CHAPTER 7 :

GLYCOLYSIS, TCA CYCLE AND

OXIDATIVE PHOSPHORYLATION

Learning outcomes
Recognize the precursor and products in glycolsis.
Relate glycolysis to TCA cycle.
Relate NADH and FADH2 to carbohydrate

metabolism.
Relate TCA cycle to anabolism and catabolism.
Relate TCA cycle to electron transport chain.
Understand oxidative phosphorylation in
mitochondria.

Glycolysis
Glycolysis: a series of 10 enzyme-catalyzed

reactions by which glucose is oxidized to two

molecules of pyruvate
O
C6 H12 O6
Glucose

glycolysis

2CH3 CCOO- + 2H+

Pyruvate

there is net conversion of 2ADP to 2ATP

C6 H12 O6 + 2ADP + 2Pi

Glucose

O
2CH3 CCOO- + 2ATP
Pyruvate

Fates of
pyruvate

Summing these 10 reactions gives the net

equation for glycolysis

C6 H12 O6 + 2NAD+ + 2HPO4 2- + 2ADP

Glucose

glycolysis

O
2CH3 CCOO- + 2NADH + 2ATP + 2H2O + 2H+
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
O
CH3 CCOOPyruvate

anaerobic conditions
contracting muscle
anaerobic conditions
fermentation in yeast

3CO2 + 2H2 O
OH
CH3 CHCOOLactate
CH3 CH2 OH + CO2
Ethanol

Glycolysis - Reaction 1
Reaction 1: phosphorylation of -D-glucose
HO
HO

CH2 OH
O

OH
-D-Glucose OH

O
O
+ -O- P-O- P-O-AMP

HO
HO

O- OATP
CH2 OPO3 2O

hexokinase
Mg 2+

O
+ -O- P-O-AMP
O-

OH
OH
-D-Glucose-6-phosphate

ADP

Glycolysis - Reaction 2
Reaction 2: isomerization of glucose-6-

phosphate to fructose-6-phosphate
6

HO
HO

CH2 OPO3
O

2-

phosphoglucoisomerase

OH
OH
-D-Glucose-6-phosphate

CH2 OPO3 2- 1
CH2 OH
O
H HO
2
H
OH
HO
H
-D-Fructose-6-phosphate

this isomerization is most easily seen by

considering the open-chain forms of each

monosaccharide; it is one keto-enol tautomerism
followed by another
1

H
HO
H
H

CHO
2
OH
H
OH
OH
CH2 OPO32-

Glucose-6-phosphate

1 CH OH
2
2C O

H C OH
C OH
HO
H
H
OH
H
OH
CH2 OPO32(An enediol)

HO
H
H

H
OH
OH
CH2 OPO32Fructose-6-phosphate

Glycolysis - Reaction 3
Reaction 3: phosphorylation of fructose-6-

phosphate
6

CH2 OPO3 2- 1
CH2 OH
O
H HO
+ ATP
H
OH
HO
H
-D-Fructose-6-phosphate

phosphofructokinase
Mg 2+
6

CH2 OPO3 2- 1
CH2 OPO3 2O
+ ADP
H HO
H
OH
HO
H
-D-Fructose-1,6-bisphosphate

Glycolysis - Reaction 4
Reaction 4: cleavage of fructose-1,6-

bisphosphate to two triose phosphates

CH2 OPO3

C=O

C=O
HO
H
H

aldolase

H
OH
OH
CH2 OPO3

CH2 OPO3 2-

2-

CH2 OH
CHO

2-

Fructose-1,6-bisphosphate

H C OH
CH2 OPO3 2-

Dihydroxyacetone
phosphate

D-Glyceraldehyde
3-phosphate

Glycolysis - Reaction 5
Reaction 5: isomerization of triose phosphates
catalyzed by triosephosphate isomerase
reaction involves two successive keto-enol
tautomerizations
only the D enantiomer of glyceraldehyde 3phosphate is formed
CH2 OH
C=O

CHOH
C-OH

CH2 OPO3 2-

CH2 OPO3 2-

CH2 OPO3 2-

An enediol
intermediate

D-Glyceraldehyde
3-phosphate

Dihydroxyacetone
phosphate

CHO
H C OH

Glycolysis - Reaction 6
Reaction 6: oxidation (and phosphorylation) of

the -CHO group of D-glyceraldehyde-3-phosphate

to give 1,3-biphosphoglycerate.
the -CHO group is oxidized to a carboxyl group
the oxidizing agent, NAD+, is reduced to NADH

A two-electron oxidation
O
G- C- H + H2 O

O
G- C- OH + 2H+ + 2e-

A two-electron reduction
NAD+ + H+ + 2eO
G- C- H + H2 O + NAD+

NADH
O
G- C- OH + H+

Glycolysis - Reaction 7
Reaction 7: transfer of a phosphate group from

1,3-bisphosphoglycerate to ADP
this reaction is called substrate-level

phosphorylation
O
C-OPO3 2-

O
+ -O-P-O-AMP
H C OH
OCH2 OPO321,3-BisphosphoADP
glycerate

phosphoglycerate kinase

Mg2+

COOH C OH
CH2 OPO32-

3-Phosphoglycerate

O O
+ -O-P-O-P-O-AMP
O- OATP

Glycolysis Reaction 8
Isomerization of 3-phosphoglycerate to give

2-phosphoglycerate.
1
2
3

Glycolysis - Reaction 9
Reaction 9: dehydration of 2-

phosphoglycerate
COOH C

OPO32-

CH2 OH
2-Phosphoglycerate

enolase
Mg2+

COOC OPO32- + H2 O
CH2

Phosphoenolpyruvate

Glycolysis - Reaction 10
Reaction 10: phosphate transfer to ADP to give

pyruvate.
COOC OPO32CH2
Phosphoenolpyruvate

O
+ -O-P-O-AMP
O-

pyruvate
kinase
Mg2+

ADP
COO-

O
O
C-OH + -O-P-O-P-O-AMP
O- OCH2
ATP
Enol of
pyruvate

Reactions of Pyruvate
Pyruvate does not accumulate in cells, but

rather undergoes one of three enzymecatalyzed reactions, depending of the type of

cell and its state of oxygenation
reduction to lactate
reduction to ethanol
oxidation and decarboxylation to acetyl-CoA

Pyruvate to Lactate
In vertebrates under anaerobic conditions, the most

important pathway for the regeneration of NAD+ is

reduction of pyruvate to lactate
O
CH3 CCOO- + NADH + H+

Pyruvate

lactate
dehydrogenase
OH
CH3 CHCOO- + NAD+

Lactate

C6 H12 O6
Glucose

lactate
fermentation

OH
2CH3 CHCOO- + 2H+
Lactate

Pyruvate to Ethanol
Yeasts and several other organisms regenerate

NAD+ by this two-step pathway

decarboxylation of pyruvate to acetaldehyde
pyruvate
O
decarboxylase
+
CH3 CCOO + H
Pyruvate

O
CH3 CH + CO2

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

oxidative
O
decarboxylation
CH3 CCOO- + NAD+ + CoASH
Pyruvate

O
CH3 CSCoA + CO2 + NADH
Acetyl-CoA

Formation of Acetyl-CoA
a

The Citric Acid Cycle

Pyruvate

NAD+
NADH
Coenzyme A
Acetyl -CoA
NADH
NAD+

Citric
acid
cycle
(8 steps)

FADH2

NADH
CO2
NAD+

FAD
GTP

NAD+

GDP

NADH
CO2

The Citric Acid Cycle

Step 1: condensation of acetyl-CoA with

oxaloacetate;
O
CH3 C-SCoA
citrate
Acetyl-CoA
synthase
+
O C-COO
CH2 -COOOxaloacetate

CH2 -COO-

HO C-COO- + CoA-SH
CH2 -COO- Coenzyme A
Citrate

The Citric Acid Cycle

Step 2: dehydration and rehydration gives

isocitrate; catalyzed by aconitase

CH2 -COOHO C-COOCH2 -COOCitrate

CH2 -COOC-COOCH-COOAconitate

CH2 -COOH C-COOHO CH-COOIsocitrate

only one of the 4 stereoisomers of isocitrate is

formed in the cycle

The Citric Acid Cycle

Step 3: oxidation of isocitrate followed by

decarboxylation

CH2 -COO- NAD+

H C-COO-

HO CH- COO
Isocitrate

NADH

isocitrate
dehydrogenase
CH2 -COOH C-COOO C-COOOxalosuccinate

CO2

CH2 -COOH C-H

O C-COO-Ketoglutarate

isocitrate dehydrogenase is an allosteric enzyme; it is

inhibited by ATP and NADH, activated by ADP and NAD +

The Citric Acid Cycle

Step 4: oxidative decarboxylation of -

ketoglutarate to succinyl-CoA
CoA-SH
CH2 -COOCH2
O C-COO-Ketoglutarate

NAD+

NADH

-ketoglutarate
dehydrogenase
complex

CH2 -COOCH2

+ CO2

O C SCoA
Succinyl-CoA

like pyruvate dehydrogenase, this enzyme is a

multienzyme complex and requires coenzyme A,

thiamine pyrophosphate, lipoic acid, FAD, and NAD+

The Citric Acid Cycle

Step 5: formation of succinate
CH2 -COOCH2
O C SCoA

Succinyl-CoA

+ GDP + Pi

succinyl-CoA
synthetase CH2 -COO-

CH2 -COO

+ GTP + CoA-SH

Succinate

this is the first energy-yielding step of the cycle

the overall reaction is slightly exergonic

The Citric Acid Cycle

Step 6: oxidation of succinate to fumarate
CH2 -COO-

FAD

CH2 -COO-

succinate
dehydrogenase

Succinate

FADH2
-

OOC

C
C

COOH

Fumarate

Step 7: hydration of fumarate

H
-

C
C

COO-

OOC
H
Fumarate

H2 O
fumarase

HO CH-COOCH2 -COOL-Malate

The Citric Acid Cycle

Step 8: oxidation of malate
HO CH-COOCH2 -COOL-Malate

NAD+ NADH
malate
dehydrogenase

O C-COO

CH2 -COOOxaloacetate

From Pyruvate to CO2

Pyruvate dehydrogenase complex
Pyruvate + CoA-SH + NAD +
Acetyl-CoA + NADH + CO2 + H+
Citric acid cycle
Acetyl-CoA + 3NAD + + FAD + GDP + Pi + 2H2 O
2CO2 + CoA-SH + 3NADH + 3H+ + FADH2 + GTP
Pyruvate + 4NAD + + FAD + GDP + Pi + 2H2 O
3CO2 + 4NADH + FADH2 + GTP + 4H+

TCA Cycle in
The catabolism of proteins, carbohydrates,
Catabolism
and fatty acids all feed into the citric acid
cycle at one or more points
Proteins
Amino Acids

Carbohydrates

Fatty Acids

Pyruvate
Acetyl-CoA
-Ketoglutarate
Succinyl-CoA
Fumarate
Malate
Oxaloacetate

intermediates
of the citric
acid cycle

TCA Cycle in Anabolism

The citric acid cycle is the source of starting

materials for the biosynthesis of other

compounds (protein)
examples:

O
-

OOCCH2 CH2 CCOO

transamination

NH3 +
-

OOCCH2 CH2 CHCOOGlutamate

-Ketoglutarate
O
transamination
OOCCH2 CCOO
Oxaloacetate

NH3 +
-

OOCCH2 CHCOOAspartate

Electron Transport Chain

Carried out by four (4) closely related multisubunit

membrane-bound complexes and two

carriers, coenzyme Q and cytochrome c

electron

electrons from FADH2 and NADH are transferred from

one complex to the next until they reach O2

O2 is reduced to H2O

O2 + 4H+ + 4e-

2H2 O

as a result of electron transport, protons are pumped

across the inner membrane to the intermembrane space

the proton gradient establishes a voltage gradient
the proton and voltage gradients together provide

the mechanism to couple electron transport with

phosphorylation of ADP

Proton gradient across inner mitochondrial membrane

From TCA cycle

From glycolysis
and TCA cycle

Complex I
NADH-CoQ oxidoreductase
electrons are passed from NADH to FMN
H3 C

H3 C

2H+ + 2e-

O
NH
N

CH2
(CHOH) 3
CH2 OPO32 Flavin mononucleotide
(FMN)

H3 C
H3 C

H
N

O
NH

O
N
N
CH2 H
(CHOH) 3
CH2 OPO32 -

Dihydroflavin mononucleotide
(FMNH2 )

Complex I
electrons are then passed to the iron-sulfur

clusters
finally, electrons are passed to coenzyme Q
(also
called ubiquinone)
O
CH3 O

CH3

CH3 O

(CH2 CH=CCH2 ) n H + 2H+ + 2e-

CH3

O
Coenzyme Q
(oxidized form)

reduction
oxidation

OH
CH3 O

CH3

CH3 O

(CH2 CH=CCH2 ) n H

CH3

OH
Coenzyme QH2
(reduced form)

Complex I
the overall equation for the reaction of complex I

is

NADH + H+ + E-FMN
E-FMNH2 + 2Fe-S( ox)
Fe-S( red) + CoQ + 2H+
NADH + H+ + CoQ

NAD+ + E-FMNH2
E-FMN + 2Fe-S( red) + 2H+
Fe-S( ox) + CoQH2
NAD+ + CoQH2

G' = -81 kJmol-1

this transfer of electrons is strongly exergonic

and is sufficient to drive the phosphorylation of

-1
ADP
+
ADP Pi
ATP + H2 O G' = +30.5 kJmol

Complex II
Succinate-CoQ oxidoreductase
COOCH2
+ E-FAD
CH2
COOSuccinate
E-FADH2 + CoQ
Succinate + CoQ

H
-

OOC

C
C

COO+ E-FADH2
H

Fumarate
E-FAD + CoQH2
Fumarate + CoQH2

G' = -13.5 kJmol-1

the overall reaction is exergonic, but not enough to

drive ATP production

no H+ is pumped out of the matrix during this step

Complex III
CoQH2-cytochrome c oxidoreductase

CoQH2 + 2Cyt c[Fe( III)]

CoQ + 2Cyt c[Fe(II)] + 2H+

G' = -34.2 kJmol-1

this decrease in free energy is sufficient to drive

the phosphorylation of ADP to ATP (G0 = +

30.5 kJmol-1)

Complex IV
Cytochrome oxidase:
2Cyt c[Fe( II)] + 2H+ + 1 O2
2
2Cyt c[Fe( III)] + H2 O

G' = -110 kJmol-1

complex IV contains cytochrome a, cytochrome

a3, and Cu(II), which are also involved in the

electron transport
complex IV is the link with molecular oxygen
NADH + H+ + 1 O2
NAD+ + H2 O
2

Mechanism of Ox/Phos
The mechanism by which the proton gradient

leads to the production of ATP depends on ion

channels through the inner mitochondrial
membrane
protons flow back into the matrix through
channels
the flow of protons is accompanied by
formation of ATP

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