CHEM 160

GLYCOLYSIS
 CHEM 160
GLYCOLYSIS
Objectives
• Understand the metabolic processes that
carbohydrates undergo
• Describe the digestion and absorption of
carbohydrates
• Discuss the enzyme, co-factors, regulation and
significance of the glycolytic pathway
CARBOHYDRATE
DIGESTION
 Digestion of Carbohydrates

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Digestion of Carbohydrates

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THE GLYCOLYTIC
PATHWAY
Cellular Respiration: Recall
• Oxidation of food molecules to produce energy
(in the form of ATP) through a series of
cytoplasmic and mitochondrial-linked enzymatic
pathways.
• Consumes O2 and produces CO2 and H2O
Cellular Respiration: Recall
1. Glycolysis
2. Krebs Cycle
3. Electron Transport Chain
4. Synthesis of ATP
Glycolysis
• From the terms glycose (old name for glucose)
and lysis (degradation)
• A series of reactions that converts glucose to
pyruvate with accompanying production of ATP
and NADH.
• Location in the cell: CYTOSOL
• Also called Glycolytic Pathway or Embden-
Meyerhoff Pathway
Major Functions of Glycolysis
• To allow extraction of energy and conserve it in
the form of ATP
• To generate important biosynthetic precursors

ATP

+
alanine
Two Phases of Glycolysis
• Energy Investment Phase (Reactions 1 to 5)
• Glucose (6C) is cleaved to give 2 molecules of
glyceraldehyde-3-phosphate (3C each)
• Uses 2 ATP molecules
• Energy Pay-off Phase (Reactions 6 to 10)
• 2 glyceraldehye-3-phosphate molecules are
converted to 2 pyruvate molecules
• Generates 4 ATP and 2 NADH

• Yield: 2 ATP and 2 NADH

1. Hexokinase

• First regulatory point of glycolysis

Why phosphorylate glucose?
• Formation of
glucose-6-phosphate
increases the
concentration of
negative (−) charges
in the cytosol.
• Glucose is trapped
inside the cell and is
bpserrano 2009
activated for
breakdown
Glucose to Glucose-6-P
• Can be catalyzed by the ff. enzymes depending
on the tissue location and condition:
• Glucokinase (liver) – controls blood glucose
level; active at high [blood glucose]
• Hexokinase (muscle) – can be subjected to
allosteric inhibition by ATP
• Hexokinase (brain) – NO allosteric inhibition
by ATP
• Isozymes – enzymes that catalyze the same
reaction but differ in kinetic properties
2. Phosphoglucose isomerase
3. Phosphofructokinase (PFK-1)

• The COMMITTED STEP in glycolysis

• First irreversible reaction that is unique to
glycolysis
• Second regulatory point in glycolysis
4. Aldolase

• Sugar-splitting (“glyco-lysis”)
5. Triose Phosphate Isomerase

• End of energy investment phase

Summary of First Phase
• Glucose is cleaved to give
two moles of
glyceraldehyde-3-
phosphate
• 2 ATPs are consumed
6. Glyceraldehye-3-P dh
2 2

(2 moles) (2 moles) (2 moles)

• Uses an inorganic phosphate (Pi) to produce

1,3-BPG
• Note the coefficient (2)
• Two moles of NADH is produced in this step
7. Phosphoglycerate kinase
2 A D P 2 A T P

M g 2 +

2 p h o s p h o g ly c e r a t e
2
k in a s e

• Generation of 2 moles of ATP via substrate level

phosphorylation
8. Phosphoglyceromutase

2 2

• Shift of phosphate group from C-3 to C-2

9. Enolase
2

2 2

• Removal of two moles of H2O and formation of

a carbon-carbon double bond (dehydration)
10. Pyruvate kinase
2 ADP 2 ATP

2 Mg2+ 2
pyruvate kinase

• Generation of another two moles of ATP via

substrate level phosphorylation
• Third regulatory point in glycolysis
Summary of Second Phase
• 2 moles of glyceraldyde-3-
phosphate are converted
to 2 moles of pyruvate
• Produced in this phase
are:
• 4 moles of ATP
• 2 moles of NADH
• 2 moles of H2O
Overall Reaction:
First phase:
• Glucose + 2 ATP → 2 glyceraldehyde-3-P +
2 ADP

Second phase:
• 2 Glyceraldeyde-3-P + 4 ADP + 2 Pi + 2 NAD+ →
2 pyruvate + 4 ATP + 2 H2O + 2 NADH, H+

Net Reaction:
• Glucose + 2 ADP + 2 Pi + 2 NAD+ + → 2 pyruvate
+ 2 ATP + 2 H2O + 2 NADH, H+
Entry of Other Monosaccharides
Fructose
C H 2O H ATP ADP + Pi C H 2O P O 32-
C O C O
HO H HO H
H OH H OH
f r u c t o k in a s e
H OH H OH
C H 2O H C H 2O H
fru cto se fru c to se -1 -p h o sp h a te
Entry of Other Monosaccharides
Fructose
CH2O PO32- tri
ose
CH2O PO32- C O iso phos
me ph
C O CH2OH ras ate O H
e C
HO H dihydroxyacetone
fructose -1-P phosphate H OH
H OH
aldolase CH2O PO32-
H OH
nase glyceraldehyde-3-
CH2OH O H ki
C io se phosphate
t r
fructose-1-phosphate H OH ADP
CH2OH ATP
glyceraldehyde
Entry of Other Monosaccharides
Mannose
A
TP A
DP 2-
C
H2OH C
H2OP
O 3
O O
O
H O
H O
H O
H
h
exo
kin
ase
O
H O
H O H OH
α-D-mannopyranose-6-
α-D-mannopyranose
phosphate
ate
2- p
s h
CHOP
2 O3 C
H OH h
p ose
O 2
ose era
HO
anisom
n
H
O OH m
H O
α-D-fructofuranose-6-phosphate
Entry of Other Monosaccharides
Galactose
ATP ADP
CH2OH CH2OH
OH O OH O
OH galactokinase OH
OH O PO32-
OH OH
α-D-galactopyranose-1-
α-D-galactopyranose
phosphate
Entry of Other Monosaccharides
C H 2O H
OH O C H 2O H
O

OH O NH
OH g a la c t o s e 1 -P u r id y l O O
OH N O
O PO3 2- tra n sfe ra se O P O P O
OH O O O
OH
g a la c t o s e -1 -P OH OH
U D P -g a la c t o s e
O
C H 2O H t o se
O NH
a la c s e C H 2O H
OH O O N O D P -g m e ra O
OH O P O P O U epi
OH O O O
OH
U D P -g lu c o s e OH OH C H 2O P O 32- OH O P O 32-
c o OH
O g lu
s p ho g lu c o s e - 1 - P
OH o
p h ta se
u
OH OH m
g lu c o s e -6 -P O H
Galactosemia
• Caused by deficiency in galactose-1-P uridyl
transferase
• Results to high concentrations of galactose in
the blood
aldose reductase
Galactose Galactitol

• Accumulation of galactitol in the eyes causes

galactosemic cataract
Entry of Other Monosaccharides
REGULATION OF
GLYCOLYSIS
Why Regulate?
• To ensure that the output of the metabolic
pathways meet biological demands
• To ensure that energy in the form of ATP is not
wasted by having opposite pathways run
concomitantly in the same cell
Regulation of Glycolysis
• Regulatory points:
• Phosphofructokinase-1
• Hexokinase
• Pyruvate kinase
Phosphofructokinase-1
PFK−1
Fructose−6−P Fructose−1,6−bP
• Main regulatory point of glycolysis
• Inhibitors:
ATP - When energy charge is high
- ATP allosterically inhibits
PFK-1
- no glucose should enter
glycolysis
- glucose is stored as glycogen
Phosphofructokinase-1
• Inhibitors:
citrate
high [citrate]
means that
there are
enough
substrates
(pyruvate and
acetyl-CoA)
present
Phosphofructokinase-1
• Activators:
ADP/AMP
• high cellular concentrations of these
metabolites signal a low energy state
• There is a need to metabolize glucose
for energy
Phosphofructokinase-1
• Activators:
Fructose-2,6-bisphosphate
PFK−2
• Fructose-6-P Fructose-2,6-bP
• Formed when [Fructose-6-P] is high
• Reduces the inhibitory effect of ATP on
PFK-1 and enhances binding of fructose-
6-P to PFK-1
Hexokinase
hexokinase
Glucose Glucose−6−P

• Glucose-6-P inhibits hexokinase via feedback

inhibition
• Enzyme action is inhibited by its product(s)
• If PFK-1 is inhibited, Fructose-6-P accumulates
as well as Glucose-6-P
• ∴ hexokinase is subsequently inhibited
Pyruvate Kinase
PK
phosphoenolpyruvate + ADP pyruvate + ATP

• Inhibitors via feedback inhibition:

• ATP
• Alanine
• Acetyl CoA
• Signals that building blocks are abundant
Pyruvate Kinase
PK
phosphoenolpyruvate + ADP pyruvate + ATP

• Inhibitor via covalent modification:

low blood glucose level
• Triggers a cascade of signals that lead to
phosphorylation of pyruvate kinase,
rendering it inactive
• Activator: Fructose-1,6-bisphosphate
FATES OF PYRUVATE
Fates of Pyruvate
Fermentation
Alcoholic fermentation in yeast and other
microorganisms
Fermentation
Lactic acid fermentation in vertebrates
Lactate accumulation
• During strenuous exercise, cells become
depleted with O2 supply → oxygen debt
• Aerobic respiration is slowed down and
pyruvate accumulates
• Pyruvate is instead converted to lactate which,
upon accumulation, lowers intracellular pH
causing painful muscle cramps
Lactose Intolerance
• Deficiency in the enzyme lactase
lactase
lactose glucose + galactose

• Lactose is a good source of energy for

microorganisms in the colon

fermentation
lactose lactic acid + H2 +CO2 +CH4
References
CHEM 160 lecture slide of Profs. BP Serrano, KMP Caldo and AC Reyes

Nelson and Cox. Lehninger’s Principles of Biochemistry, 4th ed.

Stryer, Berg and Tymoczko. Biochemistry 4th ed.