may even occur due to liver failure within one year. For the
treatment, diets low in tyrosine, phenylalanine and methionine are recommended.

Albinism

Albinism (Greek: albino—white) is an inborn error, due to the lack of synthesis of the pigment
melanin. The melanin synthesis can be influenced by a variety of factors. Many possible causes
for albinism have been identified

• Deficiency or lack of the enzyme tyrosinase.

• Decrease in melanosomes of melanocytes.
• Impairment in melanin polymerization.
• Lack of protein matrix in melanosomes.
• Limitation of substrate (tyrosine) availability.
• Presence of inhibitors of tyrosinase.

The most common cause of albinism is a defect in tyrosinase, the enzyme most responsible
for the synthesis of melanin.

Biosynthesis of catecholamines

The name catechol refers to the dihydroxylated phenyl ring. The amine derivatives of
catechol are called catecholamines. Tyrosine is the precursor for the synthesis of
catecholamines, namely dopamine, norepinephrine (noradrenaline) and epinephrine
(adrenaline).

FIG. 15.22 Metabolism of tyrosine-synthesis of catecholamines

(dopamine, norepinephrine, epinephrine; PLP–pyridoxal phosphate).
Functions of catecholamines
Tyrosine is hydroxylated to 3,4-dihydroxyphenylalanine (DOPA) by tyrosine
hydroxylase.
Norepinephrine and Thisepinephrine
enzyme catalyses the carbohydrate
regulate rate limiting reaction
and lipidand requires
metabolisms. They
tetrahydrobiopterin as coenzyme (like phenylalanine hydroxylase). In contrast
stimulate the degradation of triacylglycerol and glycogen. They cause an increase in to the blood
pressure. Dopamine and norepinephrine serve as neurotransmitters in the brain and
autonomous nervous system.

Serotonin

Serotonin or 5-hydroxytryptamine (5HT) is a neurotransmitter, synthesized from

tryptophan. Normally, about 1% of the tryptophan is converted to serotonin.

Functions of serotonin

Serotonin is a neurotransmitter and performs a variety of functions.

 • Serotonin is a powerful vasoconstrictor and results in smooth muscle contraction in
bronchioles and arterioles.
• It is closely involved in the regulation of cerebral activity (excitation).
• Serotonin controls the behavioural patterns, sleep, blood pressure and body
temperature.
• Serotonin evokes the release of peptide hormones from gastrointestinal tract.
• It is also necessary for the motility of GIT (peristalsis).

Melatonin

Melatonin is a hormone, mostly synthesized by the pineal gland. Serotonin produced

from tryptophanis acted upon by serotonin N-acetylase to give N-acetylserotonin. The latter
undergoes methylation, S-adenosylmethionine being the methyl group donor to produce
melatonin or N- acetyl 5-methoxyserotonin

Functions of melatonin

• Melatonin is involved in circadian rhythms or diurnal variations of the body. It plays a

significant role in sleep and wake process.
• Melatonin inhibits the production of melanocyte stimulating hormone (MSH) and
adrenocorticotropic hormone (ACTH).
• It has some inhibitory effect on ovarian functions.
• Melatonin also performs a neurotransmitter function.

Catabolism of Heme

When haemoglobin is destroyed in the body, globin is degraded to its constituent amino
acids, which are reused and the iron of heme enters the iron pool The catabolism of heme is
carried out in the microsomal fraction of cell by a complex enzyme system called heme
oxygenase. It is located in close proximity to the microsomal electron transport system. The
enzyme heme oxygenase adds a hydroxyl group to the methenyl bridge between the two pyrrole
rings with a concomitant oxidation of ferrous iron to Fe+3 in the presence of NADPH and O2.
A second oxidation results in cleavage of the porphyrin ring. Ferric ion and CO2 are released
yielding green pigment biliverdin which is reduced by the enzyme biliverdin reductase, using
NADPH as reductant, forming red orange bilirubin.
Jaundice: The normal serum total bilirubin concentration is in the range of 0.2 to 1.0 mg/dl.
Of this, about 0.2-0.6 mg/dl is unconjugated while 0.2 to 0.4 mg/dl is conjugated bilirubin.
Jaundice (French : Jaune-yellow) is a clinical condition characterized by yellow colour of the
white of the eyes (sclerae) and skin. It is caused by the deposition of bilirubin due to its elevated
levels in the serum. The term hyperbilirubinemia is often used to represent the increased
concentration of serum bilirubin.