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By
SIR Moazzam Ali Shahid
 The complex of physical and chemical
processes occurring within a living cell or
organism that are necessary for the
maintenance of life.
 In metabolism some substances are broken
down to yield energy for vital processes while
other substances, necessary for life, are
synthesized.
 1. Occurs in the cytosol outside the
mitochondria.
 2. The breakdown of glucose to 2 pyruvate
molecules.
 1. Glycolysis begins with addition of 2
phosphate groups, activating glucose to react.
 2. Two separate reactions use 2 ATP.
 3. Glucose, a C6 molecule, splits into two C3
molecules, each with a phosphate group.
 Substrate Level Phosphorylation
Steps In Glycolysis
1. Hexokinase
Glucose + ATP  glucose-6-P + ADP
2. Phosphoglucose Isomerase catalyzes:
glucose-6-P (aldose)  fructose-6-P (ketose)
H O
O H
H
O HH
O H
CH 2O PO 3
2
H
O H
H
1
6
5
4
3 2
CH 2O PO 3
2
O H
CH 2O H
H
O H H
H HO
O
6
5
4 3
2
1
glucose-6-phosphate fructose-6-phosphate
Phosphoglucose Isom erase
3. Phosphofructokinase catalyzes:
Fructose-6-P + ATP  fructose-1,6-bisP + ADP
CH 2O PO 3
2
O H
CH 2O H
H
O H H
H HO
O
6
5
4 3
2
1 CH 2O PO 3
2
O H
CH 2O PO 3
2
H
O H H
H HO
O
6
5
4 3
2
1
ATP ADP
Mg
2+
fructose-6-phosphate fructose-1,6-bisphosphate
Phosphofructokinase
4. Aldolase catalyzes:
fructose-1,6-bisphosphate 
dihydroxyacetone-P + glyceraldehyde-3-P
.
6
5
4
3
2
1CH 2O PO 3
2
C
C
C
C
CH 2O PO 3
2
O
HO H
H O H
H O H
3
2
1
CH 2O PO 3
2
C
CH 2O H
O
C
C
CH 2O PO 3
2
H O
H O H+
1
2
3
fructose-1,6-
bisphosphate
Aldolase
dihydroxyacetone glyceraldehyde-3-
phosphate phosphate
Triosephosphate Isom erase
5. Triose Phosphate Isomerase (TIM) catalyzes:
dihydroxyacetone-P  glyceraldehyde-3-P
6
5
4
3
2
1CH 2O PO 3
2
C
C
C
C
CH 2O PO 3
2
O
HO H
H O H
H O H
3
2
1
CH 2O PO 3
2
C
CH 2O H
O
C
C
CH 2O PO 3
2
H O
H O H+
1
2
3
fructose-1,6-
bisphosphate
Aldolase
dihydroxyacetone glyceraldehyde -3-
phosphate phosphate
Triosephosphate Isom erase
C
C
CH 2O H
O O
H O PO 3
2
2
3
1
C
C
CH 2O PO 3
2
O O
H O H
2
3
1
3-phosphoglycerate 2-phosphoglycerate
Phosphoglycerate M utase
C
C
CH2OPO3
2
O O
H OPO3
2
2
3
1
2,3-bisphosphoglycerate
An active site histidine side-chain
participates in Pi transfer, by
donating & accepting phosphate.
The process involves a
2,3-bisphosphate intermediate.
9. Enolase catalyzes:
2-phosphoglycerate  phosphoenolpyruvate + H2O
C
C
C H 2O H
O O
H O PO 3
2
C
C
C H 2O H
O O
O PO 3
2
C
C
C H 2
O O
O PO 3
2
O H
2
3
1
2
3
1
H
2-phosphoglycerate enolate intermediate phosphoenolpyruvate
Enolase
10. Pyruvate Kinase catalyzes:
phosphoenolpyruvate + ADP  pyruvate +
ATP
C
C
CH 3
O O
O
2
3
1
A D P A T P
C
C
CH 2
O O
O PO 3
2
2
3
1
p h o sp h o en o lp yru vate p yru vate
P yru v ate K in ase
C
C
CH 3
O O
O
2
3
1
A D P A T PC
C
CH 2
O O
O PO 3
2
2
3
1
C
C
CH 2
O O
O H
2
3
1
phosphoenolpyruvate enolpyruvate pyruvate
Pyruvate K inase
Rate Limiting Enzyme
Controlled
EnzymeHexokinase
Phosphofructokinase
glucose Glycolysis
ATP
ADP
glucose-6-phosphate
Phosphoglucose Isomerase
fructose-6-phosphate
ATP
ADP
fructose-1,6-bisphosphate
Aldolase
glyceraldehyde-3-phosphate + dihydroxyacetone-phosphate
Triosephosphate
Isomerase
Glycolysis continued
Controlled
Enzyme
Rate Limiting
Enzyme
Glyceraldehyde-3-phosphate
Dehydrogenase
Phosphoglycerate K inase
Enolase
Pyruvate K inase
glyceraldehyde-3-phosphate
NAD
+
+ Pi
NADH + H
+
1,3-bisphosphoglycerate
ADP
ATP
3-phosphoglycerate
Phosphoglycerate M utase
2-phosphoglycerate
H2O
phosphoenolpyruvate
ADP
ATP
pyruvate
controlled
Enzyme
ATP Formation
Hexokinase
Phosphofructokinase
glucose Glycolysis
ATP
ADP
glucose-6-phosphate
Phosphoglucose Isomerase
fructose-6-phosphate
ATP
ADP
fructose-1,6-bisphosphate
Aldolase
glyceraldehyde-3-phosphate + dihydroxyacetone-phosphate
Triosephosphate
Isomerase
Glycolysis continued
-1
-1
Glyceraldehyde-3-phosphate
Dehydrogenase
Phosphoglycerate K inase
Enolase
Pyruvate K inase
glyceraldehyde-3-phosphate
NAD
+
+ Pi
NADH + H
+
1,3-bisphosphoglycerate
ADP
ATP
3-phosphoglycerate
Phosphoglycerate M utase
2-phosphoglycerate
H2O
phosphoenolpyruvate
ADP
ATP
pyruvate
Glyceraldehyde-3-phosphate
Dehydrogenase
Phosphoglycerate Kinase
Enolase
Pyruvate Kinase
glyceraldehyde-3-phosphate
NAD
+
+ Pi
NADH + H
+
1,3-bisphosphoglycerate
ADP
ATP
3-phosphoglycerate
Phosphoglycerate M utase
2-phosphoglycerate
H2O
phosphoenolpyruvate
ADP
ATP
pyruvate
3 ATP
Glucose + 2 ADP + 2 NAD+ + 2 Pi ---
--
> 2 Pyruvate + 2 ATP + 2 NADH + 2
H+
ATP utilized: 2
ATP produced: 4
NADH produced:2X3= 6
Total : 10- 2 = 8
Energy (Aerobic)
Glucose
2 Pyruvate
2 Ethanol 2 Acetyl-CoA 2 Lactate
4 CO 2 + 4 H 2O
Glycolysis
2 CO 2 –O 2 –O 2
+O 2
+O 2
CO 2
(yeast) (muscle)
What are the possible fates of pyruvate?
•Ethanol (fermentation)
•Acetyl coA (mammals and others)
•TCA/Krebs cycle
•Lactate (mammals and others)
•End product of anaerobic glycolysis
•Gluconeogenesis in liver via the Cori cycle
 In the presence of oxygen pyruvate is transported
into the mitochondria where it enters the next
major metabolic pathway for the production of
ATP energy.
 If there is no oxygen present then the pyruvate is
converted to a substance called lactate
Gibbs Free Energy Changes
Rxn# Enzyme
1 Hexokinase
2 Phosphogluco-isomerase
3 Phosphofructokinase
4 Aldolase
5 Triose phos. Isomerase
6 G-3-PDH
7 Phosphoglycerate kinase
8 Phosphoglycerate mutas
9 Enolase
10 Pyruvate kinase
11
2
3
10
9
8
7
6
5
4
References :
• Kaplan , Harpers Review,
• Internet

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Glycolysis by moazzam

Editor's Notes

  1. 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++.
  2. The mechanism involves acid/base catalysis, with ring opening, isomerization via an enediolate intermediate, and then ring closure. A similar reaction catalyzed by TriosephosphateIsomerase will be presented in detail.
  3. This highly spontaneous reaction has a mechanism similar to that of Hexokinase. The Phosphofructokinase reaction is the rate-limiting step of Glycolysis.
  4. The reaction is an aldol cleavage, the reverse of an aldol condensation.
  5. Glycolysis continues from glyceraldehyde-3-P. TIM's Keq favors dihydroxyacetone-P. Removal of glyceraldehyde-3-P by a subsequent spontaneous reaction allows throughput
  6. This dehydration reaction is Mg++-dependent. 2 Mg++ ions interact with oxygen atoms of the substrate carboxyl group at the active site.The Mg++ ions help to stabilize the enolate anion intermediate that forms when a Lys extracts H+ from C #2.
  7. This phosphate transfer from PEP to ADP is spontaneous. PEP has a larger DG of phosphate hydrolysis than ATP.Removal of Pi from PEP yields an unstable enol, which spontaneously converts to the keto form of pyruvate. Required inorganic cations K+ and Mg++ bind to anionic residues at the active site of PyruvateKinase.