1. Dr. Saidunnisa, MD Professor of Biochemistry
Aromatic amino acid (phenylalanine and tyrosine )metabolism
and its inborn errors.

2. Learning Objectives
At the end of the session the student shall be able to:
Explain the major and minor pathways of phenylalanine and tyrosine metabolism.
Interpret the biochemical aspects of disorders like PKU and Tyrosinemia and Alkaptunuria associated with them.
Explain the melanin pathway with albinism
Explain the neurotransmitters and thyroid hormone metabolic pathway
Interpret parkinsonism, goiter ,hyper and hypothyroidism.

3. Overview Phenylalanine and tyrosine metabolism: Major pathway
Minor pathway
Disorders:
Phenyl Ketonuria
Tyrosinemia
Alkaptonuria
Albinism
Parkinsonism,
Hyper and hypothyroidism.

4. Phenylalanine and tyrosine Metabolism
Phenylalanine is an essential
Tyrosine is an non-essential amino acid.
These are both glucogenic and ketogenic amino acids.
The Predominant metabolism of phenylalanine occurs through tyrosine.
These are involved in a variety of biologically important compounds:
Epinephrine, nor-epinephrine, dopamine, (catecholamine)
Melanin
Thyroid hormones

5. Overview- Major and minor pathway

6. Major route of phenylalanine metabolism

7. Conversion of phenylalanine to tyrosine
Phenylalanine is hydroxylated at Para- position by phenylalanine hydroxylase to produce tyrosine.
This is an irreversible reaction.
It requires a specific coenzyme biopterin.

8. Mechanism of the reaction
Phenylalanine hydroxylase is present in liver.
involves incorporation of one atom of molecular oxygen into para position of phenylalanine while the other atom of oxygen is reduced to water (Monoxygenase).

9. Mechanism of the reaction
The active form of biopterin is H4-biopterin.
In this reaction H4- biopterin is oxidized to H2- biopeterin.
H4-biopterin is then regenerated by an NADPH-dependent dihydrobiopterin reductase.

10. Inborn Errors of Metabolism
An inherited enzyme deficiency caused by mutant genes, leading to the disruption of normal bodily metabolism
Accumulation of a toxic substrate (compound acted upon by an enzyme in a chemical reaction)
Impaired formation of a product normally produced by the deficient enzyme.

11. What is a metabolic disease?
Garrod’s hypothesis
product deficiency
substrate excess
toxic metabolite C A B D

12. Inborn errors of metabolism
More than 50 of these disorders have been described.
Collectively they constitute a very significant portion of pediatric genetic diseases.
Phenylketonuria is the most common disease.

13. Phenylketonuria
In the United States, PKU is detected in 1/10,000 (Caucasian population) to 1/50,000 (African American population) newborns.
Phenylketonuria (PKU), caused by a deficiency of phenylalanine hydroxylase.
Biochemically, it is characterized by accumulation of phenylalanine .
Hyperphenylalaninemia may also be caused by deficiencies in any of the several enzymes required to synthesize BH4, or in dihydropteridine (BH2) reductase,

14. Due to the deficiency phenylalanine is converted to phenylpyruvate and this is further converted to Phenyl lactate, and to phenyl acetate.
Minor pathway
Due to defect in phenylalanine hydroxylase, the conversion of phenylalanine to tyrosine is blocked resulting in high levels of phenylalanine in the body.

15. Characteristics of classic PKU:
Phenylalanine is present in elevated concentrations in tissues, plasma, and urine.
Phenyllactate, phenylacetate, and phenylpyruvate, which are not normally produced in significant amounts in the presence of functional phenylalanine hydroxylase, are also elevated in PKU.
[Note: The disease acquired its name from the presence of a phenylketone (now known to be phenylpyruvate) in the urine.

16. Signs and symptoms: Mental retardation, failure to walk or talk,
seizures,
hyperactivity,
tremor,
microcephaly, and
failure to grow
These metabolites give urine a characteristic musty (“mousey”) odor.
[Note: These clinical manifestations are now rarely seen as a result of neonatal screening programs.]

17. Biochemical basis for CNS symptoms
Accumulation of phenylalanine in brain impairs the transport and metabolism of other aromatic amino acids (tryptophan, tyrosine).
The synthesis of serotonin an excitatory neurotransmitter from tryptophan is insufficient.

18. Learning check
Can PKU cause hypo pigmentation give the biochemical reasoning?

19. Albinism

20. Diagnosis
Ferric chloride test: (Urine)qualitative test gives green colour due to presence of phenylpyruvic acid.
Guthrie test: which is a bacterial (bacillus subtilis) bioassay:
for phenylalanine.
Normal levels:
1-2mg/dl.

21. Treatment
Maintenance of plasma phenylalanine concentration within normal limits is a challenging task in treatment of PKU.
This is done by strictly limiting dietary intake of phenylalanine and supplementing intake of tyrosine.
Most experts believe that the diet should be adhered to throughout life but this can be very difficult, especially during adolescence and early adulthood.

22. Some foods PKU patients should restrict
This diet essentially excludes milk and dairy products, meat, fish, chicken, eggs, beans and nuts which all contain very high levels of phenylalanine.
Fruits, vegetables, breads and pastas also contain some phenylalanine and cannot be eaten freely.
Soft drinks and foods containing the sweetener aspartame must also be avoided.

23. Tyrosinemia This disorder is also known as Richner-Hanhart syndrome.
Deficiency of tyrosine transaminase.
Accumulation of tyrosine and its metabolites Phenyllactate, phenylacetate, and phenylpyruvate.
Characterized by:
dermatitis,
eye lesions,
mental retardation.

24. Alkaptonuria
Alkaptonuria is a rare metabolic disease involving a deficiency in Homogentisic acid oxidase, resulting in the accumulation of Homogentisic acid.

25. Conversion of Homogentisate to Alkapton
Poly phenol oxidase
Benzoquinone acetate
Polymerization
Alkapton
Binds to tissues: of connective tissue, bones and various organs (nose, ear etc) resulting in a condition known as Ochronosis.

26. Signs of Alkaptonuria

27. Ochronosis
Dark staining of the diapers sometimes can indicate the disease in infants, but usually no symptoms are present until later in life.
Change in colour of the urine on standing to brown or dark.

28. Alkaptonuria symptoms
The illness has three characteristic symptoms:
homogentisic aciduria the patient's urine contains elevated levels of Homogentisic acid,
Which is oxidized to a dark pigment on standing, large joint arthritis, and
Black ochronotic pigmentation of cartilage and collagenous tissue.

29. Diagnosis Change in colour of the urine on standing to brown or dark.
Urine gives positive with ammonical silver nitrate.
This is due to strong reducing activity of homogentisate.

30. Treatment Diets low in phenylalanine and tyrosine:
help in reducing the levels of Homogentisic acid, and decrease the amount of pigment deposited in body tissues.
Although Alkaptonuria is not life- threatening, the associated arthritis may be severely crippling.

31. Melanin synthesis
Melanin is a pigment that occurs in several tissues, particularly the eye, hair, and skin.
It is synthesized from tyrosine in the epidermis by pigment-forming cells called melanocytes.
Its function is to protect underlying cells from the harmful effects of sunlight.

32. Albinism
Autosomal recessive
A defect in melanin production results in albinism, the most common form being due to defect in tyrosinase a copper-containing enzyme.

33. Sings and symptoms
Absence of melanin pigment (hypo pigmentation) from the skin, hair, and eyes.
Affected individuals have vision defects and photophobia (sunlight hurts their eyes). They are at increased risk for skin cancer.
Oculocutaneous albinism having whit eyebrows and lashes.

34. Tyrosine-Derived Neurotransmitters
Tyrosine is converted to the catecholamine's. These are dopamine, nor epinephrine, and epinephrine
These are neurotransmitters .
The first step is conversion to DOPA, by tyrosine hydroxylase, which requires BH4 as cofactor.
DOPA is converted to dopamine.

35. Tyrosine-Derived Neurotransmitters
Dopamine is converted to Norepinephrine and to epinephrine..
Within the substantia nigra and some other regions of the brain, synthesis proceeds only to dopamine.
Within the adrenal medulla dopamine is converted to nor epinephrine and epinephrine.

36. Parkinson’s disease Is commonly seen in elderly people.
Due to degeneration of certain parts of the brain mainly substantia niagra there is decreased production of dopamine.

37. Signs and symptoms Four cardinal features: Remember acronym “TRAP”
Tremor
Rigidity
Akinesia
Postural changes

38. Synthesis of thyroid hormones
Thyroid hormones- thyroxine (T4 tetraiodothronine) and T3 (Triiodothyronine) are synthesized from tyrosine.
Iodination of tyrosine ring occurs to produce mono, di, tri, tetra iodotyrosine in thyroglobulin molecule.
Breakdown of thyroglobulin molecule releases the free hormones.

39. Branched chain amino acids Metabolism
They serve an important role as alternate source of fuel for the brain during starvation.
All the 3 AA are essential.
Valine -Glycogenic
Leucine-Ketogenic
Iso leucine- Glycogenic and Ketogenic

40. Overview of the pathway

41. Pathway The first 3 common Metabolic reactions are:
Transamination: removal of amino groups of all 3AA by PLP requiring transaminase and forming respective ketoacids
Oxidative decarboxylation: removal of carboxyl group from keto acids by the enzyme alpha keto acid dehydrogenase (similar to PDH) are converted to respective acyl CoA esters.
Dehydrogenation: oxidation of acyl CoA esters to succinyl CoA and acetyl CoA (similar to fatty acid oxidation).

42. Maple syrup urine disease (MSUD)
Enzyme defect : is alpha keto acid dehydrogenase (Oxidative decarboxylation step)
Blockade in conversion of alpha ketoacids to Acyl CoA esters.
Plasma and urine has high level of keto acids.
Urine of affected individuals smells like maple syrup or burnt sugar.

43. Symptoms Autosomal recessive disorder. 1: 1,85,000 births Acidosis
Lethargy
Convulsions
Mental retardation
Coma
Death
Treatment: diet low in BCAA

44. Learning check Laboratory diagnosis :
Can Rothera's test be positive for MSUD? Give your explanation.

45. Learning check
What might cause an increase in plasma branched-chain amino acids after 5days of starvation?

46. Answer
BCAA increase from muscle and liver protein breakdown.