Urea cycle

1. Since ammonia is toxic to CNS even in traces liver rapidly removes ammonia from circulation
and converts it to a non-toxic water soluble urea. Hence site of urea synthesis is liver.

2. The reactions leading to formation of urea from ammonia are proposed by Krebs and
Henseleit. Hence, urea cycle is also called as Krebs-Henseleit cycle.

3. Formation of urea from ammonia in urea cycle occurs in five reactions. However the first
reaction is not a part of the cycle but for the continuation of the cycle which consist of
remaining four reactions product of the first reaction is essential..

4. Synthesis of urea from ammonia is a energy dependent process.

5. Enzymes of urea cycle are present in mitochondria and cytosol.

6. First two reactions of urea formation occurs in mitochondria and remaining reactions occur in
cytosol.

Reaction sequence of urea formation: For the synthesis of urea only one ammonia molecule is
used as such. Aspartate serves as donor of another molecule of ammonia. HCO3– serves as
source of CO2 required for urea formation.

1. Formation of carbamoyl phosphate: First reaction leading to urea formation is condensation

of ammonia and HCO3– at the expense of two high-energy bonds to form carbamoyl phosphate.
The reaction is catalyzed by mitochondrial carbamoyl phosphate synthetase-I.

Reactions of urea cycle

2. Now the first reaction of urea cycle is catalyzed by ornithine transcarbamoylase. It condenses
carbamoyl phosphate and ornithine to form citrulline. This enzyme is present in mitochondria.

3. Arginino succinate synthetase present in cytosol catalyzes second reaction of urea cycle. It
condenses citrulline and aspartate at the expense of two high energy bonds to form
argininosuccinate.

4. In the third reaction of urea cycle argininosuccinate is cleaved by argininosuccinase to

arginine and fumarate.
5. Regeneration of ornithine and formation of urea from arginine is the final reaction of urea
cycle. This reaction is catalyzed by arginase.

The ornithine so formed enters mitochondria through specific transporter present in inner
mitochondrial membrane to start reactions of urea cycle once again.

Overall equation for urea formation

NH3 + HCO–3 + Aspartate + 4ATP → Urea + Fumarate + 4ADP + 4Pi

Fate of urea: Urea has no physiological function. Hence it is transported to kidneys where it is
excreted in urine. It is major end product of protein catabolism in humans. About 10-25 gm of
urea is excreted in urine per day which makes up to 80-90% of total nitrogen excreted per day.
However, blood also contains some urea.

Blood urea Normal blood urea level is 16-36 mg/100 ml.

Medical Importance: Urea formation is impaired in several inherited diseases. They are due to
deficiency of enzymes of urea cycle. Since the urea cycle converts ammonia to urea these
disorders of urea cycle cause ammonia intoxication. Some common clinical symptoms seen in
these diseases are vomiting, irritability, lethargy, seizures, mental retardation, coma and early
death. They are

1. Hyper- ammonemia Type I : It is due to deficiency of enzyme carbamoyl phosphate

synthetase-I. Mental retardation is the main symptom of this condition.

2. Hyper- ammonemia Type II: It is most common among others. It is due to deficiency of
enzyme ornithine trans carbamoylase. So, in this condition carbamoyl phosphate accumulates
and diverted to pyrimidine formation. This results in excretion of oroticacid and uracil in urine.
Glutamate also accumulates in this condition.

3. Citrullinemia: This condition is due to the absence of enzyme argininosuccinate synthetase.

Hence citrulline accumulates in blood and excreted in urine.

4.Argininosuccinicaciduria: Argininosuccinase is absent in this condition. So,

argininosuccinate accumulates in blood and excreted in urine.

5. Hyper- argininemia: This condition is due to low arginase activity. Hence, arginine
accumulates and excreted in urine. However some urea may be excreted in urine due to kidney
arginase.

6. N-acetyl glutamate synthetase deficiency: It is a rare disorder. N- acetyl glutamate

synthetase is involved in formation of N- acetyl glutamate from acetyl- CoA and glutamate.
Hyper ammonemia and aminoacid uria occurs in this condition.

Urea production may be decreased in liver diseases

Treatment

Treatment of urea cycle disorders involves removal of excess ammonia from blood and reduction
of dietary nitrogen load. Peritonial dialysis is employed to clear ammonia from blood.
Administration of compounds which can increase nitrogen excretion is another line of treatment.
Benzoic acid and phenyl acetate are two such compounds used in the treatment.
Metabolism of phenyl alanine and Tyrosine: Phenyl alanine is an essential amino acid. Since
tyrosine is a hydroxylated phenyl alanine it is non- essential amino acid. In plants and bacteria,
phenyl alanine and tyrosine are synthesized from erythrose- 4- phosphate and
phosphoenolpyruvate.

Degradation of phenyl alanine and tyrosine: Phenyl alanine and tyrosine are degraded to
fumarate and aceto- acetate. Since degradation of phenylalanine involves first its conversion to
tyrosine, a single pathway is responsible for the degradation of both phenylalanine and tyrosine.

Reaction sequence

1. Conversion of phenylalanine to tyrosine or tyrosine synthesis: First reaction of

phenylalanine catabolism is its hydroxylation to tyrosine, which requires a cofactor which is not
encountered earlier. A tetra hydrobiopterin (THB) requiring phenylalanine hydroxylase catalyzes
this hydroxylation. The enzyme is present in liver and it is a monooxygenase.

2. Now catabolism of tyrosine begins with transamination. p- hydroxy phenyl pyruvic acid is
produced from tyrosine by the action of tyrosine transaminase in this reaction.
3. p-hydroxy phenyl pyruvate hydroxylase a copper containing dioxygenase converts phydroxy
phenyl pyruvic acid to homogentisic acid in a complex reaction involving hydroxylation of
benzene ring, decarboxylation and shifting of side chain.

4. In this reaction, benzene ring of homogentisic acid is cleaved by another dioxygenase called as
homogentisic acid oxidase to form maleyl aceto acetate.

5. A glutathione dependent maleyl aceto acetate cis- trans isomerase isomerizes maleyl aceto
acetate to fumaryl aceto acetate.

6. Finally fumarate and aceto acetate are formed from fumaryl aceto acetate by the action of an
hydrolase.

Thus, four atoms of phenylalanine are released as fumarate, one carbon is released as CO 2 and
remaining four atoms are released as aceto acetate. Fumarate may undergo further catabolism in
TCA cycle.

Biological importance

1. Tyrosine is required for the synthesis of adrenal hormones like epinephrine, norepinephrine
and dopamine.
2. Tyrosine is needed for the formation of melanin.
3. Thyroid hormones T3 and T4 are formed from tyrosine.
4. Tyrosine is a precursor of glucose and fatty acids or ketone bodies.
5. For the formation of proteins tyrosine and phenylalanine are required.
6. In the intestine tyrosine is decarboxylated by microorgnisms to tyramine.
7. Phosphorylation of tyrosine residues of proteins by kinases affects cell growth.