BIOLOGICAL OXIDATION

&
PRINCIPLE OF ENERGY METABOLISM

EDITED & RECOMPOSED BY
Dr. Liniyanti D.Oswari MSc.
For Medical student, Sriwijaya University
Block 8

Citric Acid Cycle

Biological oxidation
Carbohydrate metabolism
Glucose, rbc metabolism, glycogen, blood
glucose, diabetes
Lipid metabolism
Plasma lipoproteins CM, VLDL, LDL, HDL
Protein metabolism
Gluconeogenesis

Calories
Fat contains 9 calories per gram
Protein contains 4 calories/gram
Carbohydrates has 4 calories per
gram
(approximately)
Anabolism: Building Up
ATP produced during catabolism
drives anabolism.
Excess carbohydrates energy can
result in fat synthesis.
Humans synthesize 11 of 20 amino acids;
remaining 8 essential amino acids must be
provided by diet.

Anabolism
Large complex molecules are synthesized
from smaller precursors.
Building block molecules (amino acids,
sugars and fatty acids) are produced or
acquired from the diet.
Because anabolic processes include the
synthesis of polysaccharides and proteins
from sugars and amino acids, the
biosynthetic pathways increase order and
complexity, they require inputs of free
energy (ATP and NADPH).

Metabolism in Muscle
Eric Niederhoffer
SIU-SOM
Acetyl-CoA Lactate
No O
2

Fatty acids
|-Oxidation
Production of ATP
G6P
Glucose
Glycolysis
Pyruvate
BCAA
Ile, Val
Krebs
cycle
Electron
Transport
Chain
Glycogen
Glycogenolysis
Ca
2+

PKa
Ca
2+

PDH
Ca
2+

ISDH, oKGDH
Carbohydrate metabolism
Glucose
Rbc metabolism
Glycogen
Blood glucose
Diabetes

Glucose
How does the body metabolise glucose?
How can we obtain energy from glucose?
How is energy derived from glucose?

Glucose
2 types of glycolysis:
Aerobic g. and anaerobic g.
Aerobic g. occurs when oxygen supply is sufficient
Anaerobic g. occurs when oxygen supply is lacking

In aerobic g.:
Oxygen status: sufficient oxygen supply
Glucose pyruvate TCA GTP, NADH & FADH2
Substrate-level phosphorylation: GTP ATP
NADH & FADH2 ETC ATP:
1 NADH 3 ATP
1 FADH2 2 ATP
Anaerobic g.:
Oxygen status: insufficient oxygen supply
Glucose pyruvate lactate
Lactate is used via Cori cycle
Rbc metabolism
What is the source of energy for rbc?
Rbc has no mitochondria
Rbc depends entirely on glycolysis for ATP

Glycogen
How is glycogen metabolised by the body?
How can we obtain energy from glycogen?
How is energy derived from glycogen?



Glycogen
Glycogen is involved in 2 ways:
Glycogen synthesis (glycogenesis)
Glycogen breakdown (glycogenolysis)

At high blood glucose level:
Glycogen is synthesized and stored in liver and muscles

At low blood glucose level:
Glycogen is broken down (degraded) by enzymes to yield glucose
Two enzymes breakdown glycogen to glucose:
Branching enzyme
Straight chain enzyme

Liver vs. muscle glycogen:
Body has more liver glycogen than muscle glycogen
Liver glycogen is used to maintain blood glucose level
Muscle glycogen is used internally
Blood glucose
What is normal blood glucose level?

Note:
Blood glucose is determined under fasting condition
Plasma is used to determine glucose content
Quote values in mmol/L, or mg/dl

Normal fasting plasma glucose (FPG) is 4.2-6.2
mmol/L=70 110 mg/dl
Maintenance of blood glucose
Note:
There are many factors which regulate blood glucose level
Factors: insulin, liver, glucagon, epinephrine, etc

When we eat:
At high blood glucose level, insulin is secreted
Insulin causes cells to take up glucose
Cells use glucose for energy

When we sleep:
The liver maintains blood glucose (by hepatic glycogenolysis) to
within acceptable levels between 4.2-6.2 mmol/L =70-110 mg/dl
(fasting values)
Gluconeogenesis
What is gluconeogenesis?
Formation of glucose from non-glucose sources
such as C-skeletons of glucogenic amino acids

Under what conditions does this occur?
Gluconeogenesis occurs when blood glucose is
low
Lipogenesis and Lipolysis
Figure 24.14
Protein Metabolism
Deaminated amino acids are converted into:
Pyruvic acid
One of the keto acid intermediates of the Krebs
cycle
These events occur as transamination,
oxidative deamination, and keto acid
modification
OVERVIEW OF METABOLIC PATHWAYS
Amino
Acids
Glucose-6-P
Pyruvate
ATP
Acetyl-CoA
Urea
AND SYSTEMS OF ENERGY METABOLISM
b
a
o
e
h
p
Nucleic
Acids
GLYCOGEN
PROTEIN
TRIACYLGLYCEROLS
d
c
Ribose-5-P
f
g
i
j
k
l
a
b
Glucose

Lactate

n
m
Ketone
Bodies

Figure . Energy systems
Free Fatty Acids

Energy
The meaning of energy in energy
metabolism
In a haste to learn the individual reactions in a pathway, its
easy to lose sight of the purpose of the pathway. With energy
metabolism, the purpose is to generate energy, generally as ATP or
NADH or some high energy compound that will be used in a later
anabolic step. Glycolysis and Krebs cycle reactions have a high
number of kinase and dehydrogenase enzymes, respectively, for this
reason. This class of enzymes is intimately connected with energy
production and conservation. Pathways in the cytosol tend to be less
energy yielding, whereas those in the mitochondria are almost totally
devoted to energy production. This tutorial will bring you closer to
understanding why and how cells conserve energy. It will also help
you see the logic behind molecular energy calculations. As you listen
to your heart pump or move your arm to scratch your head, you
should be able to tell what purpose energy serves to life.
Thermodynamic Laws
The First Law of Thermodynamics.
A systems internal energy can change only by the
exchange of heat or work with the surroundings.
A Statement of Conservation of energy.

The Second Law of Thermodynamics.
The entropy of an isolated system will tend to
increase to a maximum value.
The entropy of such a system will not decrease,-
sucrose will never de-diffuse into corner of the
solution.
Entropy is a measure of the randomness or
disorder in a system.



The terms energy conservation and energy generation tend to carry the
same meaning. Conservation implies avoiding heat, or channeling the energy
differential between reactants and products into the synthesis of a compound.
Because energy as heat cannot be exploited in an isothermal system, biological
systems have to conserve energy by biosynthesis. Suppose for example ATP is
hydrolyzed during a reaction (click 1). The standard energy differential between
reactants and products (AG
o
) of that reaction is 30.5 kJ/mol.
What is energy conservation?
This means the environment of the cell gains 30.5 kJ of heat energy for each
mole of ATP hydrolyzed by water. Obviously, this is wasteful. To counter the
loss, ATP hydrolysis is coupled with the synthesis of a phosphorylated
compound. You saw this as a coupled reaction when ATP was needed to
produce glucose-6-PO
4
or fructose 1,6-bisPO
4
(click 1).
Now you see that by making glucose-6-PO
4
or fructose 1,6-bisPO
4
, the cell avoids
losing the larger part of the ATP hydrolysis energy as heat. This is energy
conservation. Click one to go on.
ATP + H
2
O ADP + PO
4
Glucose + ATP Glucose-6-PO
4
+ ADP
Fructose-6-PO
4
+ ATP Fructose 1,6-bisPO
4
+ ADP
LIFE opposes ENTROPY, S:
a) Entropy & energy: heat exchange--
100
o
C
25
o
C
25
o
C
25
o
C
37
o
C
25
o
C
x
25
o
C
25
o
C
2nd Law of Thermodynamics:
Direct vs Indirect Energy Production
The energy generated in metabolic pathways comes in two forms,
direct or indirect. Direct or substrate level refers to energy generated during
a particular reaction. The production of ATP by reacting ADP with PEP is an
example of this type (click 1)
COO
CH
3
C=O
COO
CH
2
C~OPO
3
=
+ ADP
+ ATP
Indirect refers to energy channeled into a compound that will return
the energy at a later step. High energy compounds such as acyl-phosphates
or thioesters fit this example. Another is NADH generated during oxidation
reactions in the cytosol or Krebs cycle. When L-malate is oxidzed by NAD
+
,
NADH is generated (click 1). NADH and FADH
2
have trapped the electron
pair from the oxidation in their structures and will release the energy when
they themselves are oxidized.
COO
CH2
HO-C-H
COO
C=O
COO
CH2
COO
+ NAD
+
+ NADH + H
+
:
:
Calculating energy yield in glycolysis
Calculating energy yield helps you see the energy phase of metabolism
in real numbers. Take for example the energy yield when glyceraldehyde-3-PO
4

is oxidized to pyruvate. How much energy is conserved in this reaction? To
determine that number we need to know the pathway. We also need to know if
anaerobic or aerobic conditions prevail. First the pathway. There are 5 enzyme-
catalyzed reactions to consider (click 1).
glyceraldehyde-3-PO
4
+ PO
4
+ NAD
+
1,3-bisPO
4
glycerate

1,3-bisPO4 glycerate
3-phosphglycerate
2-phosphoglycerate
PEP
2-phosphoglycerate
PEP
pyruvate + ATP + H
2
O
glyceraldehyde-3-PO
4
+ PO
4
+ NAD
+
+ 2ADP pyruvate + NADH + H
+
+ 2ATP + 2H
2
O
Removing the common terms on both sides yields a final equation (click 1).
We see that the phosphate on glyceraldehyde-3-PO
4
and the inorganic PO
4

both contribute to formation of ATP. Thus, 2 ATPs are formed by the 5
reactions. Under anaerobic conditions two represents the final yield. But, if
the reaction was carried out with oxygen and involved the mitochondria,
energy to the equivalent of 5 ATPs would result. Click 1 to see why.
+ NADH + H
+
3-phosphoglycerate
+ ATP
+ ADP
+ ADP
+ H
2
O
Energy yield in the mitochondria
The mitochondria is the heart of aerobic metabolism. Electrons in
NADH and FADH
2
are channeled into the electron transport system, which is
driven by O
2
. A large part of energy of oxidation of the electron transport
components is preserved in ATP. Each NADH generates the equivalent of 3
ATPs and each FADH
2
, 2 ATPs for each pair of electrons transferred to oxygen
(click 1).
O
2
H
2
O
NAD
NADH Electron transport
ATP ATP ATP
NADH from the cytosol yields its electrons indirectly via a shuttle. NADH
generated by the 3 NAD-linked dehydrogenases in the Krebs cycle provide
most of the energy. For example, each citrate molecule oxidized to CO
2
and
H
2
O generates the equivalent of 36 ATPs. Click 1 to see how this value was
obtained.
:
Energy yield in the Krebs cycle
A cycle implies the last intermediate returns to the front. Each turn of the
Krebs cycle results in the loss of 2 carbons as CO
2
and generates 3NADH, one
FADH
2
and one GTP (click 1). A 2-carbon compound, such as the acetate group
on acetyl-CoA, therefore, yields 12 ATPs of energy.
Acetyl-CoA
citrate
isocitrate
o-ketoglutarate
succinyl-CoA
succinate
fumarate
malate
oxaloacetate
NADH
NADH
GTP
FADH
2
NADH
CO
2
CO
2
Now, suppose instead of acetyl-
CoA we want to determine the ATP yield
when oxaloacetate (OAA) is oxidized (click
1). First write the equation for the oxidation
(click 1). OAA yields 4 moles of CO
2
for each
mole oxidized. Thus, 2 turns of the cycle are
needed to oxidize all of the carbons in OAA to
CO
2
. Two turns is equivalent to 24 ATPs.

Performing the same analysis for
citrate shows 6CO
2
liberated, or 3 turns of the
cycle (click 1). Thus, citrate yield 36 ATPs, or
one third more energy than OAA. Finally lets
consider the oxidation of malate (click 1).
Malate has 4 carbons, which means the
oxidation will generate 4CO
2
. But, we also
need to oxidize malate to OAA, which
generates one NADH. Thus 3 more ATPs
than OAA, i.e., 24 + 3= 27 ATPs. Click 1 to
test and expand your understanding.

C
4
H
4
O
5
+ 3
1/2
O
2
4CO
2
+ 2H
2
O
C
6
H
8
O
7
+ 4
1/2
O
2
6CO
2
+ 4H
2
O
C
4
H
6
O
5
+ 5 O
2
4CO
2
+ 3H
2
O
Free energy of a reaction
The free energy change (AG) of a reaction
determines its spontaneity. A reaction is spontaneous
if AG is negative (if the free energy of products is less
than that of reactants).
AG
o
' = standard free energy change (at pH 7, 1M
reactants & products); R = gas constant; T = temp.
For a reaction A + B C + D
AG = AG
o
' + RT ln
[C] [D]
[A] [B]
PHYSIOLOGICAL FREE ENERGY

AG = actual free energy difference of reaction in cell
AG = AG + 2.3 log [products/substrates]
"mass action effect"
change AG by altering ratio of products/substrates
product and/or reactant lowers AG
reaction proceeds better towards product
AG
o
' of a reaction may be positive, & AG negative,
depending on cellular concentrations of reactants and
products.
Many reactions for which AG
o
' is positive are
spontaneous because other reactions cause depletion
of products or maintenance of high substrate
concentration.
For a reaction A + B C + D
AG = AG' + RT ln
[A] [B]
[C] [D]
Equilibrium versus Steady State
Steady state
constant flux in one direction
Equilibrium
no net flux in any direction
A B C
A B


A B C
A B


A
Equilibrium constant:
fixed ratio of products:substrates when AG = 0
AG = AG + 2.3 log [products/substrates]
AG = - 2.3 log [Keq]
At equilibrium
AG = 0.
K'
eq
, the ratio
[C][D]/[A][B] at
equilibrium, is the
equilibrium constant.
An equilibrium constant
(K'
eq
) greater than one
indicates a spontaneous
reaction (negative AG').
AG = AG' + RT ln
0 = AG' + RT ln
AG' = - RTln
defining K'
eq
=
AG' = - RT ln K'
eq
[C] [D]
[A] [B]
[C] [D]
[A] [B]
[C] [D]
[A] [B]
[C] [D]
[A] [B]
K'
eq
AG

'
kJ/mol
Starting with 1 M reactants &
products, the reaction:
10
4
- 23
proceeds forward (spontaneous)
10
2
- 11
proceeds forward (spontaneous)
10
0
= 1 0 is at equilibrium
10
-
2
+ 11 reverses to form reactants
10
-
4
+ 23 reverses to form reactants
AG
o
' = RT ln K'
eq
Variation of equilibrium constant with AG
o
(25
o
C)
Energy coupling
A spontaneous reaction may drive a non-spontaneous
reaction.
Free energy changes of coupled reactions are
additive.
A. Some enzyme-catalyzed reactions are interpretable as
two coupled half-reactions, one spontaneous and the
other non-spontaneous.
At the enzyme active site, the coupled reaction is
kinetically facilitated, while individual half-reactions are
prevented.
Free energy changes of half reactions may be
summed, to yield the free energy of the coupled
reaction.
For example, in the reaction catalyzed by the Glycolysis
enzyme Hexokinase, the half-reactions are:
ATP + H
2
O ADP + P
i
AG
o
' = 31 kJ/mol
P
i
+ glucose glucose-6-P + H
2
O AG
o
' = +14 kJ/mol
Coupled reaction:
ATP + glucose ADP + glucose-6-P AG
o
' = 17 kJ/mol
The structure of the enzyme active site, from which H
2
O is
excluded, prevents the individual hydrolytic reactions, while
favoring the coupled reaction.
B. Two separate reactions, occurring in the same cellular
compartment, one spontaneous and the other not, may be
coupled by a common intermediate (reactant or
product).
A hypothetical, but typical, example involving PP
i
:
Enzyme 1: A + ATP B + AMP + PP
i

AG
o
' = + 15 kJ/mol
Enzyme 2: PP
i
+ H
2
O 2 P
i

AG
o
' = 33 kJ/mol
Overall spontaneous reaction: AG
o
' = 18 kJ/mol
A + ATP + H
2
O B + AMP + 2 P
i

Pyrophosphate (PP
i
) is often the product of a
reaction that needs a driving force. Its spontaneous
hydrolysis, catalyzed by Pyrophosphatase enzyme, drives
the reaction for which PP
i
is a product.
Compounds with high energy bonds are said to have
high group transfer potential.
For example, P
i
may be spontaneously cleaved from
ATP for transfer to another compound (e.g., to a
hydroxyl group on glucose).
Potentially, 2 ~P bonds can be cleaved, as 2
phosphates are released by hydrolysis from ATP.
AMP~P~P AMP~P + P
i
(ATP ADP + P
i
)
AMP~P AMP + P
i
(ADP AMP + P
i
)
Alternatively:
AMP~P~P AMP + P~P (ATP AMP + PP
i
)
P~P 2 P
i
(PP
i
2P
i
)

High energy bonds:
ATP often serves as an energy source.
Hydrolytic cleavage of one or both of the "high energy"
bonds of ATP is coupled to an energy-requiring (non-
spontaneous) reaction.
AMP functions as an energy sensor & regulator of
metabolism.
When ATP production does not keep up with needs, a
higher portion of a cell's adenine nucleotide pool is AMP.
AMP stimulates metabolic pathways that produce ATP.
Some examples of this role involve direct allosteric
activation of pathway enzymes by AMP.
Some regulatory effects of AMP are mediated by the
enzyme AMP-Activated Protein Kinase.
A reaction important for equilibrating ~P among adenine
nucleotides within a cell is that catalyzed by Adenylate
Kinase:
ATP + AMP 2 ADP
The Adenylate Kinase reaction is also important because
the substrate for ATP synthesis, e.g., by mitochondrial ATP
Synthase, is ADP, while some cellular reactions
dephosphorylate ATP all the way to AMP.
The enzyme Nucleoside Diphosphate Kinase (NuDiKi)
equilibrates ~P among the various nucleotides that are
needed, e.g., for synthesis of DNA & RNA.
NuDiKi catalyzes reversible reactions such as:
ATP + GDP ADP + GTP,
ATP + UDP ADP + UTP, etc.
Inorganic polyphosphate
Many organisms store energy as inorganic
polyphosphate, a chain of many phosphate residues
linked by phosphoanhydride bonds:
P~P~P~P~P...
Hydrolysis of P
i
residues from polyphosphate may be
coupled to energy-dependent reactions.
Depending on the organism or cell type, inorganic
polyphosphate may have additional functions.
E.g., it may serve as a reservoir for P
i
, a chelator of
metal ions, a buffer, or a regulator.
Why do phosphoanhydride linkages have a high AG
of hydrolysis? Contributing factors for ATP & PP
i

include:
Resonance stabilization of products of
hydrolysis exceeds resonance stabilization of the
compound itself.
Electrostatic repulsion between negatively
charged phosphate oxygen atoms favors
separation of the phosphates.
Creatine Kinase catalyzes:
Phosphocreatine + ADP ATP + creatine
This is a reversible reaction, though the equilibrium constant slightly
favors phosphocreatine formation.
Phosphocreatine is produced when ATP levels are high.
When ATP is depleted during exercise in muscle, phosphate is
transferred from phosphocreatine to ADP, to replenish ATP.

O P
H
N C
O
O

N
NH
2
+
CH
2
CH
3
C
O
O

phosphocreatine
Phosphocreatine (creatine
phosphate), another
compound with a "high
energy" phosphate linkage, is
used in nerve & muscle for
storage of ~P bonds.
Phosphoenolpyruvate (PEP), involved in ATP
synthesis in Glycolysis, has a very high AG of P
i

hydrolysis.
Removal of P
i
from ester linkage in PEP is spontaneous
because the enol spontaneously converts to a ketone.
The ester linkage in PEP is an exception.
C
C
O O

OPO
3
2
CH
2
C
C
O O

O
CH
3
C
C
O O

OH
CH
2
ADP ATP
H
+
PEP enolpyruvate pyruvate
Generally phosphate esters, formed by splitting out
water between a phosphoric acid and an OH group,
have a low but negative AG of hydrolysis. Examples:
the linkage between the first phosphate and the
ribose hydroxyl of ATP.

N
N
N
N
NH
2
O
OH OH
H H
H
CH
2
H
O P O P O P -O
O
O- O-
O O
O-
adenine
ribose
ATP (adenosine triphosphate)
ester linkage

Other examples of phosphate esters with low but
negative AG of hydrolysis:
the linkage between phosphate & a hydroxyl
group in glucose-6-phosphate or glycerol-3-
phosphate.
glycerol-3-phosphate
CH
2
CH
CH
2
OH
HO
O P
O
O
O

H
O
OH
H
OH H
OH
CH
2
H
OH
H
1
6
5
4
3
2
O P
O
OH
OH
glucose-6-phosphate
the linkage between phosphate and the hydroxyl
group of an amino acid residue in a protein
(serine, threonine or tyrosine).
Regulation of proteins by phosphorylation and
dephosphorylation will be discussed later.
Protein OH + ATP Protein O P
O
O

+ ADP
P
i
H
2
O
Protein Kinase




Protein Phosphatase
ATP has special roles in energy coupling & P
i
transfer.
AG of phosphate hydrolysis from ATP is intermediate
among examples below.
ATP can thus act as a P
i
donor, & ATP can be synthesized
by P
i
transfer, e.g., from PEP.
Compound
AG
o
' of phosphate
hydrolysis, kJ/mol
Phosphoenolpyruvate (PEP)
61.9
Phosphocreatine
43.1
Pyrophosphate
33.5
ATP (to ADP)
30.5
Glucose-6-phosphate
13.8
Glycerol-3-phosphate
9.2
Kinetics vs Thermodynamics:
A high activation energy barrier usually causes
hydrolysis of a high energy bond to be very slow in
the absence of an enzyme catalyst.
This kinetic stability is essential to the role of ATP
and other compounds with ~ bonds.
If ATP would rapidly hydrolyze in the absence of a
catalyst, it could not serve its important roles in energy
metabolism and phosphate transfer.
Phosphate is removed from ATP only when the
reaction is coupled via enzyme catalysis to some other
reaction useful to the cell, such as transport of an ion,
phosphorylation of glucose, or regulation of an enzyme
by phosphorylation of a serine residue.
Pathway Eukaryote Prokaryote
Glycolysis

Cytoplasm Cytoplasm
Intermediate step

Cytoplasm Cytoplasm
Krebs cycle
Mitochondrial
matrix
Cytoplasm
ETC
Mitochondrial inner
membrane
Plasma
membrane
ATP produced from complete oxidation of 1
glucose using aerobic respiration











36 ATPs are produced in eukaryotes.
Pathway

By substrate-
level
phosphorylation
By oxidative
phosphorylation
From
NADH
From
FADH
Glycolysis

2 (2X3)=6 0
Intermediate
step
0 (2X3)=6
Krebs cycle 2 (6X3)=18 (2X2)=4
Total 4 30 4
Oxidation-Reduction
Oxidation: the loss of electrons
Reduction: the gain of electrons
Oxidation-reduction (redox) reaction: any
reaction in which electrons are transferred
from one species to another

Look for ions that change in charge
i.e. Zn (s) + Cu
2+
Zn
2+
(aq) + Cu (s)
The Zn lost electrons ..
It was oxidized
So the Cu was reduced!
Oxidation-Reduction
Remember:
LEO the lion says GER
Loss
Electrons
OXIDATION

Gain
Electrons
REDUCTION
Oxidation-Reduction
+4
+3
+2
+1
0
-1
-2
-3
-4

Oxidized

Reduced

You Try It

Fe
2+
Fe
3+
OXIDIZED!
Cu
2+
Cu (s)
Hint: Cu (s) is Cu
0
REDUCED!

Oxidation-Reduction
Example: if we put a piece of zinc metal in a
beaker containing a solution of copper(II)
sulfate
some of the zinc metal dissolves
some of the copper ions deposit on the zinc metal
the blue color of Cu
2+
ions gradually disappears
In this oxidation-reduction reaction
zinc metal loses electrons to copper ions

copper ions gain electrons from the zinc


Zn(s)
+
Zn
2 +
(aq) 2e
-
Zn is oxidized
Cu( s) + 2e
-
Cu
2 +
( aq)
Cu
2+
is reduced
Oxidation-Reduction
we summarize these oxidation-reduction
relationships in this way
electrons flow
from Zn to Cu
2+
+ Zn(s)
Cu
2 +
(aq)
+ Cu( s) Zn
2 +
( aq)
loses electrons;
is oxidized
gains electrons;
is reduced
gives electrons
to Cu
2+
; is the
reducing agent
takes electrons
from Zn; is the
oxidizing agent
Oxidation-Reduction
using these alternative definitions for the
combustion of methane
CH
4
(g)
+
O
2
(g) CO
2
(g)
+
H
2
O(g)
gains O and loses
H; is oxidized
gains H;
is reduced
is the reducing
agent
is the oxidizing
agent
electrons are
transferred from
carbon to oxygen
Oxidation-Reduction
Five important types of redox reactions
combustion: burning in air. The products of complete
combustion of carbon compounds are CO
2
and H
2
O.
respiration: the process by which living organisms use
O
2
to oxidize carbon-containing compounds to produce
CO
2
and H
2
O. The importance of these reaction is not
the CO
2
produced, but the energy released.
rusting: the oxidation of iron to a mixture of iron oxides


bleaching: the oxidation of colored compounds to
products which are colorless
batteries: in most cases, the reaction taking place in a
battery is a redox-reaction
4Fe(s) + 3O
2
( g)
2Fe
2
O
3
( s)
Heat of Reaction
In almost all chemical reactions, heat is either
given off or absorbed
example: the combustion (oxidation) of carbon
liberates 94.0 kcal per mole of carbon oxidized
C(s) + O
2
(g) CO
2
(g) + 94.0 kcal

Heat of reaction: the heat given off or
absorbed in a chemical reaction
exothermic reaction: one that gives off heat
feels hot
endothermic reaction: one that absorbs heat
feels cold
Heat of Reaction
So, is this reaction exothermic or
endothermic?
C(s) + O
2
(g) CO
2
(g) + 94.0 kcal
Heat is released (on the product side)
Its Exothermic

How about:
2 NH
3
+ 22.0 kcal N
2
(g) + 3 H
2
(g)
Heat enters with reactant (on the reactant side)
Its Endothermic



What are Functions of NAD, NADP, FAD?

Electron carriers

Oxidation / reduction reactions

NAD and catabolic reactions
-- substrate oxidation
-- H- used for ATP synthesis

NADP and anabolic reactions
-- substrate reduction
-- e.g., --COOH to C=O to C-OH

Using NADH to make ATP


Electron transport and oxidative
phosphorylation
From glucose
Glycolysis ATP yield
____ NADH x ____ ATP _______
____ ATP _______
Transition Rx
____ NADH x ____ ATP x 2 pyr _______
Krebs cycle
____ NADH x ____ ATP x 2 pyr _______
____ FADH x ____ ATP x 2 pyr _______
____ GTP x ____ ATP x 2 pyr _______
Total 30
From a 12 carbon fatty acid
B-oxidation ATP yield
____ FADH x ____ ATP x 6 acCoA _______
____ NADH x ____ ATP x 6 acCoA _______
Krebs cycle
____ NADH x ____ ATP x 6 acCoA _______
____ FADH x ____ ATP x 6 acCoA _______
____ GTP x ____ ATP x 6 acCoA _______
Total 84
You should be able to complete a table to calculate energy yields from glucose or fatty acids (of any given length)
-- assuming 2.5 ATP per NADH (1.5 per glycolytic NADH) and 1.5 ATP per FADH
What are the biosynthetic roles
Of these pathways?
Complex designation Electron transport function

Complex I (NADH-Q reductase) oxidizes NADH to NAD
+
;
Iron containing flavoprotein transfers electrons to coenzyme Q

Complex II (Succinate-Q reductase) oxidizes succinate to fumarate;
FAD prosthetic group; SDH electron transfer to CoQ

Complex III (cytochrome reductase) reduces cytochrome C
Heme-iron cytochromes electron transfer CoQ to cyt C

Complex IV (cytochrome oxidase) oxidizes cytochrome C;
Copper and iron containing heme reduces O
2
to H
2
O
electron transfer cyt C to O
2
Table . Summary of redox complexes of the electron transport chain
CLINICAL CORRELATE RESPIRATORY CHAIN DEFECT
defects in each complex of the respiratory chain have been identified
associated with lactic acidemia because the high NADH
concentration favors the formation of lactate from pyruvate
blood lactate may be elevated 30-fold
blood pyruvate increases up to 10-fold
|NADH inhibition of TCA cycle + PDH |pyruvate
|pyruvate + |NADH |Lactate

ketones produced due to inhibition of TCA cycle and blood ratio of
|-hydroxybutyrate to acetoacetate increased in response to
increased mitochondrial NADH to NAD ratio
serum alanine is also increased due to decreased pyruvate
metabolism
FAD
FADH
2

e

DHAP
(2)
e

= electrons
CYTOPLASM
INNER
MEMBRANE
MATRIX
Glycerol-3-phosphate
dehydrogenase
G3P
Dihydroxyacetone
phosphate
(DHAP)
e

NAD
+
3-phosphate
Glycerol
e

(1)
CoQ
e

O
2

e

OUTER
MEMBRANE
Glycerol phosphate shuttle. Cytoplasmic glycerol 3-phosphate dehydrogenase
(1) oxidizes NADH. Glycerol 3-phosphate dehydrogenase in the inner membrane
(2) reduces FAD to FADH
2
.
NADH
Glucose
Pyruvate
GLYCOLYSIS
NAD
+
(6)
CYTOPLASM
e

= electrons
OUTER
MEMBRANE
(5)
MATRIX
Glu
Asp
(4)
INNER
MEMBRANE
KG
KG
Malate
(2) e
-

e
-

OAA NADH
NAD
+

(3)
e
-

Complex I
e
-

OAA
Malate
(1)
Glucose
Pyruvate
GLYCOLYSIS
e
-

NAD
+

NAD
+

NADH
e
-

The malate-aspartate shuttle.
Asp
Glu
ATP production
ATP
Synthase
NADH
NAD
+
I
n
n
e
r

m
i
t
o
c
h
o
n
d
r
i
a

m
e
m
b
r
a
n
e

Matrix
Matrix
Electron Transport Chain
Cytochrome B, Cytochrome
C, Fe-S proteins, etc.
e.g. in brown fats for heat
generation in small mammals.
Mitochondrion
has two
membrane
bilayers
Oxidative Phosphorylation takes place in
mitochondria for more ATP production
Glycolysis takes place in the cytoplasm; after glycolysis,
pyruvate is added with CoA using NAD+ to become Acetyl
CoA, CO2 and NADH.
Acetyl CoA is the fuel for Krebs Cycle to take place in the
matrix.
Oxidative phosphorylation depends on electron transfer and
the respiratory chain linking to TCA cycle create proton
gradient across the inner membrane of mitochondria.
The proton gradient powers the synthesis of ATP using ATP
Synthase
When these steps are blocked or uncoupled by uncoupling
proteins, no ATP made but only heat energy produced.
Review Questions
How does muscle produce ATP
(carbohydrates, fatty acids,
branched-chain amino acids)?
What are the key Ca
2+
regulated
steps?
Test and Expand your understanding about energy
1. How many phosphorylated intermediates are in the Krebs cycle?
Ans: None. GTP is synthesized from GDP + P
i
. GTP, however, is not a cycle
intermediate.
2. How is ATP generated in the Krebs cycle?
Ans: Indirectly. The reduced coenzymes, NADH and FADH
2
shuttle electrons to the
electron transport system and energy is preserved by ATP synthesis
3. Is pyruvate acetyl-CoA the only way to enter carbons into the Krebs cycle?
Ans: No. Any compound that can be converted into a Krebs cycle intermediate will
contribute carbons to the Krebs cycle. This applies to aspartate and glutamate,
which form OAA and o-ketoglutarate, respectively.
4. What numbers should I remember in order to calculate energy yield in the
Krebs cycle?
Ans: In terms of ATP, remember that each NADH is equivalent to 3, each FADH
2
to 2,
and each turn of the cycle 12 ATPs.
5. How many ATPs are generated when succinyl-CoA is oxidized in the cycle?
Ans: 30. One for GTP, two for FADH
2
and 3 for NADH must be added to the 24 for 2
turns of the cycle.