MUHAMMAD WAJID
Institute of Pharmacy Gulab Devi Educational
Complex, Lahore
Metabolism:
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Metabolism is “the entire set of enzyme- catalyzed
transformations of organic molecules in living cells”
Two broad classes
Catabolism & Anabolism Catabolic Pathways
Transform fuels into cellular energy
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Conti…
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Requires inputs of energy to proceed
Useful energy + small molecules complex molecules
Pathways that can be either anabolic or catabolic,
depending on the energy conditions in the cell are
referred to as amphibolic pathways
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Glycolysis:
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Derived from Greek word Glykys (Sweet) and Lysis
(Splitting)
The process in the cell metabolism by which
carbohydrates and sugar, especially glucose, are
broken down, producing ATP and pyruvic acid and two
“high energy” electron carrying molecules of NADH
During this process one molecule of glucose (6 carbon
molecule) is degraded into two molecules of pyruvate
(3 carbon molecule)
It is also called as Embden-Mayerhof Parnas Pathway
or EMP pathway
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Conti…
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It was the first metabolic sequence to be studied
It take place in all body cells
It occurs in cytosol
The enzymes of this pathway are present in the
cytosomal fraction of the cell
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Introduction:
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Glycolysis is the sequence of 10 enzyme-catalyzed
reactions that converts glucose into pyruvate with
simultaneous production of ATP
In this oxidative process, 1molecule of glucose is
partially oxidized to 2 molecules of pyruvate
This major pathway of glucose metabolism occurs in
the cytosol of all cell
This unique pathway occurs aerobically as well as
anaerobically & doesn’t involve molecular oxygen
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Conti…
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The glycolytic sequence of reactions differ from
species to species only in the mechanism of its
regulation & in the subsequent metabolic fate of the
pyruvate formed
In aerobic organisms, glycolysis is the prelude to Citric
acid cycle and ETC
Glycolysis is the central pathway for Glucose
catabolism
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Conti…
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Glycolysis occurs in the absence of oxygen
(Anaerobic) or in the presence of oxygen (Aerobic)
Lactate is the end product under anaerobic
conditions
In aerobic condition, pyruvate is formed, which is
then oxidized to CO2 and H2O
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Two phases of glycolysis:
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Glycolysis leads to breakdown of 6-C glucose into
two molecules of 3-C pyruvate with the enzyme
catalyzed reactions being bifurcated or categorized
into 2 phases:
o Phase 1- preparatory phase
o Phase 2- payoff phase
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Preparatory phase:
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It consists of the 1st 5 steps of glycolysis in which the
glucose is enzymatically phosphorylated by ATP to
yield Fructose-1,6-bisphosphate
This fructuse-1,6-biphosphate is then split in half to
yield 2 molecules of 3-carbon containing
Glyceraldehyde-3-phosphate/ dihyroxyacteone
phosphate
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Conti…
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Thus the first phase results in cleavage of the
hexose chain
This cleavage requires an investment of 2 ATP
molecules to activate the glucose molecule and
prepare it for its cleavage into 3-carbon
compound
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Payoff phase:
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This phase constitutes the last 5 reactions of Glycolysis
This phase marks the release of ATP molecules during
conversion of Glyceraldehyde-3-phosphtae to 2
molecules of Pyruvate
Here 4 moles of ADP are phosphorylated to ATP.
Although 4 moles of ATP are formed, the net result is
only 2 moles of ATP per mole of Glucose oxidized,
since 2 moles of ATP are utilized in Phase 1
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Stepwise explanation of glycolysis:
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Phosphorylation:
Glucose is phosphorylated by ATP to form glucose-
6- phosphate
This is an irreversible reaction & is catalyzed by
hexokinase
6 CH2OH 6 CH OPO 2
2 3
ATP ADP
5 O 5 O
H H H H
H H
4 1 4 H 1
OH H 2+ OH
Mg
OH OH OH OH
3 2 3 2
H OH Hexokinase H OH
glucose glucose-6-phosphate
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Hexokinase:
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The reaction involves nucleophilic attack of the C6
hydroxyl O of glucose on P of the terminal phosphate
of ATP
ATP binds to the enzyme as a complex with Mg++
Mg++ interacts with negatively charged phosphate
oxygen atoms, providing charge compensation &
promoting a favorable conformation of ATP at the
active site of the Hexokinase enzyme
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Isomerization:
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It is a reversible rearrangement of chemical
structure of carbonyl oxygen from C1 to C2,
forming a Ketose from the Aldose
Thus, isomerization of the aldose Glucose 6-
phosphate gives the ketose, Fructose-6-phoshphate
6 CH OPO 2
2 3
5 6 CH OPO 2 1 CH2OH
H O H 2 3
O
H
4 H 1 5 H HO 2
OH
OH OH H 4 3 OH
3 2
OH H
H OH
Phosphoglucose Isomerase
glucose-6-phosphate fructose-6-phosphate
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Phosphorylation:
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Here the Fructose-6-phosphate is phosphorylated by
ATP to fructose-1,6-bisphosphate
This is an irreversible reaction and is catalyzed by
phosphofructokinase enzyme
Phosphofructokinase
6 CH OPO 2 1CH2OH 6 CH OPO 2 1CH2OPO32
2 3 2 3
O ATP ADP O
5 H HO 2 5 H HO 2
H 4 3 OH Mg2+ H 4 3 OH
OH H OH H
fructose-6-phosphate fructose-1,6-bisphosphate
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Breakdown:
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This six carbon sugar is cleaved to produce two 3-
C molecules: glyceradldehyde-3-phosphate (GAP)
& dihydroxyacetone phosphate(DHAP)
This reaction is catalyzed by Aldolase
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1 CH2OPO 3
2C O
H O
2
HO 3 C H Aldolase 3
CH2OPO3 1C
H 4C OH 2C O + H 2
C OH
2
H C OH 1CH2OH 3 CH2OPO 3
5
2
6 CH2OPO 3 dihydroxyacetone glyceraldehyde-3-
phosphate phosphate
fructose-1,6-
bisphosphate
Triosephosphate Isomerase
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Isomerization:
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Dihydroxyacetone phosphate is oxidized to form
Glyceraldehyde-3-phosphate
This reaction is catalyzed by triose phosphate
isomerase enzyme
Triosephosphate Isomerase
H H OH H
+
O
+ + +
H C OH H H C H H C
C O C OH H C OH
CH2OPO32 CH2OPO32 CH2OPO32
dihydroxyacetone enediol glyceraldehyde-
phosphate intermediate 3-phosphate
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Phosphorylation:
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This is the first reaction of energy yielding step.
Oxidation of aldehyde derives the formation of a
high energy acyl phosphate derivate
Glyceraldehyde-3-phosphate
Dehydrogenase
H O + H+ O OPO 32
1C NAD+ NADH 1 C
+ Pi
H C OH H C OH
2 2
2 2
3 CH2OPO 3 3 CH2OPO 3
glyceraldehyde- 1,3-bisphospho-
3-phosphate glycerate
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Conti….
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An inorganic phosphate is incorporated in this
reaction without any expense of ATP
NAD+ is the cofactor in this reaction which acts as an
oxidizing agent
The free energy released in the oxidation reaction is
used in the formation of acyl phosphate
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Dephosphorylation:
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This reaction generates ATP from the energy
released by the oxidation of 3-Phosphoglycerate
The enzyme undergoes substrate-induced
conformational change similar to that of Hexokinase
Phosphoglycerate Kinase
O OPO 32 ADP ATP O O
1C 1
C
H 2
C OH H 2
C OH
2+
2 Mg 2
3 CH2OPO 3 3 CH2OPO 3
1,3-bisphospho- 3-phosphoglycerate
glycerate
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Phosphoglycerate Mutase:
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Phosphate is shifted from the OH on C3 to the OH on
C2
Phosphoglycerate Mutase
O O O O
C
1
C
1
H 2C OH H 2C OPO32
2
3 CH2OPO3 3 CH2OH
3-phosphoglycerate 2-phosphoglycerate
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Enolase Reaction:
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Enolase
O H O OH O
O O O
C C 1
C
1
H 2 C OPO32 C OPO32 2C OPO32
3 CH2OH CH2OH 3 CH2
2-phosphoglycerate enolate intermediate phosphoenolpyruvate
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Pyruvate Kinase:
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Pyruvate Kinase
O O O O
ADP ATP
1
C 1
C
2
C OPO32 2
C O
3 CH2 3 CH3
phosphoenolpyruvate pyruvate
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Conti…
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glucose Glycolysis
ATP
Hexokinase
ADP
glucose-6-phosphate
Phosphoglucose Isomerase
fructose-6-phosphate
ATP
Phosphofructokinase
ADP
fructose-1,6-bisphosphate
Aldolase
glyceraldehyde-3-phosphate + dihydroxyacetone-phosphate
Triosephosphate
Isomerase
Glycolysis continued
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Conti….
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glyceraldehyde-3-phosphate
NAD+ + Pi Glyceraldehyde-3-phosphate
NADH + H + Dehydrogenase
1,3-bisphosphoglycerate
ADP
Phosphoglycerate Kinase
ATP
3-phosphoglycerate
Phosphoglycerate Mutase
2-phosphoglycerate
H2O Enolase
phosphoenolpyruvate
ADP
Pyruvate Kinase
ATP
pyruvate
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Regulation of glycolysis:
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The rate of glycolysis is regulated by controlling of the 3
irreversible enzymes (key enzymes)
These enzymes catalyze what is called committed reactions of the
pathway
These enzymes are
o glucokinase (hexokinase)
o phospho-fructokinase-1
o pyruvate kinase
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Hormonal regulation:
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Insulin: Stimulates synthesis of all key enzymes of
glycolysis
It is secreted after meal (in response to high blood
glucose level)
Glycogen:
Inhibits the activity of all key enzymes of glycolysis.
It is secreted in response to low blood glucose level
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Energy regulation:
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High level of ATP inhibits PFK-1 and pyruvate kinase
High level of ADP and AMP stimulate PFK-1
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Substrate regulation:
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Glucose-6-phosphate inhibits hexokinase (and not
glucokinase)
Fructose 2,6 bisphosphate stimulates
phosphofructokinase-1
Citrate inhibits phosphofructokinase-1
Fructose 1,6 bisphosphate stimulates pyruvate kinase
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Muhammad Wajid
