Glycolysis

Embden Meyerhof pathway

o Gustav Embden
o Otto Meyerhof

1922 Nobel Prize

 Glycolysis
(glyco = sugar; lysis = to break
down)

Where do reactions of glycolysis

occur in the cell?

Where does the glucose (or other

sugar) come from?

Do these reactions need oxygen to

generate energy?

What is the end-product of

glycolysis?

Figure 4-13
o The glycolytic pathway is working in the cytosol of

all tissues for the breakdown (oxidation) of glucose.

o To provide energy (in the form of ATP) and

intermediates for other metabolic pathways.

o It can work in the presence of O2 (aerobic

glycolysis), and in the absence of O2 (anaerobic

glycolysis).
 Aerobic glycolysis

o Pyruvate is the end product of glycolysis in cells

with mitochondria and an adequate supply of

oxygen.

o Oxygen is required to reoxidize the NADH formed

during the Aerobic glycolysis, through the ETC.

o Hydrogen of NADH enters the mitochondria by

glycerol phosphate shuttle (producing 2 ATPs) or

by malate shuttle (producing 3ATPs)

 Anaerobic glycolysis

o Allows the continued production of ATP in

tissues that lack mitochondria (for example, red

blood cells) or in cells deprived of sufficient

oxygen (cotracting muscles).

o Pyruvate, is reduced by NADH to form lactate,

this allows regeneration of oxidized NAD for

further reaction in the absence of O2 .

REACTIONS OF GLYCOLYSIS
The 1st investment of
1
an ATP in glycolysis.

CHO CHO
I I
HCOH HCOH
I Glucokinase I
HOCH(1) Preparatory phase
Hexokinase HOCH
I I
HCOH HCOH
I I
HCOH ATP ADP HCOH
I I
CH2OH CH2O - P

Glucose Glucose, 6-
phosphate

1st Reaction: Glucose enters cell and is phosphorylated to glucose 6-

phosphate, which is trapped inside the cell.
 2

CHO CH2OH
I I
HCOH C=O
I Phosphoglucose I
HOCH isomerase HOCH
I I
HCOH HCOH
I I
HCOH HCOH
I I
CH2O - P CH2O - P

Glucose, 6- Fructose, 6-
phosphate phosphate

2nd Reaction: the isomerization of glucose 6-phosphate to fructose 6-

phosphate. Catalyzed by phosphoglucose isomerase.
 The 2nd investment of
an ATP in glycolysis. 3

CH2OH CH2O- P
I I
C=O C=O
I Phosphofructokinase-1 I
HOCH HOCH
I I
HCOH HCOH
I I
HCOH ATP ADP HCOH
I I
CH2O - P CH2O - P

Fructose, 6- Fructose 1,6 -

phosphate bisphosphate

Bis means two phosphate groups on two different carbon atoms. Di means
two phosphate groups linked together on the same carbon atom.
 4 CH2O- P
I
C=O
I
CH2O- P CH2OH
I
C=O
I Dihydroxyacetone
HOCH Aldolase phosphate
I
HCOH
I
HCOH HC=O
I I
CH2O - P HCOH
I
CH2O - P
Fructose 1,6 -
bisphosphate Glyceraldehyde-
3 - phosphate

Reverse aldol condensation; converts a 6 carbon atom sugar to 2

molecules, each containing 3 carbon atoms.
 5

Triose
CH2O- P phosphate HC=O
I isomerase I
C=O HCOH
I I
CH2OH CH2O - P

Dihydroxyacetone Glyceraldehyde-
phosphate 3 - phosphate
 6

O
HC=O II
(2) Payoff phase. CO~ P
I
HCOH I
I HCOH
CH2O - P I
NAD+ + Pi NADH + H CH2O - P
Glyceraldehyde, 1,3-bisphospho
3-phosphate Glycerate
Substrate-level
phosphorylation 7

O
II
CO - P Phosphoglycerate HOC=O
I kinase I
HCOH HCOH
I I
CH2O - P CH2O - P
ADP ATP

1,3- bisphospho 3- phospho

Glycerate Glycerate

1st example of substrate level phosphorylation.

The net yield from glycolysis is 2 ATP
 8

HOC=O phosphoglyceromutase HOC=O

I I
HCOH HCO - P
I I
CH2O - P CH2OH

3- phospho 2- phospho
Glycerate Glycerate

The phosphoryl group is shifted from the C-3 to the C-2 position of
glycerate. Note: A mutase transfers a functional group from one position
to another on the same molecule.
 9

HOC=O HOC=O
I Enolase I
HCO- P CO- P
I II
CH2OH CH2
H2O
2- phospho phospho
Glycerate enolpyruvate

Dehydration reaction
Substrate-level 10
phosphorylation

Pyruvate
HOC=O kinase HOC=O
I I
CO- P C=O
I I
CH2 CH3
ADP ATP

phospho Pyruvate
enolpyruvate

2nd example of substrate level phosphorylation.

The net yield from glycolysis is 2 ATP
 Glucose
ATP
1 Glucokinase
ADP
Hexokinase
Glucose 6-phosphate

2 Phosphoglucose
isomerase
Fructose 6-phosphate
(1) PreparatoryATP
phase
3 Phosphofructokinase
ADP

Fructose 1,6-bisphosphate

4 Aldolase

Dihydroxyacetone Glyceradehyde 3-
phosphate phosphate
Triose phosphate
5 isomerase
 Glyceraldehyde 3-phosphate
NAD+ + Pi
Glyceraldehyde 3-
6 phosphate dehydrogenase NADH + H+
1,3-Bisphosphoglycerate
ADP
7 Phosphoglycerate kinase
ATP
(2) Payoff phase.
3-Phosphoglycerate
8 Phosphoglyceromutase
2-Phosphoglycerate

9 Enolase
H2O

Phosphoenolpyruvate
ADP
10 Pyruvate kinase
ATP

pyruvate
 Glucose
Aerobic ATP

glycolysis ATP

o Glucose 
2 pyruvate
+ 2 ATP
+ 2 NADH NADH
NADH

ATP ATP

ATP
ATP

Pyruvate Pyruvate
 Anaerobic Glucose
glycolysis ATP

o Glucose  2 ATP

lactate + 2 ATP

NADH + H+ NADH + H+

NAD+ NAD+

ATP ATP

ATP ATP

Lactate Pyruvate Pyruvate Lactate

 NAD+ NADH + H+

Glyceral- Glyceraldehyde 3-P 3-Phosphoglyceric acid

dehyde 3-P (enzyme-bound) (enzyme-bound)
Pi

Free Enzyme

1,3-Bisphosphoglyceric acid
 Glucose
Aerobic glycolysis ATP

ATP

• Glucose 
2 pyruvate
+ 2 ATP
+ 2 NADH
NADH
NADH

ATP ATP

ATP
ATP

Pyruvate Pyruvate
 Glucose
Anaerobic glycolysis
ATP

• Glucose  ATP

2 Lactate
+ 2 ATP

NADH + H+ NADH + H+

NAD+ NAD+

ATP ATP

ATP ATP

Lactate Pyruvate Pyruvate Lactate

I. Energy production
• Under aerobic conditions
• Under anaerobic conditions
II. Importance of glycolytic intermediates
1. Pyruvate
• Oxaloacetate
• Acetyl CoA
2. Dihydroxyacetone phosphate (DHAP)
3. Synthesis of nonessential amino acids
III. Importance of glycolysis in red blood cells
• Energy production
• Production of methemoglobin
• Synthesis of 2,3-bisphosphoglycerate (2,3 BPG)
IV. Gluconeogenesis
I- Energy production.

• Under aerobic conditions

glucose  2 pyruvate + 6 or 8 ATP.

(according to shuttle used)

• Under anaerobic conditions

glucose  2 lactate + 2 ATP.

 II- Importance of glycolytic intermediates:

1. Pyruvate

• Pyruvate  oxaloacetate (the prime of Krebs’ cycle)

• Pyruvate  acetyl CoA

• Oxidized in Krebs’ cycle for production of energy

• Supply the building unit for FA and cholesterol

synthesis.
2. Dihydroxy acetone phosphate (DHAP)

• DHAP  glycerol 3-phosphate (for synthesis of

Triglyceride and phospholipids).

3. Synthesis of non-essential amino acids glycine

cysteine

• 3- phosphoglycerate  serine

• Pyruvate  alanine
III- Importance of glycolysis in red blood cells

• Energy production

• Reduction of methemoglobin by NADH-cyt b5-

methemoglobin reductase

• Synthesis of 2,3 bisphosphoglycerate (2,3 BPG)

which units with hemoglobin at low oxygen tension
and helps release of O2 to the tissues.
Glyceraldehyde 3-phosphate

1,3 bisphosphoglycerate
Bisphosphoglycerate
mutase
ADP
Mg2+
ATP
Pi H2O
3- phosphoglycerate 2,3 BPG
2,3 BPG phosphatase

Pyruvate
Deficiency of glycolytic enzymes, e.g. pyruvate kinase
deficiency (PK):

• PK deficiency is restricted to RBCs and

produces mild to severe chronic hemolytic anemia

depending on the degree of enzyme deficiency.

 The enzyme may
show an abnormal
Km or Vmx for
substrates or
coenzymes.

Phosphoenolpyruvate

ADP
Pyruvate Fructose 1,6-
kinase + bisphosphate
ATP
The enzyme may
Enzyme activity or Pyruvate show an abnormal
stability may be response to the
altered, or the activator fructose
amount of enzyme Lactate 1,6-bisphosphate.
may be decreased.
IV- Reversal of glycolysis (gluconeogenesis)

• During fasting and low dietary carbohydrates

glycolysis is reversed for synthesis of glucose from

non-carbohydrate compounds (lactate, glycerol and

amino acids). All the reactions are reversed by the

same enzymes except the three irreversible kinases.

Regulation of glycolysis
Glucose
Glucagon
- -
Inhibition - HK GK
+ Insulin
+ Stimulation

Glucose-6- P
ATP
Citrate
- Frucotse-6- P
- Glucagon
AMP PFK-
Fructose-6-P + 1 + Insulin
Fructose-1,6-BP
Fructose-2,6-BP

Glyceraldehyde-3-P

Phosphoenolpyruvate
- Glucagon
ATP + PK
- Insulin
+
Pyruvate
 Effect of hormones on glycolysis
1. Insulin

• induces synthesis of the 3 irreversible enzymes.

• Dephosphorylates the enzymes through activation

of protein phosphatase.

2. Glucagon

• Represses (inhibit) the synthesis of the 3

irreversible enzymes.

• Phosphorylates the enzymes mediated by cAMP

activation of protein kinase.
1- Regulation of hexokinase and glucokinase

Hexokinase Glucokinase
site extrahepatic Liver and pancreatic -cells
Substrate All hexoses Glucose mainly
Enzyme kinetics • High affinity (low Km) • Low affinity (high Km) and it
and it is active at low is only active at high glucose
glucose concentration, concentration. So these cells
which ensure supply of metabolize glucose only when
glucose for tissues it is elevated.
• Low Vmax • High Vmax
Glucose-6- Allosteric inhibitor No effect
phosphate

Effect of
• Insulin • No effect • Inducer (transcription of GK
• Glucagon • No effect gene)
• Fasting • No effect • Repressor
• Feeding of • No effect • Decreases activity
carbohydrates • Increases activity
2- Regulation of phosphofructokinase-1 (PFK-1)

• regulation by the energy levels within the cell PFK-1 allosterically

inhibited by ATP and citrate.

• allosterically activated by AMP and fructose 2,6-bisphosphate.

 Glucagon

Adenylate
cyclase
ATP cAMP

Active protein kinase A Fructose 2,6-

Fructose 6- bisphosphate
phosphate
P P
ATP ADP
Phosphofructo
kinase-1 PFK-2 FBP-2 PFK-2 FBP-2

Fructose 1,6-
bisphosphate
Fructose 6-
phosphate

Glycolysis

Fasting = inhibition of glycolysis in the liver

Insulin

Activation of many enzymes

Fructose 6- Fructose 6-
phosphate phosphate P P
Phosphofructo
PFK-2 FBP-2 PFK-2 FBP-2
kinase

Fructose 1,6-
bisphosphate

Fructose 2,6-
bisphosphate

Glycolysis

Feeding = activation of glycolysis in the liver

3- regulation of pyruvate kinase (PK)

A. Feed-forward regulation by fructose 1,6-bisphosphate.

B. Covalent modification of pyruvate kinase.

 Glucagon
Receptor

Adenyl
cyclase
ATP cAMP + PPi

Active protein kinase A

ADP PEP
P
PEP ATP ADP
Pyruvate Pyruvate
kinase kinase
(active) (inactive)
pyruvate Pi H2O

Protein pyruvate
+ phosphatase ATP
Insulin
 Glucagon

R C

R C cAMP 2 C
Inactive protein Active protein
+ 2{ R -2 cAMP}
kinase A kinase A

ATP ADP

Dephosphorylated Phosphorylated
P
protein protein
Mg2+
Pi H2O
Protein
- PK a (active) phosphatase - PK b (inactive)

- PFK-2a (active) - PFK-2b (inactive)

- F-2,6-Bpase b (inactive) - F-2,6-BPase a (active)

Activation of Insulin Inhibition of

glycolysis glycolysis

Insulin action Glucagon action

Summary
 Glucose
Aerobic glycolysis ATP

ATP

• Glucose 
2 pyruvate
+ 2 ATP
+ 2 NADH
NADH
NADH

ATP ATP

ATP
ATP

Pyruvate Pyruvate
 Glucose
Anaerobic glycolysis
ATP

• Glucose  ATP

2 Lactate
+ 2 ATP

NADH + H+ NADH + H+

NAD+ NAD+

ATP ATP

ATP ATP

Lactate Pyruvate Pyruvate Lactate

Regulation of glycolysis
Glucose
Glucagon
- -
Inhibition - HK GK
+ Insulin
+ Stimulation

Glucose-6- P
ATP
Citrate
- Frucotse-6- P
- Glucagon
AMP PFK-
Fructose-6-P + 1 + Insulin
Fructose-1,6-BP
Fructose-2,6-BP

Glyceraldehyde-3-P

Phosphoenolpyruvate
- Glucagon
ATP + PK
- Insulin
+
Pyruvate