Glycolysis

Dbanda
Mulungushi University

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 What is Glycolysis?
 It is a cascade of reactions that converts
glucose into two pyruvate molecules or into
lactate aiming at production of ATP and
other intermediates.
 It is also utilized in its opposite direction in
gluconeogenesis.

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 About GLYCOLYSIS

 D-glucose is the major fuel of most organisms

 D-glucose is rich in potential energy; completely catabolise to
CO2 and water proceeds with a standard free energy change
of –2,840 kJ/mol
 Glucose is also a versatile precursor capable of supplying
intermediates for biosynthesis
 Occurs in the cytosol of cells and is essentially an anaerobic
process
 Glycolysis occurs in the cytosol of cells and is essentially an
anaerobic process
 Role of Glycolysis is to produce energy and supply
intermediates for biosynthetic pathways 3
 Intracellular site and tissue
distribution:
It occurs in the cell cytosol of all tissues of the body.

1.RBCs:
are devoid of mitochondria and depend on glycolysis as the main source
of energy. Mammalian erythrocyte is unique in that about 90% of its total
energy requirement is provided by glycolysis.

2.Contracting muscles
due to occlusion of blood vessels by the muscular contraction that
decreases oxygen

3-Cornea, lens and some parts of retina

which have a limited blood supply and lack mitochondria which if present
would absorb and scatter light interfering with transparency. 4
4-Kidney (medulla), testicles, leukocytes
and white muscle fibers,
where there are relatively few mitochondria.

5-Cancer cells
due to dissociation of the high rate of
glycolysis from Krebs', i.e., aerobic production
of lactate.

6-Brain and gastrointestinal tract also

normally derive most of their energy from
glycolysis.
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 Biological importance (or
Functions) of glycolysis:
1. Glucose oxidation producing ATP.
2. It is the major source of energy in certain tissues, e.g.,
RBCs and skeletal muscles.
3. It provides pyruvic acid needed for Krebs' cycle.
4. It is a link with fat metabolism, e.g., dihydroxyacetone
phosphate into glycerol 3-phosphate in adipose tissue.
5. It a link with amino acid metabolism, e.g., 3-
phopshoglycerate into serine and pyruvate into alanine
and vice versa.
6. Production of 2,3-DPG that is important in tissue
oxygenation.

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 Other functions
7. It is the major source of lactic acid that is gluconeogenic.
8. Reversal of glycolysis is gluconeogenesis, an important
source of glucose.
9. Main pathway of metabolism of fructose from the diet.
10. A small number of genetic diseases occur due to deficiency
in activity of enzymes of glycolysis, are manifested mainly
as hemolytic anemias.
11. Cancer cells are glycolytic producing large amount of
lactate, favoring a relatively acidic local pH in the tumor, a
situation that was utilized to develop therapy for cancer
that could be locally activated by this acidic pH.

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 This pathway can be thought of as comprising three stages
STAGE I
 conversion of glucose into fructose 1,6-bisphosphate; consists of
three steps:
(i) A phosphorylation
(ii) An isomerisation, and
(iii) a second phosphorylation reaction
STAGE II
 Cleavage of fructose 1,6-bisphosphate into two three-carbon
fragments. The resulting three-carbon units are interconvertible
STAGE III
 ATP is harvested when the three-carbon fragments are oxidised
to pyruvate
The break down of the six-carbon glucose into two
molecules of the three carbon pyruvate occurs in ten
steps

The first 5 constitute the preparatory phase in which

two molecules of ATP are invested and the hexose
chain is cleaved into two triose phosphates

The last five constitute the pay-off phase, which

includes the energy-conserving phosphorylation steps
in which some of the free energy of the glucose is
conserved in the form of ATP

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The preparatory phase of glycolysis
1. Phosphorylation of Glucose

Glucose + ATP Glucose 6-phosphate + ADP+H+

 The reaction is irreversible

 Catalysed by hexokinase. Kinases are enzymes that catalyse

the transfer of a phosphoryl group from ATP to an acceptor

 Like many other kinases, hexokinase requires Mg 2+ for activity

 Hepatocytes contain hexokinase D or glucokinase which differs

from other forms of hexokinase in kinetics and regulatory
properties
2. Conversion of Glucose 6-Phosphate to Fructose 6-Phosphate
Glucose 6-Phosphate Fructose 6-Phosphate
 Catalyzed by Phosphohexose isomerase (phosphoglucose isomerase)
 Reaction is reversible
 Enzyme requires Mg 2+

3. Phosphorylation of Fructose 6-phosphate to fructose 1, 6-

bisphosphate ATP ADP
Fructose 6-phosphate Fructose 1,6-
bisphosphate

 Catalyzed by phosphofructokinase
 Reaction is irreversible
 Reaction is the major point of regulation in glycolysis
 Enzyme requires Mg2+
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4. Cleavage of Fructose 1,6-bisphosphate

Aldolase

Glyceraldehyde Dihydroxyacetone
3-phosphate Triose phosphate phosphate
isomerase
5.

 Dihydroxyacetone phosphate is rapidly and reversibly converted to glyceraldehyde

3-phosphate by triose phosphate isomerase
 4 and 5 are reversible reactions

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6. Oxidation of glyceraldehyde 3-phosphate to 1, 3-bisphosphoglycerate
NAD+ NADH
Glyceraldehyde 3-phosphate + Pi 1,3-bisphosphoglycerate
 Catalyzed by glyceraldehyde 3-phosphate dehydrogenase
 The first of the two energy-conserving reactions of glycolysis that eventually leads
to the formation of ATP
 Reversible
7. Phosphoryl transfer from 1, 3-bisphosphoglycerate to ADP to form ATP
ADP ATP
1,3-bisphosphoglycerate 3-phosphoglycerate

 The formation of ATP by phosphoryl group transfer from a substrate such as 1, 3

bisphosphoglycerate is referred to as substrate-level phosphorylation
 Reversible reaction but shifted towards products formation
 Catalyzed by phosphoglycerate kinase
 Requires Mg 2+

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8. Conversion of 3-phosphoglycerate to 2-phosphoglycerate
3-phosphoglycerate 2-phosphoglycerate
 Catalyzed by phosphoglycerate mutase
 Reversible

9. Dehydration of 2-phosphoglycerate to phosphoenolpyruvate

H2O
2-phosphoglycerate phosphoenolpyruvate
 Reversible
 Catalyzed by enolase. Inhibited by fluoride

10. Transfer of a phosphoryl group from phosphoenolpyruvate to ADP

ADP ATP
Phosphoenolpyruvate Pyruvate
 Irreversible reaction
 Catalyzed by pyruvate kinase
 Enzyme requires K+, Mg 2+ or Mn 2+
 Second substrate-level phosphorylation stage in glycolysis
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  From glucose to pyruvate there is a net production of 2 ATP molecules
 In the absence of oxygen pyruvate is reduced to lactate. This regenerates NAD+
that is required for the oxidation of glyceraldehyde 3-phosphate
 In the presence of oxygen it enters the mitochondria for further oxidation

 Tissues that function under hypoxic circumstances tend to produce

lactate
 True of skeletal muscles where the rate of work output may exceed the rate at
which oxygen is taken up and utilized. To enable glycolysis to continue
pyruvate is reduced to lactate generating NAD+

 In RBCs glycolysis even under aerobic conditions always terminates in lactate

because the subsequent reactions of pyruvate are mitochondrial and RBCs
lack mitochondria
 Other tissues that normally derive much of their energy from glycolysis and
produce lactate include brain, GIT, renal medulla, retina and skin
 Lactate in the liver is converted to glucose

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Control of glycolysis
 Enzymes catalyzing irreversible reactions are potential sites of
control ie hexokinase, phosphofructokinase and pyruvate
kinase
Phosphofructokinase
 Most important control site in mammalian glycolytic pathway
 In the liver the
 enzyme is inhibited by ATP
 Inhibitory effect of ATP is reversed by AMP
 Activity increases when the ATP/AMP ratio is lowered
 Inhibited by a drop in pH
 Inhibited by citrate
 Activated by Fructose 2,3-bisphosphate

Hexokinase
 Inhibited by glucose 6-phosphate
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Pyruvate kinase
Activated by fructose 1,6-bisphosphate, to
enable it to keep pace with the oncoming
high flux of intermediates
Inhibited by ATP
Inhibited by alanine
In the liver inhibited by phosphorylation

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 Feeder pathways for glycolysis
D-Galactose
Lactose Gylcogen;starch
Lactase
Phosphorylase

Glucose 1-phosphate
D-Glucose Phosphohexose
(gluco)mutase)
Glucose 6-phosphate D-Mannose
Sucrose
Sucrase

D-Fructose Fructose 6-phosphate

Fructose 1, 6-bisphosphate

Glyceraldehyde 3-bisphosphate
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Bioenergetics of (or Energy yield
from) glycolysis:

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Under anaerobic conditions:
1- Total ATP lost = 2 ATP as follows,
One ATP in the activation of glucose to glucose-6-phosphate.
One ATP in the activation of fructose-6-phosphate to fructose1,6-
diphosphate.

2- Total ATP gained = 4 ATP as follows,

2 ATP by substrate level phosphorylation from 1,3-
diphosphoglycerate
2 ATP from substrate level phosphorylation from phosphoenol
pyruvate.
3- Net ATP gained = 4 ATP gained - 2 ATP lost = 2 ATP for the
anaerobic oxidation of one mole of glucose into lactate.

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Under aerobic conditions:
Total ATP lost = 2 ATP.
Total ATP gained = 10 ATP are generated as follows,
4 ATP (obtained by substrate level phosphorylation) + 2
NADH.H+ chain (produced from oxidation of
glyceraldehyde-3-phosphate)  2 X 3 ATP = 6 ATP,
after oxidation in the functioning respiratory
Net ATP gained = 8 ATP as follows,
10 ATP – 2 ATP = 8 ATP for the aerobic oxidation of
one mole of glucose.

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Regulation (or Control) of
Glycolysis

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 A. Key regulatory enzymes:

are those enzymes that catalyze the irreversible

steps of glycolysis that include three steps as
follows,

1-Phosphofructokinase:
It is an allosteric enzyme stimulated by high levels of
fructose-6- phosphate, fructose-2,6-diphosphate (in
liver), ADP and AMP, Pi, and ammonia.
It is inhibited allosterically by ATP, low pH and citrate.

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2-Hexokinase:
Accumulation of glucose-6-phosphate and inhibition
of phosphofructokinase results in accumulation of
fructose-6-phosphate and glucose-6-phosphate that
allosterically inhibit hexokinase.
3-Pyruvate kinase: It is inhibited also by excess ATP,
fatty acids, and acetyl-CoA
and is stimulated by fructose-1,6-diphosphate, ADP
and AMP
It is regulated by cAMP-dependent phosphorylation-
dephosphorylation mechanism 26
 B. Hormonal regulation:

1. Insulin:
Stimulates synthesis of glucokinase,
phosphofructokinase and pyruvate kinase, so
it stimulates glycolysis.
It also induces glucose transporters to provide
cells with glucose for glycolysis.

2-Adrenaline and glucagon are

inhibitory by inhibiting pyruvate kinase. 27
END OF LECTURE

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