Biological

Oxidation

Biological oxidation is the

cellular process in which the
organic substances release energy
(ATP), produce CO2 and H2O
through oxidative-reductive
reactions.
organic substances:
carbohydrate, fat and protein

Principal of Redox Reaction

The electron-donating molecule in a
oxidation-reduction reaction is called
the reducing agent or reductant;
the electron-accepting molecule is the
oxidizing agent or oxidant:
for example:
Fe2+ (ferrous) lose -e
Fe3+ (ferric) gain +e

Redox reaction = reduction-oxidation

reaction
Several forms of Biological Reduction
1. Gain of electrons
2. Hydrogenation
3. Deoxygenation

Several forms of Biological Oxidation

1. Loss of electrons
2. Dehydrogenation
3. Oxygenation

oxidation-reduction potential
( or redox potential), E : it is a
measure of the affinity of a
substance for electrons. It decide
the loss (or the gain) of electrons.
A positive E: the substance has a
higher affinity for electrons , accept
electrons easily.
A negative E: the substance has a
lower affinity for electrons , donate
electrons easily.

 E0`, the standard redox

potential for a substance :is
measured under stander
condition(25, 1mmol/L reaction
substance),at pH7, and is expressed
in volts.

Respiration Chain and

Oxidative Phosphorylation

Respiratory Chain
Term:
A chain in the mitochondria consi
sts of a number of redox carriers fo
r transferring electrons from the subs
trate to molecular oxygen to form ox
ygen ion, which combines with proto
ns to form water.

Redox carriers including 4 protein

complexes
1.Complex I:
NADH:ubiquinone oxidoreductase
NADH:CoQ oxidoreductase

2.Complex II:
Succinate dehydrogenase

3.Complex III:
cytochrome bc1 (ubiquinone Cyt c oxidoredu
ctase)

4.Complex IV:
cytochrome oxidase

Complex I (NADH:ubiquinone
oxidoreductase)

Function: transfer electrons from NADH

to CoQ
Components:
NADH dehydrogenase (FMN)
Iron-sulfur proteins (Fe-S)
complex
NADH
CoQ

FMN; Fe-SN-1a,b; Fe-SN-4; Fe-SN-3; Fe-SN-2

R=H: NAD+;

R=H2PO3:NADP+

1. NAD(P)+: Nicotinamide Adenine Dinucleotide Phosphate)

Oxidation of NADH is a 2-electron

(2e), 2-proton(2H) reaction

NAD+ or NADP+

NADH or NADPH

2. FMN can transfer 1 or 2 hydride i

ons each time
FMN: flavin mononucleotide

Accepts 1 H+ and 1 eto form semiquinone

= stable free radical

Accepts 2 H+ and 2 eto give fully reduced form

3. Iron-sulfur clusters (Fe-S) transf

ers 1-electron at a time, without pro
ton involved
Fe3++e-

Fe2+

4.Ubiquinone (CoQ) is lipid-soluble, not

a component of complex , can transfer 1 o

r 2 hydride ions each time.
Function: transfer electrons and protons
from complex , to complex .

NADH+H+
NAD+

FMN

Reduced Fe-S

FMNH2

Oxidized Fe-S

QH2

Matrix

Intermembrane space

Complex II: Succinate dehydrogen

ase (Succinate: CoQ oxidoreductas
e)

Function: transfer electrons from suc

cinate to CoQ
Components:
Succinate dehydrogenase (FAD, FeS)
Cytochrome b560
Complex
Succinate Fe-S1; b560; FAD; Fe-S2 ; Fe-S3

CoQ

Cytochromes a, b, c are heme proteins,

their heme irons participate redox
reactions of e- transport.

Fe3++e-

Fe2+

Intermembrane space

Matrix

Succinate

Complex III:

cytochrome bc1 (ubiquinone Cyt c oxidor

eductase)

Function: transfer electrons from CoQ

to cytochrome c
Components: iron-sulfur protein
cytochrome b(b562, b566)
cytochrome c1
complex
QH2

b562; b566; Fe-S; c1

Cyt c

Cytochrome c is soluble, which will transfer

electrons to complex
Intermembrane
space

Matrix

Complex IV: cytochrome oxida

se

Function: transfer electrons from Cyt c

to molecule oxygen, the final electron
acceptor.
Components: cytochrome aa3
copper ion (Cu2+)
Cu2+ + e-

Cu+

Complex IV
Cyt c CuAaa3CuB

O2

Cytochrome c
Coenzyme Q
ubiquinone/ol

Sequence of respiratory chai

n
Principles:
e- tend to flow from a redox pair with a lowe
r Eto one with a higher E
In the e--transport chain, e--carriers are arra
nged in order of increasing redox potential,
making possible the gradual release of ener
gy stored in NADH, FADH2

Redox potential

redox pair
NAD+/NADH+H+
FMN/ FMNH2
FAD/ FADH2
Cyt b Fe3+/Fe2+
Q10/Q10H2
Cyt c1 Fe3+/ Fe2+
Cyt c Fe3+/Fe2+
Cyt a Fe3+ / Fe2+
Cyt a3 Fe3+ / Fe2+
1/2 O2/ H2O

E'
E 0(V)
-0.32
-0.30
-0.06
0.040.10
0.07
0.22
0.25
0.29
0.55
0.82

There are two respiratory chains

NADH respiratory chain
NADH
Complex
CoQ
Comple
x
cytochrome c
Complex
O2
Succinate (FADH2) respiratory chain
Succinate Complex CoQ Complex
cytochrome c Complex O2

NADH
respiration
chain

FADH2
respiration
chain

Oxidative Phosphorylation
The oxidation of organic nutritions produces the
energy-rich molecules, NADH and FADH2.
The oxidation of NADH or FADH2 in mitochondri
al is the electron transferring through respiratio
n chain.
The free energy produced in electron transferrin
g supports the phosphorylation of ADP to form
ATP.
The oxidation of NADH or FADH2 and the format
ion of ATP are coupled process, called Oxidation
Phosphorylation.

The Chemiosmotic Theory

The free energy of electron transport i
s conserved by pumping protons from
the mitochondrial matrix to the interm
embrane space so as to create an elec
trochemical H+ gradient across the inn
er mitochondrial membrane. The elec
trochemical potential of this gradient i
s harnessed to synthesize ATP.
Peter Mitchell

Electrochemical H+ gradient (Proto

n-motive force)
2 components involve
d
1. Chemical potential energy
due to difference in [H+]
in two regions separated
by a membrane
2. Electrical potential energy
that results from the separ
ation of charge when a pr
oton moves across the me
mbrane without a electro
n.

Complex I:
4 H+ expelled
per e--pair
transferred to
Q

Complex III:
4 H+ expelled per
e--pair transferred
to Cyt c

Complex IV:
2e- + 2 H+ from
matrix convert O2
to H2O; 2 further H+
expelled from

Proton pumping: Reductiondependent conformational switch of an

e--transport complex

Conformation 1
(high affinity for H+)

Conformation 2
(low affinity for H+).

ATP Synthase
Intermembrane space

Inner
(ab2c9-12)

Membrane

Matrix

C ring

(33
)

-subunit take up ADP and Pi to form ATP

ADP + Pi

ATP
Each of 3 -subunits
contains an active
site

F1: multisubunit
complex that catalyzes
ATP synthesis

F0 = proton-conducting
transmembrane unit

When protons flow back through F0 channel, -subunit is

rotated by the rotation of c ring, then the conformations of
-subunits are changed, this lead to the synthesis and
release of ATP. To form a ATP need 3 protons flow into
matrix.

H+ flow

-subunit has three conformations:T (tight), L (loose), O (open)

Translocation of ATP , ADP and Pi.

ADP3- ATP4-

H+

H2PO4- H+

Intermembrane

space

F0

Matrix
F1

ATP4ADP3H+

H2PO4- H+

P/O ratios
P/O ratio is the rate of phosphate inc
orporated into ATP to atoms of O2 util
ized. It measure the number of ATP
molecules formed per two electrons t
ransfer through the respiratory chai
n.
NADH respiratory chain : 2.5,
FADH2 respiratory chain: 1.5

 During two electrons transfer through

NADH respiratory chain, ten protons ar
e pumped out of the matrix.
To synthesis and translocation an ATP,
four protons are needed.
So, two electrons transport can result i
n 2.5 ATP.
To succinate respiratory chain , two ele
ctrons transport can result in 1.5 ATP.

Regulation of Oxidative Phosphoryl

ation
1.PMF (proton motive force) regulate t
he electron transport.
higher PMF
lower rate of tr
ansport
2.ADP concentration
resting condition: energy demande
d is low, ADP concentration is low, the
speed of Oxidative Phosphorytion is lo
w.
active condition: the speed is high.

Inhibitor of Oxidative Phosp

horylation
1.Inhibitor of electron transport
Succinate

Antimycin A

Cyanide, Azide
Carbon Monoxide

Retonone
Amytal

 2.Uncoupling agents
uncoupling protein (in brown adipose tissue),
2,4-dinitrophenol, Pentachlorophenol

heat

H+
Intermenbran space

Cyt c

uncoupling
protein

F0

F1

Matrix

+ H+

H+

ADP+Pi ATP

2,4-dinitrophnol

3.Oligomycin bonds at the connection of F 0

and F1, inhibit the function of ATP synthase.
Intermembrane space

Matrix

Oligomycin
C ring

Succinate

Retonone
Amytal

Antimycin A

Oligomycin

Uncoupling
agent

ATP and other Energy-rich compoun

ts
ATP has two energy-rich phosphoric acid anhydride
bonds, the hydrolysis of each bond release more
energy than simple phosphate esters.
NH 2
N

N
OH
O= P
OH

OH

OH

O p O p
OH

OCH2 O

OH

H
H

H
H

OH OH

AMP
ADP
ATP

Some Energy-rich compounds

Structure

Exemple

creatine phosphate

phosphoenolpyruvate

acetyl phosphate

Acetyl CoA

 The hydrolysis of energy-rich bond:

G = -5 -15kcal/mol
The compounds with energy-rich bond
are high-energy compounds.
The hydrolysis of low-energy bond:
G = -1 -3kcal/mol
The compounds with low energy bond
are low-energy compounds.

Transport of high-energy bond ene

rgies
1.Substrate level phosphorylation
Glycerate 1,3-biphosphate + ADP
Glycerate 3-phosphate +ATP

G = -4.5kcal/mol
Phosphoenolpyruvate +ADP
Pyruvate + ATP

G = -7.5kcal/mol

2.ATP is the center of energy producing and

utilizing.
ATP

Oxidative
Phosphorylation

Energy
utilization

Substrate level
phosphorylation

~P
~P

ADP

3.Other nucleoside triphosphates ar

e involved in energy transport.
GTP:

gluconeogenesis
protein synthesis
UTP: glycogen
CTP: lipid synthesis

4.Transport of the terminal phosph

ate bond of ATP to the other nucle
oside

Function of nucleoside diphospha

te kinase
ATP + UDP
ATP + CDP
ATP + GDP

ADP + UTP
ADP + CTP
ADP + GTP

Function of adenylate kinase

ADP + ADP

ATP + AMP

Energy from cytosolic NAD

H
A mitochondrial NADH produce 2.5 A
TP
A cytosolic NADH must be transporte
d into mitochondrial for oxidation by
two methods.
Glycerol phosphate shuttle
1.5 ATP
Malate aspartate shuttle
2.5 ATP

Glycerol phosphate shuttle

NADH+H

Glycerol
phosphate
dehydrogenase

NAD+

CH2OH

CH2OH

C=O

C=O

CH2O- Pi

CH2O- Pi

dihydroxyacetone
phosphate

dihydroxyacetone
phosphate

Electron chain

CH2OH

CH2OH

CHOH

CHOH

CH2O- Pi

CH2O- Pi

Glycerol
phosphate

Glycerol
phosphate
Intermembran
space

FADH2

FAD
Glycerol
phosphate
dehydrogenase

Inner menbran

Malate aspartate shuttle

O

Aspartate
-OOC-CH2-C-COO-

H3N
Malate
-ketoglutarate
carrier

OOC-CH2-C-COO

Aspartate

oxaloacetate

oxaloacetate
Glutamate

NADH
+H+

Glutamate

Electron chain
NADH
+H+

NAD+

-ketoglutarate

-ketoglutarate

OH

NAD+

-OOC-CH 2-C-COOH

Malate

cytosol

Glutamate-aspartate
carrier

inner mitochondrial
membran

Malate
matrix

Other Biological Oxidations

Monoxygenases
dioxygenase --add 2 atoms of O2
oxygenase to organic compounds.
monoxygenase (mixed-function oxidase,
hydroxylase)
--adds 1 oxygen atom to organic comp
ounds as a hydroxyl group.
RH + NADPH + H+ + O2

ROH + NADP+ + H2O

The chief compounds of monoxygenase:

Cyt b5, Cyt P450, Cyt P450 reductase(FAD,FMN)

Free Radical Scavenging Enzy

mes
Free Radical: the groups with an unpa
ired electron. (such as O2 H2O
2 OH)
1.Superoxide dismutases(SODs)
2O2+ 2H+

SOD

H2O2 + O2

peroxidase

H2O + O2

 2.Glutathione peroxidase
H2O2
(ROOH)

2G SH

Glutathio
ne peroxid

Glutathione
reductase

ase
H2O
(ROH+H2O)

NADP+

G S S G

NADPH+H+

 3.Catalase (in peroxisomes)

2H2O2

catalase

2H2O + O2

summary