AMINO ACID

METABOLISM
 Proteins are the most abundant organic
compounds & constitute a major part of the
body dry weight (10-12kg in adults).
 Perform a wide variety of structural & dynamic
(enzymes, hormones, clotting factors,
receptors) functions.
 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 & half are
supplied through diet.
 The proteins on degradation release
individual amino acids.
 Each amino acid undergoes its own
metabolism & performs specific functions.
 Some amino acids serve as precursors for
the synthesis of many biologically important
compounds.
 Certain amino acids may directly act as
neurotransmitters (e.g glycine, aspartate,
glutamate)
 About 100g of free amino acids which represent
the amino acid pool of the body.
 Glutamate & glutamine together constitute
about 50% & essential amino acids about 10% of
the body pool (100g).
 The concentration of intracellular amino acids is
always higher than the extracellular amino
acids.
 Enter the cells against a concentration gradient.
 Turnover of body intake of dietary protein &
the synthesis of non-essential amino acids
contribute to the body amino acid pool.
 Protein turnover:
 The proteins in the body is in a dynamic
state.
 About 300-400g of protein per day is constantly
degraded & synthesized which represents
body protein turnover.
 Control of protein turnover:
 The turnover of the protein is influenced by
many factors.
 A small polypeptide called ubiquitin (m.w.8,500)
tags with the protein & facilitates
degradation.
 Certain proteins with amino acid sequence
proline, glutamine, serine & threonine are
rapidly degraded.
 Dietary protein:
 There is a regular loss of nitrogen from the
body due to degradation of amino acids.
 About 30-50g of protein is lost every day.
 This amount of protein is supplied through diet
to maintain nitrogen balance.
 There is no storage form of amino acids in the
body.
 Excess intake of amino acids is oxidized to
provide energy.
 Proteins function as enzymes, hormones,
immunoproteins, contractile proteins
etc.
 Many important nitrogenous compounds
(porphyrins, purines, pyrimidines, etc) are
produced from the amino acids.
 About 10-15% of body energy requirements are
met from the amino acids.
 The amino acids are converted into
 Transamination
 Oxidative Deamination
 Ammonia Transport
 Urea Cycle
 The transfer of an amino (-NH2) group from an
amino acid to a ketoacid, with the formation
of a new amino acid & a new keto acid.
 Catalysed by a group of enzymes
called transaminases
(aminotransferases)
 Pyridoxalphosphate (PLP)– Co-factor.
 Liver, Kidney, Heart, Brain - adequate amount
of these enzymes.
 All transaminases require PLP.
 No free NH3 liberated, only the transfer of
amino group.
 Transamination is reversible.
 There are multiple transaminase enzymes
which vary in substrate specificity.
 AST & ALT make a significant contribution for
transamination.
 Transamination is important for
redistribution of amino groups & production
of non-essential amino acids.
 It diverts excess amino acids towards the
energy generation.
 Amino acids undergo transamination to
finally concentrate nitrogen in glutamate.
 Glutamate undergoes oxidative
deamination to liberate free NH3 for urea
synthesis.
 All amino acids except, lysine, threonine,
proline & hydroxyproline participate in
transamination.
 It involves both anabolism & catabolism,
since – reversible.
AA1 + α- KG ketoacid + 1
Glutamate

Alanine + α- KG Pyruvate + Glutamate

Aspartate + α- Oxaloacetetae +Glutamate

KG
 Step:1
 Transfer of amino group from AA1 to the
coenzyme PLP to form pyridoxamine
phosphate.
 Amino acid1 is converted to Keto acid2.
 Step:2
 Amino group of pyridoxamine phosphate is
then transferred to a keto acid1 to produce a
new AA 2 & enzyme with PLP is
 Enzymes, present within cell, released in
cellular damage into blood.
 ↑ AST - Myocardial Infarction (MI).
 ↑ AST, ALT – Hepatitis, alcoholic cirrhosis.
 Muscular Dystrophy.
 The amino group of most of the amino acids is
released by a coupled reaction, trans-
deamination.
 Transamination followed by oxidative
deamination.
 Transamination takes place in the
cytoplasm.
 The amino group is transported to liver as
glutamic acid, which is finally oxidatively
deaminated in the mitochondria of
hepatocytes.
 The removal of amino group from the amino
acids as NH3 is deamination.
 Deamination results in the liberation of
ammonia for urea synthesis.
 The carbon skeleton of amino acids is
converted to keto acids.
 Deamination may be either oxidative or
non-oxidative
 Only liver mitochondria contain glutamate
dehydrogenase (GDH) which deaminates
glutamate to α-ketoglutarate & ammonia.
 It needs NAD+ as co-enzyme.
 It is an allosteric enzyme.
 It is activated by ADP & inhibited by GTP.
 Oxidative deamination is the liberation of
free ammonia from the amino group of
amino acids coupled with oxidation.
 Site: Mostly in liver & kidney.
 Oxidative deamination is to provide NH3
for urea synthesis & α-keto acids for a
variety of reactions, including energy
generation.
 Glutamate is a 'collection centre' for amino
groups.
 Glutamate rapidly undergoes oxidative
deamination.
 Catalysed by GDH to liberate ammonia.
 It can utilize either NAD+ or NADP+.
 This conversion occurs through the
formation of an α-iminoglutarate
 COO-
I
COO- COO- CH2
I I I
CH2 CH2 CH2
I I I
CH2 NAD(P)+ CH2 CH2 + NH4 +
I GDH I H2O I
GDH
H -C-NH3+ C=NH C=O
I I I
NAD(P)H+H+ COO-
COO- COO-

L-Glutamate α- Iminoglutarate α- ketoglutarate

 Reversible Reaction
 Both Anabolic & Catabolic.
 Regulation of GDH activity:
 Zinc containing mitochondrial, allosteric
enzyme.
 Consists of 6 identical subunits.
 Molecular weight is 56,000.
 GTP & ATP – allosteric inhibitors.
 GDP & ADP - allosteric activators.
 ↓ Energy - ↑ oxidation of A.A.
 Steroid & thyroid hormones inhibit GDH.
 L-amino acid oxidase & D-Amino acid
oxidase.
 Flavoproteins & Cofactors are FMN &
FAD.
 Act on corresponding amino acids to
produce
α-keto acids & NH3
 Site: Liver, kidney, Peroxisomes.
 Activity of L-Amino acid oxidase is
low.
 L-amino acid
oxidase
L-amino acid α- keto acid + NH3

FMN FMNH2

H2 O2 ½ O2
Catalase

H2O
 L-Amino acid Oxidase acts on all Amino
acids, except glycine & dicarboxylic acids.
 Activity of D-Amino oxidase is high than
that of L-Amino acid oxidase
 D-Amino oxidase degrades D-Amino acids in
bacterial cell wall.
 D-amino acids are found in plants &
microorganisms.
 They are not present in mammalian
proteins.
 D-amino acids are taken in the diet/bacterial
cell wall, absorbed from gut - D-Amino
acid
oxidase converts them to respective α-keto
 The α-ketoacids undergo transamination
to be converted to L-amino acids which
participate in various metabolic pathways.
 Keto acids may be oxidized to generate
energy or serve as precursors for glucose &
fat synthesis.
 H2 O FAD
D-amino acid oxidase
NH4+
FADH2

α-Ketoacid
L-Amino acid
Energ
α- Ketoacid Transaminases y
D-amino acid
L-amino acid Glucose &
Fat
 Direct deamination, without oxidation.
 Amino acid Dehydratases:
 Serine, threonine & homoserine are the
hydroxy amino acids.
 They undergo non-oxidative deamination
catalyzed by PLP-dependent dehydratases
 Serine Dehydratase
Threonine Respective Ketoacid
Homoserine PL
P NH3
Cysteine & homocysteine undergo

deamination coupled with desulfhydration to

give keto acids.
Desulfhydrases
Cysteine Pyruvate
NH3 +H2S

Deamination of histidine:
Histidase
Histidine Uroconate

NH 3
 Urea Cycle
• The cycle is known as Krebs-Henseleit urea
cycle.
• As ornithine is the first member of the
reaction sequences, it is called as Ornithine
cycle.
• The two nitrogen atoms of urea are derived
from two different sources, one from
ammonia and the other directly from
aspartic acid.
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 Steps of Urea Cycle
1. Formation of Carbamoyl Phosphate.
2. Formation of Citrulline.
3. Formation of Argininosuccinate.
4. Formation of Arginine.
5. Formation of Urea.

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 Steps of Urea Cycle
1
2 ATP + HCO3- + NH3 Carbamoyl phosphate + 2 ADP + Pi
Pi
Mitochondrion
. Ornithine 2 Citrulline

Citrulline
Ornithine Urea cycle ATP
3 Aspartate
5 AMP + PPi
Urea Cytosol
H2O Arginino-
Arginine succinate
4

Fumara
te

Malate Oxaloacetate 27
 Disorderers of urea cycle
• Deficiency of any of the urea cycle enzymes
would result in hyperammonemia.
• If block occur in one of the earlier steps, the
condition is more severe, since ammonia itself
accumulates.
• If deficiency occur in later enzymes, this result
in accumulation of other intermediates which
are less toxic and hence symptoms are less.

30
• The accumulation of ammonia in blood
(normally less than 50 mg/dl) and
body fluids results in toxic symptoms.
• Brain is very sensitive to ammonia.
• Child may be put on a low protein diet and
frequent small feeds are given.
• Since Citrulline is present in significant
quantities in milk, breast milk is to be
avoided in Citrullinemia.

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 Urea level in blood and urine
• In clinical practice, blood urea level is taken as an
indicator of renal function.
• The normal urea level in plasma is from 20 to 40
mg/dl.
• Blood urea level is increased where renal function
is inadequate.
• Urinary excretion of urea is 15 to 30 g/day (6-15
g nitrogen/day).
• Urea constitutes 80% of urinary organic solids.

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 Metabolic Significant Aspects of Urea Cycle

A) Energy Cost:. Energy cost of the cycle is only one ATP.

B) urea cycle is related to TCA cycle:
1. CO2
2.Aspartate arises via transamination of oxaloacetate with
glutamate. Thus, depletion of oxaloacetate will decrease urea
formation (as in malonate poisoning).
3. Fumarate enters TCA cycle
C) Sources of Nitrogen in urea :free NH and
aspartate. 3

N.B. glutamate is the immediate source of both NH3 (via oxidative

deamination by Glu. Dehyd.) and aspartate nitrogen (through
transamination of oxaloacetate by AST).
 Importance of Urea Cycle

1. Formation of arginine (in organisms

synthesizing arginine) & formation of urea (in
ureotelic organisms, man) due to presence of
arginase.
2. Liver shows much higher activity of arginase
than brain or kidney for formation of urea while
in brain or kidney is the synthesis of arginine.
3. Synthesis of non-protein amino acids (ornithine
and citrulline) in body.
 Regulation of Urea Cycle
Activity of individual enzymes:
THE RATE LIMITING STEPS a) carbamoyl phosphate synthase-1
b) Ornithine transcarbamyolase.
c) Arginase.

N-acetylglutamate is activator for carbamoyl phosphate synthase-1

It enhances its affinity for ATP.
It is synthesized from acetyl CoA and glutamate.
its hepatic concentration increases after intake
of a protein diet, leading to an increased rate of urea synthesis.

is limited by the
Activity of ornithine transcarbamyolase
concentration of its co-substrate "ornithine".
 Change in the level of Enzymes:
Arginase & other urea-forming enzymes are adaptive enzymes
thus
a protein-rich diet will increase their biosynthesis rate &
the opposite is true for low protein diet.
However, in starvation, where the body is forced to use its own tissue
protein as fuel, there is an increase in urea-forming enzymes.

•
 ONE-CARBON FRAGMENT
METABOLISM
• Human body is unable to synthesize the methyl group and obtain it from diet.
• The first: met = S-adenosyl methionine (methyl donner = CH3 ) involved in
transmethylation reaction
• The second: tetrahydrofolic acid (FH or THF) which is a carrier of active
4

one-carbon units (-CH3, -CH2, -CHO, -CHNH, -CH).

 10 NH
N NADP+ NH NADP
CH 2 NADPH + H
+
NADPH + H+
6C N
5 9
C CH 2
8 7 CH
N DHF reductase DHF reductase
CH 2 10
N
(Folic acid) NH
5 H
H CH 2
N CH
(THF)
CH 2
N
H
(active form)
The one-carbon group carried by THF is attached to its N5 or N10 or
to both.
Group Position on THF
Methyl: -CH3 N5
Methylene -CH2 N5 , N10
Formyl - N5 or N10
CHO N5
Formimino N5 , N10
-CHNH

Methenyl -CH
• These one-carbon units are interconvertible to each other.
• The primary sources of one-carbon units are serine, glycine, histidine,
tryptophan & betaine.
• and their acceptors for biosynthesis a variety of biomolecules are
Phosphatidylethanolamine, Guanidoacetic acid, nor-Epinephrine ,Thymine,
Purine-C8 & Purine-C2 & homocysteine.
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