Two phases of glycolysis

The conversion of glucose to pyruvate occurs in

two stages .The first five reactions of glycolysis
correspond to an energy investment phase in
which the phosphorylated forms of
intermediates are synthesized at the expense of
ATP.
The subsequent reactions of glycolysis constitute
an energy generation phase in which a net of
two molecules of ATP and 2 NADH are formed by
substrate-level phosphorylation per glucose
molecule metabolized.
REACTIONS OF GLYCOLYSIS
1. Phosphorylation of glucose
Phosphorylated sugar molecules do not readily
penetrate cell membranes, because there are no
specific transmembrane carriers for these
compounds, and because they are too polar to
diffuse through the lipid core of membranes.
Mammals have several isozymes of the enzyme
hexokinase that catalyze the phosphorylation of
glucose to glucose 6-phosphate.
Hexokinase has broad substrate specificity and is
able to phosphorylate several hexoses in addition
to glucose.
2. Isomerization of glucose 6-phosphate
The isomerization of glucose 6-phosphate to
fructose 6-phosphate is catalyzed by phospho
glucose isomerase
3. Phosphorylation of fructose 6-phosphate
The irreversible phosphorylation reaction
catalyzed by phospho - fructokinase-1 (PFK-1) is
the most important control point and the rate-
limiting and committed step of glycolysis PFK-1
is controlled by the available concentrations of
the substrates ATP and fructose 6- phosphate.
4. Cleavage of fructose 1,6-bisphosphate
Aldolase cleaves fructose 1,6-bisphosphate to
dihydroxy acetone phosphate and glyceraldehyde 3-
phosphate
5. Isomerization of dihydroxyacetone phosphate
Triose phosphate isomerase interconverts
dihydroxyacetone phosphate and glyceraldehyde 3-
phosphate. Dihydroxy - acetone phosphate must be
isomerized to glyceraldehyde 3-phosphate for further
metabolism by the glycolytic pathway. This
isomerization results in the net production of two
molecules of glycer - aldehyde 3-phosphate from the
cleavage products of fructose 1,6- bisphosphate.
 6. Oxidation of glyceraldehyde 3-phosphate
The conversion of glyceraldehyde 3-phosphate to 1,3-
bisphosphoglycerate by glyceraldehyde 3-phosphate
dehydrogenase is the first oxidation-reduction
reaction of glycolysis. Because there is only a limited
amount of NAD+ in the cell, the NADH formed by this
reaction must be reoxidized to NAD+ for glycolysis to
continue. Two major mechanisms for oxidizing NADH
are:
1) the NADH-linked conversion of pyruvate to lactate
(anaerobic)
2) oxidation of NADH via the respiratory chain
(aerobic)
7. Synthesis of 1,3-bisphosphoglycerate (1,3-
BPG):
The oxidation of the aldehyde group of
glyceraldehyde 3-phosphate to a carboxyl
group is coupled to the attachment of Pi to the
carboxyl group. The high-energy phosphate
group at carbon 1 of 1,3-BPG conserves much
of the free energy produced by the oxidation
of glyceraldehyde 3-phosphate. The energy of
this high-energy phosphate drives the
synthesis of ATP in the next reaction of
glycolysis.
Synthesis of 2,3-bisphosphoglycerate (2,3-
BPG) in red blood cells:
Some of the 1,3-BPG is converted to 2,3-BPG
by the action of bisphosphoglycerate mutase
(see Figure). 2,3-BPG, which is found in only
trace amounts in most cells, is present at high
concentration in red blood cells (increases O2
delivery) .2,3-BPG is hydrolyzed by a
phosphatase to 3-phosphoglycerate, which is
also an intermediate in glycolysis .
8. Synthesis of 3-phosphoglycerate producing ATP
When 1,3-BPG is converted to 3-
phosphoglycerate, the high-energy phosphate
group of 1,3-BPG is used to synthesize ATP from
ADP. This reaction is catalyzed by
phosphoglycerate kinase.Because two
molecules of 1,3-BPG are formed from each
glucose molecule, this kinase reaction replaces
the two ATP molecules consumed by the earlier
formation of glucose 6-phosphate and fructose
1,6-bisphosphate.
Shift of the phosphate group from carbon 3
to carbon 2
The shift of the phosphate group from carbon
3 to carbon 2 of phosphoglycerate by
phosphoglycerate mutase is freely reversible
9. Dehydration of 2-phosphoglycerate
The dehydration of 2-phosphoglycerate by
enolase redistributes the energy within the 2-
phosphoglycerate molecule, resulting in the
formation of phosphoenolpyruvate (PEP),
which contains a high energy enol phosphate.
10. Formation of pyruvate producing ATP
The conversion of PEP to pyruvate is catalyzed
by pyruvate kinase. The equilibrium of the
pyruvate kinase reaction favors the formation
of ATP
Reduction of pyruvate to lactate
Lactate, formed by the action of lactate
dehydrogenase, is the final product of
anaerobic glycolysis in eukaryotic cells.
The formation of lactate is the major fate for
pyruvate in lens and cornea of the eye, kidney
medulla, testes, leukocytes and red blood
cells, because these are all poorly vascularized
and/or lack mitochondria.
 Energy yield from glycolysis
• Despite production of ATP in glycolysis, the
end product still contains most of energy
contained in glucose. The TCA cycle I required
to release that energy completely.
• 1. Anaerobic glycolysis: two molecules of ATP
are generated for each molecule of glucose
converted to two molecules of lactate. There
is no net production or consumption of NADH.
 2. Aerobic glycolysis
• The direct consumption and formation of ATP is the
same as in anaerobic glycolysis (that is a net gain of
two ATP per molecule of glucose). Two molecules of
NADH are also produced per molecule of glucose.
Ongoing aerobic glycolysis requires the oxidation of
most of this NADH by the electron transport chain,
producing approximately three ATP for each NADH
molecule entering the chain (NADH cannot cross the
inner mitochondrial membrane, and substrate
shuttle are required.
Energy from glycolysis
• ATP consumed 2 moles
• ATP produced direct 4 moles
• ATP indirect (NADH/H) 6 moles
• Net ATPs = 10-2= 8 moles
• If anaerobic glycolysis 2 moles
Lactate formation in muscle:
In exercising skeletal muscle, NADH
production exceeds the oxidative capacity of
the respiratory chain. This results in an
elevated NADH/NAD+ ratio, favoring reduction
of pyruvate to lactate. Therefore, during
intense exercise, lactate accumulates in
muscle, causing a drop in the intracellular pH,
potentially resulting in cramps.
Lactic acidosis:
Elevated concentrations of lactate in the plasma,
termed lactic acidosis, occur when there is a
collapse of the circulatory system, such as in
myocardial infarction, pulmonary embolism, and
uncontrolled hemorrhage, or when an individual
is in shock. The failure to bring adequate
amounts of oxygen to the tissues results in
impaired oxidative phosphorylation and
decreased ATP synthesis. To survive, the cells use
anaerobic glycolysis as a backup system for
generating ATP, producing lactic acid as the end
product.
REGULATION OF GLYCOLYSIS
During the well-fed state:
Decreased levels of glucagon and elevated levels of
insulin, such as occur following a carbohydrate- rich
meal, cause an increase in fructose 2,6-bisphosphate
and, thus, in the rate of glycolysis in the Fructose 2,6-
bisphosphate, therefore, acts as an intracellular signal,
indicating that glucose is abundant.
During starvation:
Elevated levels of glucagon and low levels of insulin,
such as occur during fasting decrease the intracellular
concentration of hepatic fructose 2,6-bisphosphate.
This results in a decrease in the overall rate of
glycolysis and an increase in gluconeogenesis.
Glycolysis