Lesson Learning Outcome

Upon completion of this lecture, students should

be able to:
understand the glycolytic pathway
fates of pyruvate
The Glycolytic Pathway
Glucose is converted to two pyruvate
Glycolysis
Glycolysis: a series of 10 enzyme-catalyzed
reactions by which glucose is oxidized to two
molecules of pyruvate
O
glycolysis
C6 H12 O6 2CH3 CCOO- + 2H+
Glucose Pyruvate

there is net conversion of 2ADP to 2ATP

O
C6 H12 O6 + 2ADP + 2Pi 2CH3 CCOO- + 2ATP
Glucose Pyruvate
 Fates of Pyruvate
Pyruvate is most commonly metabolized in
one of three ways, depending on the type of
organism and the presence or absence of O 2

aerobic conditions
plants and animals 3CO2 + 2H2 O

O OH
anaerobic conditions
CH3 CCOO- CH3 CHCOO-
contracting muscle
Pyruvate Lactate
anaerobic conditions
CH3 CH2 OH + CO2
fermentation in yeast
Ethanol
 Reactions of glycolysis
Reaction 1: phosphorylation of -D-glucose to give
glucose-6-phosphate
CH2 OH
HO O O O
HO hexokinase
+ -O-P-O-P-O-AMP 2+
OH Mg
O- O-
-D-Glucose OH ATP

CH2 OPO3 2-
HO O O
HO
+ -O-P-O-AMP
OH O-
OH
-D-Glucose-6-phosphate ADP
 this reaction is driven by the free energy of
hydrolysis of ATP

Glucose + P i Glucose-6-phosphate + H2 O Go' = +13.8 kJ•mol -1

ATP + H2 O ADP + Pi Go' = -30.5 kJ•mol -1

Glucose + ATP Glucose-6-phosphate + ADP Go' = -16.7 kJ•mol -1

The enzyme that catalyzes this reaction is

hexokinase
Glucose-6-phosphate inhibits hexokinase –
feedback inhibition
Reaction 2: isomerization of glucose-6-phosphate to
fructose-6-phosphate
6 6
CH2 OPO3 2- CH2 OPO3 2- 1
phosphogluco- CH2 OH
HO O isomerase O
HO H HO 2
2
OH
1 H OH
OH HO H
-D-Glucose-6-phosphate -D-Fructose-6-phosphate

The enzyme that catalyzes this reaction is

glucosephosphate isomerase
The aldehyde group at C1 is reduced to hydroxyl, and
the C2-hydroxyl is oxidized to give the ketone group
of fructose-6-phosphate
 Reaction 3: phosphorylation of fructose-6-phosphate
6
CH2 OPO3 2- 1
O CH2 OH phospho-
H HO fructokinase
+ ATP
H OH Mg 2+
HO H
6
-D-Fructose-6-phosphate CH2 OPO3 2- 1
O CH2 OPO3 2-
H HO + ADP
H OH
HO H
-D-Fructose-1,6-bisphosphate

The phosphorylation of fructose-6-phosphate is

highly exergonic and irreversible – enzyme
responsible is phosphofructokinase
Reaction 4: cleavage of fructose-1,6-bisphosphate to
two triose phosphates by enzyme aldolase
CH2 OPO3 2- CH2 OPO3 2-
C=O Dihydroxyacetone
C=O phosphate
aldolase CH2 OH
HO H
H OH CHO
H OH D-Glyceraldehyde
H C OH 3-phosphate
2-
CH2 OPO3
CH2 OPO3 2-
Fructose-1,6-bisphosphate
 Reaction 5: isomerization of triose phosphates
catalyzed by triosephosphate isomerase
reaction involves two successive keto-enol
tautomerizations
only the D enantiomer of glyceraldehyde 3-
phosphate is formed

CH2 OH CHOH CHO

C=O C-OH H C OH
CH2 OPO3 2- CH2 OPO3 2- CH2 OPO3 2-
Dihydroxyacetone An enediol D-Glyceraldehyde
phosphate intermediate 3-phosphate
Reaction 6: oxidation of the -CHO group of D-
glyceraldehyde-3-phosphate
the -CHO group is oxidized to a carboxyl group
the oxidizing agent, NAD+, is reduced to NADH

A two-electron oxidation
O O
G- C- H + H2 O G- C- OH + 2H+ + 2e-

A two-electron reduction
NAD+ + H+ + 2e- NADH

O O
G- C- H + H2 O + NAD+ G- C- OH + H+
 the overall reaction involves an exergonic oxidation and
an endergonic phosphorylation
 the overall reaction is slightly endergonic

O O
oxidation: C-H to C-O- Go' = -43.1 kJ•mol -1

O O O
phosphorylation: C-O- to C-O-P-O- Go' = +49.3 kJ•mol -1
O-

O O O
C-H to C-O-P-O- Go' = +6.2 kJ•mol -1
O-
Reaction 7: transfer of a phosphate group from 1,3-
bisphosphoglycerate to ADP
this reaction is called substrate-level phosphorylation

O
phospho-
C-OPO3 2- O glycerate kinase
H C OH + -O-P-O-AMP
CH2 OPO32- O- Mg2+
1,3-Bisphospho-
glycerate ADP
COO- O O
H C OH + -O-P-O-P-O-AMP
CH2 OPO32- O- O-
3-Phosphoglycerate ATP
 this reaction is the sum of the endergonic
phosphorylation of ADP and the exergonic hydrolysis of
the mixed phosphate anhydride
phosphorylation:
ADP + Pi ATP + H2 O Go' = +0.5 kJ•mol -1
hydrolysis:
O O O
C-O-P-O- + H2 O C-O- + Pi Go' = -49.3 kJ•mol -1
O-
O O O
C-O-P-O- + ADP + Pi C-O- + ATP Go' = -18.8 kJ•mol -1
O-
Reaction 8: isomerization of 3-phosphoglycerate to 2-
phosphoglycerate
COO- phosphoglycerate COO-
mutase
H C OH H C OPO32-
CH2 OPO32- CH2 OH
3-Phosphoglycerate 2-Phosphoglycerate
Reaction 9: dehydration of 2-phosphoglycerate

COO- COO-
enolase
H C OPO32- C OPO32- + H2 O
Mg2+
CH2 OH CH2
2-Phosphoglycerate Phosphoenolpyruvate
Reaction 10: phosphate transfer to ADP
stage 1: transfer of the phosphate group

COO- O
pyruvate
kinase
C OPO32- + -O-P-O-AMP
Mg2+
CH2 O-
Phosphoenol- ADP
pyruvate COO- O O
C-OH + -O-P-O-P-O-AMP
CH2 O- O-
ATP
Enol of
pyruvate
stage 2: enolization to pyruvate

COO- COO-
C-OH C=O
CH2 CH3
Enol of pyruvate Pyruvate
Glycolysis
Summing these 10 reactions gives the net
equation for glycolysis

+ 2- glycolysis
C6 H12 O6 + 2NAD + 2HPO4 + 2ADP
Glucose
O
2CH3 CCOO- + 2NADH + 2ATP + 2H2O + 2H+
Pyruvate
Energetics of Glycolysis
Three reactions exhibit particularly large
decreases in free energy; the enzymes that
catalyze these reactions are sites of allosteric
control
hexokinase
phosphofructokinase
pyruvate kinase
Fructose is phosphorylated by fructokinase
(liver) or hexokinase (adipose) on the 1 or 6
positions resp.
Fructose-6-phosphate is an intermediate of
glycolysis.
Fructose-1-phosphate is acted upon by an aldolase-
like enz that gives DHAP and glyceraldehyde.
DHAP is a glycolysis intermediate and
glyceraldehyde can be phosphorylated to
glyceraldehyde-3-P.
Glycerol is phosphorylated to G-3-P which is
then converted to glyceraldehyde 3 phosphate.
Galactose has a slightly complicated multi-step
pathway for conversion to glucose-1-phosphate.
gal gal-1-P UDP-gal UDP-glc glc-1-P.
If this pathway is disrupted because of defect in one or
more enz involved in the conversion of gal to glc-1-P, then
galactose accumulates in the blood and the subject suffers
from galactosemia which is a genetic disorder, an inborn
error of metabolism.
Reactions of Pyruvate
Pyruvate does not accumulate in cells, but rather
undergoes one of three enzyme-catalyzed reactions,
depending of the type of cell and its state of
oxygenation
reduction to lactate
reduction to ethanol
oxidation and decarboxylation to acetyl-CoA
Lactate Fermentation
 In vertebrates under anaerobic conditions, the
most important pathway for the regeneration of
NAD+ is reduction of pyruvate to lactate
O lactate
dehydrogenase
CH3 CCOO- + NADH + H+
Pyruvate
OH
CH3 CHCOO- + NAD+
Lactate

 lactate dehydrogenase (LDH) is a tetrameric isoenzyme

consisting of H and M subunits; H4 predominates in
heart muscle, and M4 in skeletal muscle
Pyruvate to Lactate
while lactate fermentation allows glycolysis to
continue, it increases the concentration of lactate
and also of H+ in muscle tissue

lactate OH
C6 H12 O6 fermentation 2CH3 CHCOO- + 2H+
Glucose Lactate

when blood lactate reaches about 0.4 mg/100 mL,

muscle tissue becomes almost completely
exhausted
 Pyruvate to Ethanol
Yeasts and several other organisms regenerate NAD +
by this two-step pathway
decarboxylation of pyruvate to acetaldehyde

pyruvate O
O decarboxylase
- +
CH3 CCOO + H CH3 CH + CO2
Pyruvate Acetaldehyde

reduction of acetaldehyde to ethanol

alcohol
O
dehydrogenase
CH3 CH + NADH + H+
Acetaldehyde CH3 CH2 OH + NAD+
Ethanol
Pyruvate to Acetyl-CoA
Under aerobic conditions, pyruvate undergoes
oxidative decarboxylation
the carboxylate group is converted to CO2
the remaining two carbons are converted to the acetyl
group of acetyl-CoA

O oxidative
decarboxylation
CH3 CCOO- + NAD+ + CoASH
Pyruvate
O
CH3 CSCoA + CO2 + NADH
Acetyl-CoA
Read up Chapter 17