Glycolysis

Shekhar Chandra Yadav

Lecturer
Dept. of Biochemistry
 Glycolysis
Definition:
• Glycolysis is the sequence of 10 reactions that break
down one molecule of glucose to:
– two molecules of pyruvate (3C) (in presence of O2) or to
– 2 molecules of lactic acid (in absence of O2).
– with net generation of 2 molecules of - ATP and NADH

Location:
– All cells of the body.

Site:
– In the cytoplasm of every cell,
 Importance Of Glycolysis
1) Supply energy
– 2 ATP in absence of O2 and
– 8 ATP in the presence of O2

2) Supply 2 molecules of Pyruvic acid which enter the

mitochondria and converted into:
– Acetyl-CoA (active acetate) that are oxidized by
Citric Acid Cycle
– Oxaloacetic acid, which is the initiator of citric
acid cycle.

3) Supply Dihydroxy Acetone Phosphate which is

converted to α­Glycerophosphate, which can be
converted to Triglyceride (Lipogenesis)
4)Synthesis of amino acids:
from intermediates of glycolysis e.g.
– 3-phosphoglycerate Serine

– Pyruvic acid alanine

5)Importance of Glycolysis in RBCs

– Energy production:
• It is the only pathway that supplies the
red cells with 2 ATP.
– Reduction of met-hemoglobin:
• Glycolysis provides NADH for reduction
of met­Hb in red cells
 The Individual Reactions of Glycolysis

The pathway consisting of 3 separate phases.

• The first require energy:(Energy Investment Phase)

– (2 molecules of ATP are used to convert glucose to fructose 1,6-
bisphosphate (F1,6 BP)

• The second is splitting phase

• The third is the energy-producing phase:

– F1,6BP is degraded to 2 molecules of pyruvate, with the
production of 4 molecules of ATP and 2 molecules of NADH]
 1)The Hexokinase(glucokinase) Reaction

• This reaction consumes 1 molecule of ATP, and is catalyzed by

enzyme hexokinases/glucokinase.

• It is irreversible reaction.

• Glucokinase is an isoform of hexokinase enzyme present in liver.

• It requires Mg-ATP complex as substrate. Un-complexed ATP is a

potent competitive inhibitor of this enzyme.

• This enzyme undergoes large conformational change upon

binding with Glucose. It is inhibited allosterically by G6P.
2) Phosphohexose Isomerase reaction:
Glucose 6- Phosphate isomerase Fructose-6-Phosphate

 This enzyme catalyzes the reversible

isomerization of G6P (an aldohexose) to F6P
(a ketohexose).

 This enzyme requires Mg ++ for its activity.

 It is specific for G6P and F6P.

 3) Phosphofructokinase, (PFK)

Fructose 6- Phosphate Phosphofructokinase fructose 1,6-bisphosphate

ATP ADP

• This reaction is irreversible.

• This is the most important step in regulation of glycolysis.
• The enzyme Phosphofructokinase is one of the most
complex regulatory enzymes, with various allosteric
inhibitors and activators.
• ATP is an allosteric inhibitor
• ADP and AMP also activate PFK whereas citrate is an
inhibitor.
 4) Aldolase

fructose 1,6-bisphosphate Aldolase dihydroxyacetone phosphate + glyceraldehyde 3-

P

5) Triose Phosphate Isomerase:

dihydroxyacetone phosphate Isomerase glyceraldehyde 3-P

 6) Glyceraldehyde-3-Phosphate Dehydrogenase
Glyceraldehyde-3-Phosphate Dehydrogenase 1,3- bisphosphoglycerate

Pi NAD+ NADH + H+
• This reaction is reversible.
 
• Fate of NADH produced:
– In aerobic glycolysis, NADH is oxidized in respiratory chain
resulting in synthesis of 3 ATP molecules
– In anaerobic glycolysis, NADH is oxidized in reaction catalyzed
by lactate dehydrogenase.
 7) Phosphoglycerate Kinase
1,3-bisphosphoglycerate Kinase 3-phosphoglycerate
ADP ATP
• The high-energy phosphate of 1,3-BPG is used to form ATP
(substrate level phosphorylation)
• This reaction is reversible.
N.B.
• In erythrocytes, there is an associated pathway in which 1,3
biphosphoglycerate is converted to 2,3BPG
• 2,3 BPG is an important regulator of hemoglobin's affinity for
oxygen. Note that 2,3-bisphosphoglycerate phosphatase degrades
2,3BPG to 3-phosphoglycerate, a normal intermediate of glycolysis
without production of ATP.

1,3-bisphosphoglycerate mutase 2,3-bisphosphoglycerate

2,3-bisphosphoglycerate phosphatase 3- phosphoglycerate
8) Phosphoglycerate Mutase:
3-phosphoglycerate mutase 2- phosphoglycerate
– This reaction is reversible
9) Enolase
2- phosphoglycerate Enolase phosphoenolpyruvate

10) Pyruvate Kinase

• phosphoenoylpyruvate pyruvate kinase pyruvic acid
ADP ATP
• In this reaction, ATP is formed (substrate level phosphorylation)
• This reaction is irreversible
 Lactate dehydrogenase
• This reaction occurs only in anaerobic
glycolysis (in RBCs and exercising muscles)

• It oxidizes NADH produced by glyceraldehydes

3 P dehydrogenase enzyme by reducing
pyruvate to lactate.
The Energy derived from glycolysis
• Glycolysis requires two molecules of ATP in the reactions catalyzed by
Hexokinase, Phosphofructokinase

• Then four molecules of ATP are formed by substrate level

phosphorylation in the reactions catalyzed by:
– phosphoglycerate kinase,
– pyruvate kinase.

• Thus anaerobic glycolysis produce 2 molecules of ATP.

• Aerobic glycolysis produce, in addition, two molecules of

NADH which are transported to mitochondria to produce 6
ATP molecules by oxidation via respiratory chain. The net
result is 8 ATP.
 Inhibitors of glycolysis
• Glyceraldehyde 3P dehydrogenase enzyme is inhibited
by arsenic which form 1-arseno 3-phosphoglycerate,
thus prevent formation of ATP by phosphoglycerate
kinase enzyme.

• Enolase enzyme is inhibited by floride. Thus floride is

used during collection of blood samples for estimation
of glucose, to inhibit glycolysis to keep blood glucose
 Regulation of Glycolysis:
• Two types controls for metabolic reactions:

• a) Substrate limited : When concentrations of reactant and products in the

cell are near equilibrium, then it is the availability of substrate which
decides the rate of reaction.

• b) Enzyme-limited: When concentration of substrate and products are far

away from the equilibrium, then it is activity of enzyme that decides the
rate of reaction. These reactions are the one which control the flux of the
overall pathway.

• There are three steps in glycolysis that have enzymes which regulate the
flux of glycolysis.

I. The hexokinase (HK)

II. The phoshofructokinase (PFK)
III.The pyruvate kinase
• Hexokinase: It is allosterically inhibited by its product Glucose 6
phosphate. In liver Glucokinase is inhibited by Fructose 6
Phosphate. Uncomplexed ATP acts as a competitive inhibitor of this
enzyme.

• Phosphofructokinase (PFK):
• It’s activity is controlled by a complex allosteric regulation.
• This reaction commits the cells to channel glucose to glycolysis.
• ATP is the end product of glycolysis as well as it is substrate for PFK.
In presence of high concentration of ATP, ATP binds to inhibition site
of PFK, and thereby decreases the activity of enzyme.
• AMP, ADP and Fructose 2, 6 biphosphate act as allosteric activators
of this enzyme.
• Activation of enzyme by AMP overcomes the inhibitory effect of
ATP.
• Pyruvate Kinase: It is allosterically inhibited by ATP. ATP
binding to the inhibitor site of pyruvate kinase decreases its
ability to bind to phosphoenol pyruvate (PEP) the substrate.
• It is also inhibited by Acetyl coenzyme A and long chain fatty
acid.
 Fructose
ATP
Hexokinase
ADP
Fructose-6-phosphate

Galactose
ATP Galactokinase
ADP
Galactose-1-phosphate
Galactose-1-phosphate uridyl
transferase
Glucose-1-phosphate

phosphoglucomutase

Glucose-6-phosphate
 Disease associated with Glycolysis
• Genetic disease- Pyurvate Kinase Deficiency-
Hemolytic Anemia
• Cancer – Glycolysis increases 200
times( Warburg Effect)
• Lactic Acidosis- Increased Lactate in tissues,
Low blood PH
THANK YOU