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unit-1

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prasenjitbhattacharjee2010
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© © All Rights Reserved
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BRIEF STUDY OF BASIC METABOLIC PATHWAY AND

FORMATION OF DIFFERENT SECONDARY


METABOLITES THROUGH VARIOUS PATHWAY
•The sum total of all the enzymatic mediated reactions occurring in the cell is collectively
called metabolism.
•The reaction sequences occurring within organisms in an orderly and regulated way are
known as metabolic pathways.
•The compounds formed during metabolism are called metabolites.

Cellular metabolism has four functions


1. To obtain chemical energy i.e ATP through degradation of energy reach biomolecules

2. To transform biomolecules into building blocks or precursors needed for the synthesis of
macromolecular cell components

3. To assemble building blocks into proteins, nucleic acids, lipids and other cell components

4. To form and degrade biomolecules required in the specialized functions of cells


BASIC METABOLIC PATHWAY
BASIC METABOLIC PATHWAY
 The most important building blocks used in the biosynthesis of secondary

metabolites are derived from the intermediates acetyl coenzyme A (acetyl-CoA),

shikimic acid, mevalonic acid, and 1 deoxyxylulose 5 phosphate.

 These are utilized respectively in the acetate, shikimate, mevalonate, and deoxy

xylulose phosphate pathways.


BASIC METABOLIC PATHWAY
SHIKIMIC ACID PATHWAY
 Metabolic pathway for biosynthesis of aromatic amino acid(phenylalanine,
tyrosine, tryptophane)
 Shikimic acid is key intermediate from carbohydrate for biosynthesis of C 6-C3 unit
(phenyl propane derivative). Ex-Tyrosine and phenyl alanine
 Shikimic acid name is derived from shikimi, Japense flower (star anise, Ilicium
anisatum)
 Occur in plant, bacteria, fungi but not in animal.
 BIOSYNTHETIC PRECURSOR -

(starting material)
 Phosphoenol pyruvate (gycolysis)
 Erythrose- 4- phosphate (pentose phosphate pathway)
 Nitrogen from other amino acid – glutamate, glycine, serine
 Phenylalanine and tyrosine form the basis of C6-C3 phenylpropane units found in
many natural products, for example, cinnamic acids, coumarins, lignans, and
flavonoids, and along with tryptophan are precursors of a wide range of alkaloid
structures.
GLYCOLYSIS
ACETATE PATHWAYS
ACETATE PATHWAYS
•Imp metabolic pathway for synthesis of secondary metabolites.
•As starting material utilizes in this pathway is Acetate (acetic acid) from Glycolysis.
•This acetate is utilize in the form of Acetyl CoA, as it is the active form of acetate.
A. ACETATE-MELONATE PATHWAY:
Precursor: Acetyl CoA (From Glycolysis)
Intermediate: Malonyl CoA
End products: Fatty Acids Polyketides
Aromatic
(Saturated Compounds
& Unsaturated Fatty Acids)

Polyketides
Pathway

Polyacetylenes
Glycosides Secondary
Phospholipids metabolites
Glycolipids
Waxes
B. ACETATE-MEVALONATE PATHWAY:

•Also known as Isoprenoid pathway.

•Starting Material: Acetate (Acetyl CoA).

•One of the intermediate formed is Mevalonate or mevalonic acid.

•This mevalonic acid further used for the synthesis of different types of isoprenoids.

•These isoprenoids are utilised for the synthesis of terpenes and steroids.

•So in different parts of different plants / in specific cells of different plants,

different terpenes (like mono, sesqui, di, tri, poly) and steroids are biosynthesized

by Acetate mevalonate pathway,


AMINO ACID PATHWAY
 Plants and bacteria can synthesize all 20 of the amino acids. Whereas

humans cannot synthesize 9 of them.


 These 9 amino acids must come from our diets and are called essential

acids. EX- Histidine, Isoleucine, Leucine, Lysine, Methionine,


Phenylalanine, Threonine, Tryptophan, and Valine.
 The 11 amino acids are called non-essential amino acids like Alanine,

Arginine, Aspargine, Aspartate, Cysteine, Glutamate, Glutamine,


Glycine, Proline, Serine and Tyrosine.
 The non-essential amino acids are synthesized by simple pathways,

whereas biosynthesis of the essential amino acids are complex.


 All 3 aromatic amino acids are derived from shikimate pathway.
STUDY OF UTILIZATION OF RADIOACTIVE
ISOTOPES IN THE INVESTIGATION OF
BIOGENETIC STUDIES

There are 5 techniques used for the investigation


of biosynthetic pathway of primary and secondary
metabolites-
1. Tracer technique,
2. Use of isolated organ and tissues,
3. Grafting method,
4. Use of Mutant strains,
5. Enzymatic studies.
Out of the above 5 methods, in Tracer technique
method radioactive isotopes are used for the
investigation of biogenetic studies.
TRACER TECHNIQUE
 Significance/Importances of tracer techniques-
 Applicable for living systems. Wide ranges of isotopes are
available.
 High sensitivity
 More effective
 Simple administration and isolation.
 Shows accurate results when enough metabolic time & technique
is used.
 Position & Quantity of compound containing tracer isotope 14C
marked glucose is used for glucose determination in the biological
system.
 For different studies, different tracers can be used. For studies on
nitrogen and amino acid, Labelled nitrogen gives specific
information than carbon.
 Biosynthetic pathway can be traced by incorporating radioactive
isotopes into the precursor or starting material. e.g- By
incorporation of 14C to phenyl alanine, the biosynthesis of
Cyanogenetic glycosides, Prunacin can be traced. Location and
quantity can be determined in biological system.
TRACER TECHNIQUE

Different trace elements used for different


studies-
1. For studies on protein, alkaloids and amino
acid, nitrogen atom gives more specific
information than carbon.
2. For studies on glycosidic linkage- O, N, S
and C atom.
3. For studies on terpenoids- O atom.
TRACER TECHNIQUE
Steps involved in tracer techniques-
1. Preparation of labelled compound,

2. Incorporation of labelled compound,

3. Separation and isolation of labelled

compound,
4. Determination of nature of metabolites in

various biochemical fractions.


TRACER TECHNIQUE
 1. Preparation of labelled compound-
 In biological investigation, the use of bioactive isotopes enables the metabolism of
compounds to be followed in living organisms for detection and estimation of soft
and easily absorbed radiation from labelled compound.
 Labelled compounds may be prepared by use of radioactive isotopes and stable
isotopes. e,g- Radioactive isotopes- C, 3H,
14
P,
32
I
131

 Stable isotopes- 2H, 15


N, C,
13
O
18

 Radioactive carbon and hydrogen are mostly used in biological investigation.


 Radioactive isotopes having long half-life are used.

Criteria for selection of trace elements-


 Starting concentration of trace element must be sufficient to withstand dilution in
the course of metabolism.
 Physical and chemical nature of compound must be known.
 Half-life should be sufficiently long.
 Should not damage the tissue system
 Should have low radiation energy.
 Instruments used to detect properties of metabolites are Scintillation chamber,
GM counter, Autoradiography, NMR and MS- ionization technique.

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