CARBOHYDRATE METABOLISM

GYCOLYSIS

Chapter Outline
Features of glycolysis, coupled reactions,
chemical principles
Features of the 1st and 2nd phase of glycolysis

Major pathways of carbohydrate

metabolism.
Energy source

Structural
elements in living
cell

Overview of Glycolysis

The Embden-Meyerhof (Warburg) Pathway

Essentially all cells carry out glycolysis
Occurs throughout the cytoplasm (is not organelle
associated)
Ten reactions - same in all cells - but rates differ
Two phases:
First phase converts glucose to two G-3-P (4 steps)
Second phase produces two pyruvates (6 steps)
Products are pyruvate, ATP and NADH
Three possible fates for pyruvate

Overview of Glycolysis
Occurs without the use of oxygen (anerobic
metabolism; fermentation)
Energy is transferred in several steps to ATP
The overall glucose lytic reaction:
Glucose + 2 NAD+ + 2 ADP + 2 Pi 2 pyruvic
acid + 2 NADH + 2 ATP + 2H+ + 2 H2O
Pyruvic acid can continue to ferment into
lactic acid

10 Reactions of glycolytic pathway

1. Synthesis of
glucose-6phosphate

2. Conversion of
G6P to F6P

8. Interconversion
of 3phosphoglycerate
and 2phosphoglycerate

7. Phosphoryl
group transfer

9. Dehydration of
2phosphoglycerate

10. Synthesis of
pyruvate

3. Phosphorylation
of F6P

4. Cleavage of F1,6-B

6. Oxidation of
glyceraldehyde-3phosphate

5. Interconversion
of glyceraldehyde3-phosphate and
dihydroxyacetone
phosphate

GLYCOLYSIS: FIRST
PHASE

Rx 1: Synthesis of glucose-6-phosphate
The first reaction phosphorylation of glucose
Hexokinase or glucokinase
This is a priming reaction ATP is consumed here in
order to get more later
ATP makes the
phosphorylation of glucose
spontaneous

Glycolysis: Step 1
glucose
CH2 OH
O H
H
H
OH H
HO
OH
H
OH

+ ATP
8P1-11

2+
hexokinase, Mg
2CH2 OPO3

O H

H
HO

H
OH
H

+ ADP

OH
OH

glucose-6-phosphate

Hexokinase
1st step in glycolysis; G large, negative
Hexokinase (and glucokinase) act to phosphorylate
glucose and keep it in the cell
Km for glucose is 0.1 mM; cell has 4 mM glucose
So hexokinase is normally active!
Glucokinase (Kmglucose = 10 mM) only turns on when cell
is rich in glucose
Hexokinase is regulated - allosterically inhibited by
(product) glucose-6-P - but is not the most important
site of regulation of glycolysis

Rx 2: Phosphoglucoisomerase
Glucose-6-P to Fructose-6-P
Why does this reaction occur??
next step (phosphorylation at C-1)
would be tough for hemiacetal OH, but easy for primary -OH

Aldose G6P converted to F6P by

phosphoglucose isomerase (PGI)
Ene-diol intermediate in this
reaction
Transformation make C1 of
fructose available for
phosphorylation

Glycolysis: Step 2
glucose-6-phosphate
2CH2 OPO3

O H

H
HO

H
OH
H

8P1-15

OH
OH

phosphoglucose
isomerase
2-

CH2O PO3
O CH2OH
H HO
H
OH
OH H

fructose-6-phosphate

Rx 3: Phosphorylation of fructose-6phosphate
Phosphofructokinase-1
(PFK-1)
irreversibly
catalyze
phosphorylation of F6P to F-1,6-B
After F-1,6-B synthesized, the cell
is committed to glycolysis
F1,6b split into 2 triose
PFK1-major regulatory enzyme in
glycolysis.
Its
activity
is
allosterically inhibited by high
level of ATP and citrate

Glycolysis: Step 3
fructose-6-phosphate
2-

CH2O PO3
phosphofructokinase-1
CH
OH
O
2
(PFK-1), Mg2+
H HO
2CH2O PO3
H
2OH
O CH2OPO3
OH H
H HO
+ ATP
+ ADP
H
OH
OH H
8P1-17

fructose-1,6-bisphosphate

Regulation by PFK-1
Enzyme
Hexokinase

Activator

Inhibitor
Glucose-6Phosphate, ATP

PFK-1

Fructose-2,6-bis Citrate, ATP

phosphate, AMP

Pyruvate
kinase

Fructose-1,6-bis Acetyl-CoA, ATP

phosphate, AMP

Rx 4: cleavage of Fructose-1,6Bisphosphate
C6 cleaves to 2 C3s (DHAP,
Gly-3-P)
Reaction involves: aldol cleavageproduct are aldehyde & ketone
End of stage 1 glycolysis
Although cleavage of F-1,6-B is
frequently unfavorable
(G=+23.8 kJ/mol), the reaction
proceed because the products are
rapidly removed

Rx 5: Triose Phosphate Isomerase

DHAP converted to Gly-3-P
Of the two products of the aldolase reaction,
only 3GP serves as substrate for next reaction in
glycolysis
To prevent loss of other 3C unit from glycolytic
pathway, triose phosphate isomerase catalyze
interconversion of DHAP and G3P
After this reaction, 2 molecules G3P are
produced

Glycolysis - Second Phase

Metabolic energy produces 4 ATP
Net ATP yield for glycolysis is two ATP
Second phase involves two very high energy
phosphate intermediates
1,3 BPG
Phosphoenolpyruvate

Rx 6: Oxidation of gly-3-p
Gly-3P is oxidized to 1,3-BPG
A phosphorylation and a two electron
oxidation by NAD+ occur.
G-3-P + NAD+ + H2O 3-P-glycerate
(- 43.1 kJ)
3-P-glycerate + HPO42- + H+ glycerate1,3-bisP
(+49.3 kJ)

O N
H2N C
S H HC O
B H C OH
2CH2OPO3

O N
H2N C
H
S
C O
B H H C OH
+
2CH2OPO3
O N
H2N C
H H
S
C O
B H H C OH
+
2CH2OPO3

NAD+

O N
H2N C
H H
S
C O
B H H C OH
+
2CH2OPO3

N
O
NADH
O
H2N C
H O P O
S
C
O
+
OH
H
H
B
H C OH
+
2CH2OPO3

Glycerate-1,3-bisphosphate
and original enzyme

Rx 7: Phosporyl group transfer

ATP synthesis from a high-energy
phosphate
ATP is synthesized as phosphoglycerate
kinase catalyze transfer of high energy
phosphoryl group of gly-1,3-b to ADP
This is referred to as "substrate-level
phosphorylation

Rx 8: Interconversion of 3-phosphoglycerate
and 2-phosphoglycerate

Phosphoryl group from C-3 to C-2

Rationale for this enzyme - repositions the
phosphate to make PEP
3-phosphoglycerate has a low phosphoryl
group transfer potential
It is a poor candidate for further ATP synthesis
Cells convert gly-3-p with its energy-poor
phosphate ester to phosphoenolpyruvate
(PEP)-high phosphoryl group transfer
potential

Rx 9: dehydration of 2phosphoglycerate
2-P-Gly to PEP
Enolase catalyze dehydration of gly-2-p to PEP
How can such a reaction create a PEP?
"Energy content" of 2-PG and PEP are
similar
Enolase just rearranges to a form from
which more energy can be released in
hydrolysis

Rx 10: synthesis of pyruvate

PEP to Pyruvate makes ATP

Catalyze by pyruvate kinase (transfer of phosphoryl
group from PEP to ADP)
These two ATP (from one glucose) can be viewed as
the "payoff" of glycolysis
Large, negative G - regulation!
Allosterically activated by AMP, F-1,6-bisP
Allosterically inhibited by ATP and acetyl-CoA
Understand the keto-enol equilibrium of Pyruvate

The Fate of NADH and Pyruvate

Aerobic or anaerobic??

NADH is energy - two possible fates:

If O2 is available, NADH is re-oxidized in the electron
transport pathway, making ATP in oxidative
phosphorylation
In anaerobic conditions, NADH is re-oxidized by lactate
dehydrogenase (LDH), providing additional NAD+ for
more glycolysis

The Fate of NADH and Pyruvate

Aerobic or anaerobic??

Pyruvate is also energy - two possible fates:

aerobic: citric acid cycle
anaerobic: LDH makes lactate

Other Substrates for Glycolysis

Fructose, mannose and galactose
Fructose and mannose are routed into
glycolysis by fairly conventional means.
Galactose is more interesting - the Leloir
pathway "converts" galactose to glucose - sort
of.

Substrate cycle
Occurs when two metabolic pathways run simultaneously in opposite
directions and have no overall effect other than to dissipate energy in the form
of heat.
A pair of non-reversible reactions that cycle between two substrates are called
as a substrate cycle
Level of substrate cycling is very minimal because of reciprocal regulation of
the enzymes
Certain organisms utilize such reactions to maintain body temperature

Identify the pairs of reaction in glycolysis which is known

as the substrate cycle.

REFERENCES
McKee T.M., McKee J.R. (2003). Biochemistry. The
Molecular Basis of Life, 4th Edition, McGraw Hill.
Garrett, R., & Grisham, C. (2010). Biochemistry, 4th
edition, Thomson Brooks/Cole.