Unit 1 Introduction to Biomembranes

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UNIT 7

INTRODUCTION TO
BIOENERGETICS
“In every one of us there is a living process of combustion going on very similar to that
of a candle, and I must try to make that plain to you.”

—Michael Faraday, 1860

Structure
7.1 Introduction 7.5 Redox reactions

Expected Learning Outcomes Redox Reactions as Two

Half-reactions
7.2 Thermodynamics and Biological
Systems Redox Potential

Basic Terminology of Thermodynamics Nernst Equation

Laws of Thermodynamics and Their Standard Transformed

Significance Reduction Potential, E’°

7.3 Gibbs Energy Change Gibbs Energy and Reduction

Potential
Gibbs Energy Change and Spontaneity
7.6 Universal Electron
Gibbs Energy Change and Equilibrium
Carriers
Constant
7.7 Summary
Biochemical Standard State
7.8 Terminal Questions
Coupled Reactions
7.9 Answers
7.4 Non-equilibrium Thermodynamics
7.10 Suggested Readings

7.1 INTRODUCTION
All living organisms capture, transform, store and use energy to stay alive i.e.,
every living cell has the potential to derive energy from its environment; convert it
into a biologically useful form, and to utilize it for osmotic, chemical and
mechanical work needed for the sustenance of life. The study of the flow and
transformation of energy in biological systems is called bioenergetics and
forms the subject of this and the next few units. This, in fact is a part of a bigger
domain of study called ‘energetics’ which refers to the study of energy in terms 103
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of its flow and transformation within a system or between a system and its
surroundings. In simpler words, we can say that bioenergetics deals with the
generation of energy by harnessing the chemical energy of nutrients or light
energy from solar radiation (in phototrophs) and storage & utilization of energy
for performing different cellular functions. It also helps us understand myriads of
biochemical reactions interconnected through complex pathways in the cell.
We would begin the unit by recalling the basic terminology and the laws of
equilibrium thermodynamics about which you would have learnt in your earlier
classes. In this context, we would also recall thermodynamic functions viz.,
energy, enthalpy, and entropy and discuss their significance. Gibbs energy is
probably the most important concept required to understand bioenergetics. We
would give a detailed account of Gibbs energy, its importance as a criterion for
spontaneity, and its relationship with equilibrium constant. Thereafter, we
would discuss, in brief, about the need and relevance of ‘non-equilibrium
thermodynamics’ in the study of biological systems.
Further, as one of the important means of energy transfers in the living
systems is through redox reactions, so we would take up the basic aspects of
redox reactions. Herein we would talk about redox reactions, redox potential,
standard reduction potential and their significance. We would also introduce
the need for biochemical standard state, define it and discuss, how do the
parameters like Gibbs energy and redox potential change under biochemical
standard state and what is its significance. Towards the end of the unit, we
would take up universal electron carriers in terms of their role and significance
in harnessing and utilization of energy by biological systems. In the next unit
you would learn about ATP and other high energy compounds.

Expected Learning Outcomes

After studying this unit, you should be able to:

™ define bioenergetics and state its importance;

™ define and outline the significance of various thermodynamic functions;
™ state and explain the First and Second Laws of thermodynamics;
™ define spontaneity and state the criterion for spontaneity of biochemical
reactions;
™ define Gibbs energy and outline its significance;
™ relate Gibbs energy to the equilibrium constant;
™ state the need for biochemical standard state and define it;
™ explain coupling reactions and discuss their significance;
™ outline the need for non-equilibrium thermodynamics;
™ define oxidation and reduction;
™ explain the terms like standard reduction potential, cell potential and redox
potential and state their significance;
™ relate Gibbs energy with standard reduction potential; and
™ discuss the role of electron carriers in energy transductionin biological
systems.
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7.2 THERMODYNAMICS AND BIOLOGICAL
SYSTEMS
Thermodynamics is an experimental science based on certain generalisations
formulated on the basis of extensive observations of the universe we live in.
These generalisations are expressed in terms of laws of thermodynamics that
govern all physical, chemical, and biological processes. In order to understand
these laws and their application to biological systems, it is pertinent to recall
the basic terminology of thermodynamics.

7.2.1 Basic Terminology of Thermodynamics

Let us recall the basic terminology of thermodynamics about which you would
have learnt in your earlier classes.
System and its types: A ‘system’ in thermodynamics is defined as that part of
the universe, which is under consideration for the study. The system can be
large or small, simple or complex e.g., a single cell or the whole organism.
Further, a thermodynamic system must have well defined boundary that
separates it from the surroundings. The surroundingsin turn,are the
remaining (other than the system) part of the universe. The system and its
surroundings put together constitute the universe. However, for practical
purposes, the surroundings can be taken as that part of the universe with
which the system can interact.

If the boundary of the system is such that it allows the transfer of matter and All living organisms are
open systems as they
energy between the system and its surroundings through it, the system is do exchange matter
classified as open system. A glass test tube containing hot water is an (nutrients and waste
example of an open system as water can be removed or more water or some products) and energy
other kind of matter can be added to it and also the temperature of water can (as heat) with their
surroundings
be increased (by heating it) or decreased (by allowing it to cool down) by
suitable transfer of energy. A living cell is also an example of an open system
as there is a continuous flux of energy and matter to and from it.
If the boundary of the system does not permit the transfer of matter between the
system and its surroundings, but allows transfer of energy, it is called a closed
system. If the glass test tube containing hot water were suitably capped such
that the matter exchange is not allowed, it would become a closed system.
Further, a system having a boundary that does not permit the transfer of either
heat or matter to or from the surroundings is termed as an isolated system. If
the suitably capped glass test tube containing hot water is wrapped with a thick
thermally insulating material, it will become an isolated system. Different types
of thermodynamic systems are schematically shown in Fig.7.1.

Fig. 7.1: Schematic representation of different types of thermodynamic systems 105

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The thermodynamic State of a system: At any instant the system is in a thermodynamic state that
variables are also called can be defined by specifying the thermodynamic properties like, pressure, p;
state variables as these volume, V; temperature, T; amount, n; and density, U, etc. of the system. For
depend on the state of example, 50 cm3 of distilled water at 30oC in a beaker constitutes a state (the
the system and not on
the past history of the pressure here will be 1 atm.). If we change any of these variables of the
system. system we get a new state. For example, if we heat the water in the beaker to
70oC, we would get to a new state of the system. Further, the act of changing
the state of a system is called a thermodynamic process. In this example of
heating the water in a beaker, we have performed a thermodynamic process.

7.2.2 Laws of Thermodynamics and Their

Significance
As stated above, the laws of thermodynamics are generalisations that are
based on observations of the macroscopic world. These are physical laws of
nature that govern how the nature works. Contrary to the laws of society we
live in, there has not been any violation of the thermodynamic laws.

The first law of thermodynamics concerns conservation of energy i.e., in any

physical or chemical process the total energy of the universe (system +
surroundings) remains constant. In other words, the energy can neither be
created nor destroyed; it can only change forms. The first law is empirical in
nature and cannot be derived mathematically from more fundamental
principles. However, to understand it we need to recall the concept of internal
energy (U) of the system, which is the energy possessed by a given system
by virtue of its very existence and includes all possible types of energies,
which all the constituents of the system put together can have. The internal
The first law can also be energy of a system is a state function i.e., its value depends only on the state
stated, as “the energy of of the system and not on how it has been achieved. Further, it is important to
an isolated system is
note that the absolute value of the internal energy of any system cannot be
constant”
known, however the change in internal energy, 'U is measurable. If a system
exchanges some heat (q) with the surroundings and performs some work (w)
then we can mathematically express the First Law of thermodynamics as
'U = q + (-w) …(7.1)

As per the sign Where, 'U is the change in the internal energy of the system. The living cells
convention the heat are capable of interconverting different types of energies. The chemical energy
given to the system is stored in the food we consume is converted partly to high-energy bonds in ATP
positive and the heat
and other molecules and rest is released as heat. On a wider scale, we can
given by the system is
negative. Similarly the see that the solar energy (as sunlight) is converted to chemical energy during
work done on the photosynthesis, and this is utilized in the body for mechanical work (muscle
system is positive and contraction), chemical work (biosynthetic work) and osmotic work (movement
the work done by the
of molecules across the membranes).
system is negative.
Enthalpy is a property closely related to internal energy. It is defined as

H =U + pV …(7.2)
and corresponds to the heat exchanged by the system with the surroundings
under the conditions of constant pressure if the only work done is pressure-
volume work. However, if no gases are involved in the process of interest, say
a biochemical reaction then the change in volume is negligible and the
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enthalpy change is roughly same as the change in internal energy. That is why
in biological systems “energy” and “enthalpy” are generally used
interchangeably. A process (or a biochemical reaction) accompanied by
release of energy as heat to the surroundings is called an exothermic
process; whereas the one accompanied by absorption of energy as heat from
the surroundings is termed as an endothermic process.
The Second Law of thermodynamics concerns the spontaneity or the
directionality of a process. A spontaneous process is the one that occurs on
its own under a given set of conditions without the aid of any external agency.
For example, hot water in a container spontaneously cools down to the
temperature of its surroundings. The spontaneity of a process can be
ascertained in terms of another thermodynamic property called entropy which
literally means “a change within”. It is denoted as S, and is a measure of
randomness of the system; greater the randomness higher the entropy. An
ordered state (say a solid) is a low-entropy state, whereas a relatively
disordered state (say a liquid) is a high-entropy state. Thermodynamically,
entropy is defined in terms of the heat change and temperature of the system.
In a given reversible process (the one that occurs in an infinite number of very
small steps) the entropy change, dS, is given as the ratio of heat produced,
dq, and the temperature at which it is produced.
dqrev
dS = You may note that
T …(7.3) entropy provides a
description of energy
The entropy can also be visualized as a measure of dispersal of energy and
distribution, but it is not
matter, greater the dispersal higher the entropy. On a microscopic level the a representation of
entropy can be quantitatively interpreted in terms of the number of ways in energy that is available
which the molecules of a system can be arranged while keeping the total for useful purposes.
energy of the system constant.

S = k lnW …(7.4)
This number is called the number of microstates (W); a system with more
microstates has greater entropy.
Sum of the entropy
According to the Second Law of thermodynamics for a process to be change of a system and
spontaneous, the total entropy change for the system and surroundings its surroundings (or
combined entropy of the
('Ssys + 'Ssurr) or universe ('Stotal) must be positive (> 0) i.e., system and the
'Stotal = 'Ssys + 'Ssurr > 0 …(7.5) surroundings) always
increases for a
It is important to note that the entropy change for the system can be negative spontaneous process,
or positive. If for a spontaneous process the entropy decreases within a 'Stotal > 0
system, but it is accompanied by a greater increase in the entropy of the
surroundings. As a result, the total entropy change is positive and the process
is spontaneous. Thus, according to the Second Law, entropy change provides
a criterion for spontaneity of a process however; its direct application to
different systems is quite limited. This is so because it requires calculation of
entropy changes for the system as well as for the surroundings which are very
difficult. Gibbs energy change provides a relatively more convenient criterion
for spontaneity that depends only on the properties of the ‘system’. Let us
learn about the Gibbs energy and its significance. However, before that
answer the following simple questions to assess your learning. 107
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Define the types of thermodynamic systems giving suitable examples.

What are spontaneous reactions? What is the criterion for spontaneity of a

reaction?

7.3 GIBBS ENERGY CHANGE

Gibbs energy (G), named after American scientist Josiah Willard Gibbs,is a
measure of the net driving force of a process under the conditions of constant
Gibbs energy change,
temperature and pressure (which is applicable to most biological systems)
'G, is a quantitative
measure of the net and is defined as
driving force of a
process (at constant G = H–TS …(7.6)
temperature and
The Gibbs energy, like enthalpy and entropy, is a state function. Its absolute
pressure).
value for a system cannot be determined. However; it is possible to determine
the change in Gibbs energy in a process. It is given as

'G = 'H–T'S …(7.7)

The enthalpy change 'H, for a reaction equals the difference in the enthalpies
Biologists and
biochemists generally of the products and the reactants, whereas the entropy change 'S gives the
use Gibbs energy rather change in the system’s randomness.
than entropy in their
thermodynamic As you know, the enthalpy change essentially is a measure of the type and
calculations. number of bonds formed and broken in the reaction. It equals the amount of
energy exchanged as heat with the surroundings under conditions of constant
Thus, the change in temperature and pressure. However, all this energy is not available for useful
Gibbs energy is the work. Of this, a part of energy equal to T”S is dispersed to increase the
amount of enthalpy (or entropy, and is not available to do work, and the rest equal to 'G is the energy
heat) that is “free” to be available for useful work, which could be mechanical work, synthetic
converted into useful
work; the rest is involved (chemical) work or osmotic work. It is because of this fact that Gibbs energy
in entropy change. was earlier referred to as ‘Gibbs free energy’ or just ‘free energy’ however; this
term is no longer in use as lUPAC (the International Union of Pure and Applied
Chemistry) has recommended the term ‘free’ to be dropped.

It may be noted that heat can do work only when it goes from an object at
higher temperature to another at lower temperature. As the biochemical
reactions occur under conditions of constant temperature and pressure, the
cells cannot use heat as source of energy. The cells can only use Gibbs
energy, which they harness from nutrient molecules (in heterotrophs) or from
solar radiation (in phototrophs) and generate ATP molecules that act as the
energy currency of the cell. Gibbs energy acts as a criterion for spontaneity of
'G for a process refers a reaction, and also provides information about the equilibrium position.
to the amount of energy
that is available to 7.3.1 Gibbs Energy Change and Spontaneity
perform non-pV work.
Let us see how does change in Gibbs energy ('G) provide a criterion for
spontaneity. You would recall that as per the Second Law of thermodynamics
108 the total entropy change in a process must be positive, Eq. (7.5)
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'Stotal = 'Ssys + 'Ssurr > 0 …(7.5)

For a closed system under the conditions of constant temperature and Gibbs energy change
pressure, the enthalpy change can be equated with the heat flow across the ('G) expresses energy
exchange in terms of
walls of the system to the surroundings. The surroundings are much larger
spontaneity and the
than the system, and are under conditions of constant temperature and amount of useful energy.
pressure. Under such conditions, the heat transferred into the surroundings,
qsur, is equal to the change in the enthalpy of the system. We can, therefore,
write
'Hsys
'Ssurr =
T …(7.8)

Substituting in Eq. (7.5), we get

…(7.9) It is important to know

that though “G tells
Multiplying the equation by –T, and simplifying we get about the spontaneity of
a reaction but it gives no
T'Stotal = T'Ssys + 'Hsys < 0 …(7.10)
clue about the rate of
It implies that for a spontaneous process, the -T'Stotal term should be the reaction. The time
aspect falls in the
negative. As the temperature is always positive it is the same as saying that domain of chemical
the 'Stotal must be positive (Eq.7.5). Thus, in terms of the properties of system kinetics.
we can give the criterion for spontaneity as

'Hsys  T'Ssys < 0 …(7.11)

or 'Gsys < 0 …(7.12)
In other words, we can say that for a thermodynamic process to be
spontaneous its Gibbs energy must decrease or the change in Gibbs energy
should be negative (i.e., 'G< 0). You may note here that in Eq. (7.11) the
enthalpy change and entropy change, both are for the system only.
Chemical (or biochemical) reactions accompanied by a decrease in Gibbs
energy ('G < 0) are spontaneous and are called exergonic (Greek: ‘work
producing’), whereas the ones associated with an increase in the Gibbs
energy ('G > 0) are non-spontaneous and are called endergonic (Greek:
‘work consuming’) reactions. Schematic Gibbs energy profiles for endergonic
and exergonic reactions are given in Fig. (7.2).

Fig. 7.2: Schematic Gibbs energy profiles for a) endergonic and b) exergonic
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In exergonic reactions Gibbs energy becomes available for useful work,
whereas in endergonic reactions, we need to provide Gibbs energy for the
reaction to occur. The reactions in which 'G is equal to zero ('G = 0) implies
that these are at equilibrium. In other words, in such cases the rate of forward
reaction (formation of products) is equal to the rate of backward reaction
(formation of reactants) i.e., there is no net change in the concentrations of the
products and reactants.
Further, to use 'G as a criterion for spontaneity we need to consider the
temperature as well as changes in enthalpy and entropy of the system. A
reaction that is favorable from enthalpy point of view ('H <0) may not be
spontaneous if the entropy also decreases ('S <0) to an extent such that the -
T'S term becomes more than the decrease in enthalpy and 'G becomes
positive. On the other hand, an endothermic process ('H >0) can be
spontaneous if it is accompanied by a large increase in entropy. In such a
case -T'S term becomes responsible for making 'G negative. The interplay of
'H, 'S and T in deciding the spontaneity of a process is schematically
represented in Fig. 7.3.

Fig. 7.3: Schematic representation of dependence of spontaneity of a

reactionon 'H, 'S and T.

Having learnt about Gibbs energy change and its significance as a criterion for
spontaneity, answer the following simple question to assess your learning.

Define Gibbs energy and outline its significance.

7.3.2 Gibbs Energy Change and Equilibrium

Constant
We have stated above that the Gibbs energy change provides information
about the directionality of a reaction. To understand this, let us take a simple
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reaction in which a reactant A gets converted into product B and there is
equilibrium between A and B, which can be represented as

A B …(7.13)

At equilibrium, the concentrations of A and B do not change with time and are
related in terms of equilibrium constant, K

…(7.14)

The magnitude of K is a measure of the extent to which the reaction proceeds

in the forward direction. A large value of Kindicates that the equilibrium
mixture contains primarily the product B, whereas a small value of K suggests
the predominance of the reactant A in the reaction mixture. The ratio of the
concentrations of B to A at any instance, other than that at equilibrium, is
called mass action ratio or reaction quotient, Q. It has the same expression
as the equilibrium constant, the only difference is that the concentrations are
at any instance other than the equilibrium.

[B] …(7.15)
Q=
[A]
The value of reaction quotient ranges from 0 (corresponding to pure A) to
infinity (corresponding to pure B).For a general case in which ‘a’ moles of
substance A react with b moles of substance B to give c moles of substance
C and d moles of substance D as per the following expression:
aA + bB cC +dD …(7.16)
The expressions for the equilibrium constant and the reaction quotients would
respectively, be
[C]ceq [D]eq
d
Q= [C]c [D]d
Kc = and …(7.17)
[C]eq [D]eq Q= [C]c [D]d

The Gibbs energy change for the reaction given in Eq. (7. 16) is given as

'rG 'rGo +RT in Q …(7.18)

…(7.19)

Activity is ‘effective
Here, 'rGo is the standard Gibbs energy change, R and T being the concentration’. It can be
universal gas constant (8.314 J K-1mol-1) and absolute temperature in kelvin seen as the
(K), respectively. The quantities within the square brackets represent the concentration of the
concentrations (rather the activities, to be more precise), of the respective species corrected for its
non-ideal behavior at
products and reactants. The magnitude of Gibbs energy change ('rG) is a moderate or high
measure of the tendency of the system to move towards equilibrium. concentrations. In highly
dilute solutions the
You can note that 'rG depends on two quantities viz., 'rGo and RT ln Q. For a activity is same as
given reaction at temperature T, the value of 'rGo is fixed. However, the value concentration.
of Q in the second term varies according to the composition of the reaction
mixture. If the reaction is carried out under standard conditions i.e., the
concentrations (rather activities) of reactants and products being maintained
at unity (1M for solutions and one bar for gases) then the second term in
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Eq.(7.19) becomes zero (lnK= ln1=0), and

'rG = 'rGo …(7.20)

Thus, the standard Gibbs energy change refers to the Gibbs energy change
for a reaction in which all the reactants and products are at unit activity. In other
words it is a measure of the tendency of the system to move towards the
equilibrium when the system is at standard conditions. However, the actual
Gibbs energy change depends on the prevailing concentrations of the
reactants and products. It is important to note that the criterion for spontaneity
of a reversible reaction is 'rG < 0 and not 'rGo < 0.
The standard Gibbs
As stated above, at equilibrium, 'rG = 0 and Q= K; so we can write,
energy change ('rGo) can
be seen as an alternative 0 = 'rGo + RT In K …(7.21)
way of expressing the
equilibrium constant for 'rGo = - RT In K …(7.22)
the reaction.
This provides an important link between the thermodynamic quantities and
equilibrium constant. Substituting it back in Eq. (7.18) we get,

'rG = - RT In K + RT In Q …(7.23)
K K …(7.24)
'rG = - RT In = -2.303RT log
Q Q
Thus, if Q<K, 'rG is negative and the reaction is spontaneous, and
approaches the equilibrium where the 'rG becomes zero (as K=Q). On the
other hand, if Q >K, 'rG is positive and the reaction is spontaneous in the
reverse direction, and once again approaches the equilibrium where 'rG
becomes zero (as K=Q).

Let us take the reaction given in Eq. 7.13 again and start with pure A; the value
It is important to note of Q will be close to zero ('rG < 0), and the reaction would go in the forward
that the equilibrium direction, and some B will be formed. As more and more of B is formed, the
constant is related to the
value of Q goes on increasing and becomes lesser and lesser negative. Once
standard Gibbs energy
change ( 'rGo ) rather the equilibrium is established, Q becomes equal to K and 'rG becomes zero.
than to the actual Gibbs Similarly, if we start with pure B, the value of Q will be close to infinity ('rG > 0)
energy change ('rG) for for the reaction as written, and the reaction would go in the backward direction
the reaction.
(as the reverse reaction will be spontaneous now) and some A will be formed.
As more and more of A is formed, the value of Q goes on decreasing, and 'rG
becomes lesser and lesser positive. Once again when the equilibrium is
established Q becomes equal to K and 'rG becomes zero.
From this discussion, we can conclude that
x The Gibbs energy, G of the reaction is at its minimum at equilibrium, and it
increases as we go away from the equilibrium.

x Approach to the equilibrium is spontaneous from either direction

x Any fluctuation in the system at equilibrium will increase the Gibbs energy;
however, the system will spontaneously relax back to the equilibrium.

The Gibbs energy content of the reaction (A B) as a function of its

displacement from equilibrium for endergonic and exergonic reactions is
schematically shown in Fig. 7.4.
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Fig. 7.4: Gibbs energy content for the reaction, A B as a function of its
displacement from equilibrium for a) exergonic reaction and b) endergonic
reaction.

Now come back to the standard Gibbs energy change, 'rGo. It is equal to the
difference in the standard molar Gibbs energies of the products and the
reactants, i.e.,

'rGo Gmo (products)  Gmo (reactants) …(7.25)

The molar Gibbs energies of the reactants and products in turn can be given
in terms of their standard Gibbs energies of formation. So we can write

'rGo 'fGo (products)  'fGo (reactants) …(7.26)

As you know, the Gibbs energy of formation, 'fGo of a compound is defined as

the standard Gibbs energy change for the reaction involving formation of the
compound from its elements in their reference states, which are taken to be
zero.

7.3.3 Biochemical Standard State

It may be noted that two
In the context of defining standard Gibbs energy change we have defined the
different symbols are in
standard state for a reaction to be the one in which all the reactants and use for standard Gibbs
products are at unit activity (or 1M concentration for convenience). If the energy change as per
reaction involves H+ ions, the standard state would correspond to a pH=0. As biochemical standard
you know, biochemical reactions take place in the cell at a pH of about 7.0. In state. These are 'rGo’
such a situation, it would not be appropriate to use the standard Gibbs energy and 'rG’ o. However,
values based on the standard state (as given above) which is usually applied 'rG’ o is the one
recommended by
in physical chemistry. Therefore, there is a need to adopt a different standard
international committee
state for biochemical reactions. of chemists and
biochemists.
In case of biochemical reactions we use a standard state in which the
concentration of all the species are taken to be 1M as before, but the
concentration of H+ ions is taken to be 1.0 x 10-7M (i.e., pH = 7.0).In addition,
the concentration of H2O is taken to be constant as 55.5 M, and in the
reactions involving Mg2+ ions, their concentration is assumed to be constant
and taken as 1 mM = 1x10-3M, and the ionic strength, I=0.25. Further, in order
to make biochemical standard Gibbs energy to be distinct, it is given a
different symbol, 'rGco and is called standard transformed Gibbs energy
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change. For a given reaction, the standard transformed Gibbs energy change
('rGco ) is a constant, and is related to the equilibrium constant of the reaction
as per the following equation.

'rGco = -RTIn Kc …(7.27)

Here, Kc refers to the transformed equilibrium constant. Similarly, other

thermodynamic properties of the system based on biochemical standard state
Physical constants are called standard transformed constants e.g., 'rHc is called standard
based on the transformed enthalpy change and so on. Let us take an example to see the
biochemical standard
state are also called relationship between standard Gibbs energy change for a system using
standard transformed physical and biochemical standard state.
constants.
Let’s consider a general reaction in which one of the products happens to be a
hydrogen ion.

A+B C +H+ …(7.28)

For this reaction we can write the Gibbs energy change as

…(7.29)

Under conditions of physical standard state (all species at 1M concentration)

'rG 'rGo …(7.30)
Now, under the conditions of biochemical standard state the expression for the
Gibbs energy change becomes

…(7.31)

Equating, Eq. (7.30) and Eq. (7.31), we get

…(7.32)

Substituting the values of R and T, and converting natural log to the log to
thebase 10, we get

…(7.33)
If a reaction does not
involve water, hydrogen Solving, we get
ion or any ionisable
species then the …(7.34)
standard Gibbs energy
according to biochemical Similarly, for a biochemical reaction involving one of the reactants to be a
standard state is same hydrogen ion, we can show that
as that for physical
chemistry standard …(7.35)
state.
Similar to 'rGo being equal to the difference in the standard molar Gibbs
energies of the products and reactants, 'rGco is the difference in the standard
molar Gibbs energies of the products and reactants under biochemical
standard conditions. As discussed above, for 'rGo, in the present case 'rGco
gives the tendency of a system under biochemical standard state to move
towards the equilibrium. We have so far talked about three different versions of
the Gibbs energy change for a given reaction. It is worthwhile to summarise
them to understand differences between them.
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'rG : it is called Gibbs energy change for the reaction and equals the
difference between the Gibbs energies of the products and the reactants when
the system is not at equilibrium. It is the driving force behind the system going
to (attaining) the equilibrium.

'rGo : it is the standard Gibbs energy change of the reaction and equals the
Gibbs energy change of the reaction when all the reactants and products are
at physical chemistry standard state [concentration of 1 M or a partial
pressures of 101.3 kPa (for gases) and temperature of 298 K ]. It is the driving
force behind the system initially at standard state going to (attaining) the
equilibrium.

'rGco : it is the standard transformed Gibbs energy change of the reaction and
equals the Gibbs energy change of the reaction when all the reactants and
products are in their biochemical standard state [concentration of 1 M or a
partial pressures of 101.3 kPa (for gases), temperature of 298 K, [H+] =
1x 10-7M; [[H2O]=55 M], [Mg2+]= 1x 10-3 M. It is the driving force behind the
system initially at biochemical standard state going to the equilibrium.

Having learnt about biochemical standard state and the relationship between You may note that the
'G values are additive.
the standard Gibbs energies of reaction expressed in terms of physical and This in fact is a
biochemical standard states respectively, let’s take up coupling reactions, one consequence of Gibbs
of the important strategies used by the biological systems to transform energy being a state
chemical energy of the nutrients and light energy for their sustenance. function, which means
that its value for a
7.3.4 Coupled Reactions system depends on the
state of the system,
You know by now that in bioenergetics we deal with the way living cell gets irrespective of how it
has been achieved. In
energy from fuel metabolism or light capture and how it uses it for driving other words, whether
energy-requiring reactions. You also know that cellular functionality depends the state is obtained in
on the formation and activity of molecules like proteins and nucleic acids. The as single step or in a
formation of these ‘ordered’ macromolecules is energy intensive i.e., the cell number of steps.
needs energy ('G is positive) to synthesize them from smaller molecules. Further, the mechanism
The question remains, “How do these endergonic reactions occur?” of a reaction also has
no effect on 'G e.g.,
The answer lies in coupled reactions. In such reactions thermodynamically the 'G for the oxidation
of glucose is the same
unfavorable i.e., endergonic reactions are combined or coupled with energy
whether it occurs by
liberating i.e., exergonic reactions, such that the Gibbs energy change for combustion in a test
overall reaction is negative and the process is exergonic (spontaneous). The tube or by a series of
thermodynamic basis of coupling of endergonic and exergonic reactions is enzyme-catalyzed
that the two reactions share a common intermediate. That is, the product of
one reaction is a reactant in the other reaction. If, for an arbitrary case,
Reaction 1: A+B C 'rG’o =+5 kJ mol-1 …(7.36)

Reaction 2: C+D E 'rG’o =-12 kJ mol-1 …(7.37)

Net Reaction: A + B + D E 'rG’o =-7 kJ mol-1 …(7.38)
The Gibbs energy for the net reaction can be obtained by algebraic summation
of those of the individual coupled reactions. Let us understand it with the help
of a biochemical example.

Example1: The formation of adenosine triphosphate (ATP) from adenosine

diphosphate (ADP) involves a reaction of ADP with inorganic phosphate (Pi) 115
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as given below, Eq. (7.39). It is an non-spontaneous reaction ('G>0) and
would not occur without an input of energy.

ADP + PiÆ ATP + H2O 'rG’o =+30.5 kJ mol-1 …(7.39)

In a coupled reaction a In many of the metabolic pathways in the cell, ATP is produced by coupling the
spontaneous reaction reaction between ADP and Pi to another reaction that releases enough energy
provides the energy
(more than that required for formation of ATP) to make the coupled reaction
needed by a
subsequent non- spontaneous. In the glycolytic pathway (which you would have studied in your
spontaneous reaction. earlier classes) the formation of ATP is coupled with the conversion of 1,3-
bisphosphoglycerate into 3-phosphoglycerate, Eq. (7.40).

1,3-Bisphosphoglycerate + H2O
Æ3-phosphoglycerate + Pi; 'rG’o= 49.4 kJ mol-1 …(7.40)

The coupling between the two reactions is brought about by the enzyme,
phosphoglycerate kinase that catalyses both the reactions. The net reaction
can be obtained by combining Eq. (7.39) and Eq. (7.40) and simplifying as
follows
1,3-Bisphosphoglycerate + H2O Æ3-phosphoglycerate + Pi; 'rG’o= -49.4 kJ mol-1
ADP + PiÆ ATP + H2O 'rG’o =+30.5 kJ mol-1
1,3-Bisphosphoglycerate + ADP Æ3-phosphoglycerate + ATP'rG’o = 18.9 kJ

You may note here that the net reaction is spontaneous ('rG’o<0; exergonic).

The overall Gibbs energy change for the coupled reaction can be used to
calculate the equilibrium constant for the reaction. We can take one more
example of coupling reaction from glycolytic pathway to understand this.

Example 2: The very first step of glycolysis involves the conversion of glucose
to glucose-6-phosphate, again an endergonic reaction, Eq. (7.41).
Glucose+ Pi Æ glucose- 6- phosphate; 'rG’o= 13.8 kJ mol-1 …(7.41)
The required Gibbs energy is provided by hydrolysis of ATP,
ATP + H2O Æ ADP + Pi 'rG’o = -30.5 kJ mol-1 …(7.42)

The net reaction is

Glucose+ ATP + H2O Æglucose- 6- phosphate + ADP 'rG’o = -16.7 kJ mol-1
…(7.43)

This reaction is catalysed by the enzyme hexokinase that facilitates the

transfer of phosphate group from ATP to glucose. As you know that 'rG’o and
the equilibrium constant are related, we can calculate the equilibrium constant
for this reaction as follows.

'rG’o = - RT InK’ …(7.27)

Rearranging, we get
…(7.44)

Substituting the value of the standard transformed Gibbs energy change, R

116 and T (298K) we get, K´ = 846
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You may solve the following simple questions to assess your understanding of
transformed Gibbs energy change and coupled reactions.

Derive the relationship between the standard Gibbs energy changes for the
following reaction expressed in terms of physical and biochemical standard
states respectively.
A + nH+ B + C

What are coupled reactions? Discuss their importance giving a suitable

example.

7.4 NON-EQUILIBRIUM THERMODYNAMICS

As stated above, a living cell is an open system; there is a continuous flux of
energy and matter into and out of the cell. At a given instance it is in a steady
state i.e., the concentrations of different constituents in it remain constant over
a period of time. In other words, the input of energy (and matter) and output of
energy (and matter) is equal. As the continuous flow of matter across the
cellular boundaries does not allow the establishment of a true equilibrium, the
laws of thermodynamics (equilibrium thermodynamics to be specific) are not
completely applicable to the cell. Though under the conditions of constant
temperature and pressure, if we focus our attention on to a set of reactions in
the cell, the system is quite close to equilibrium and the thermodynamic laws
are fairly applicable. However, a better and detailed study of cellular reactions
requires a non-equilibrium thermodynamic treatment. This is a domain of
study, which was recognised more than a hundred years ago, and is still not
fully established. A detailed account of non-equilibrium thermodynamics is
beyond the scope of this course.
Oxidation-reduction (redox) reactions play very crucial role in the supply and
transformation of energy in biological systems. Let us recall the basic aspects
of redox reactions and learn about their roles.

7.5 REDOX REACTIONS

You would recall from your earlier studies that the oxidation involves the loss
of one or more electrons by an atom, molecule or an ion. In the following
reaction, the ferrous ion is getting oxidised to ferric ion by the loss of one
electron.

…(7.45)
Reduction, on the other hand, is the process that involves the gain of one or
more electrons by an atom, molecule or an ion. In the following reaction
chlorine molecule get reduced to chloride ion by gaining electrons
…(7.46)

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Further, you would also recall that the oxidation and reduction processes
always occur together in a reaction. In other words, the loss of electrons by
one substance is always accompanied by a gain of electron by some other
species, and consequently there are no free electrons in a chemical system.
Such reactions involving oxidation and reduction processes are termed redox
reactions or electron transfer reactions.

In a reaction, the species possessing higher affinity for electrons cause the
oxidation of other substances by abstracting electrons from them. These
The redox reactions
involve the loss of
species are called oxidising agents or oxidants. In the process, the oxidising
electrons by one agents themselves get reduced (as they gain electrons). In biological
chemical species, systems, oxidations can take place in a variety of ways, such as addition of
which is thereby oxygen, or removal of electrons or removal of hydrogen (dehydrogenation). Of
oxidized, and the gain
of electrons by another,
these, dehydrogenation is the most common form of biological oxidation. On
which is reduced the other hand, the species that readily give up electrons cause the reduction
of other substances. These are called reducing agents or reductants, and
in the process of reducing a substance they themselves get oxidised.
Work done in living systems depends on flow of electrons in oxidation-
reduction reactions occurring in them. For example, in heterotrophs food acts
as a source of electrons. The food molecules in the reduced form are oxidised
(e.g., glucose is oxidised to carbon dioxide and water), and the electrons
obtained in this process move through different metabolic intermediates to
dedicated electron carriers in enzyme-catalyzed reactions. In other words,
these organisms consume the organic materials produced by photosynthetic
organisms and use them as metabolic fuel. The electron carriers in turn,
donate these electrons to species with higher electron affinities. This is
accompanied by release of energy, which is used for performing different types
of work. You would learn about these electron carries in the next section. On
the other hand, the phototrophs use energy from light to produce chemical
energy in the form of nutrients like glucose which also involves redox
reactions.

7.5.1 Redox Reaction as Two Half-reactions

Continuing with the basic aspects of oxidation-reduction reactions, let us take
an example of a redox reaction. You are familiar with Benedict reaction used to
check for the presence of reducing sugars like glucose.

…(7.47)
In this reaction, the cupric ions are reduced to cuprous oxide by the aldehyde
group in glucose; which itself is oxidised to carboxylic acid group. Though both
oxidation and reduction occur simultaneously in redox reactions, it is
convenient to consider the oxidation and reduction reactions separately as two
half-reactions while describing electron transfers. In the above case, we can
write,
(Oxidation) …(7.48)

(Reduction) …(7.49)
The sum of the two half-reactions gives the whole reaction. This method of
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the concept of redox reactions. However, you may note here that this idea of
two half-reactions is for convenience of understanding; there are no free
electrons in a reaction. The electrons in these conceptual reactions can be
seen as being “in transit”. In general, these half-reactions can be represented
as conjugate redox pairs or redox couples,

Electron donor (1) Electron acceptor (1) + e– …(7.50)

Electron donor (2) + e– Electron acceptor (2) …(7.51)

Net reaction:
Electron donor (1) + Electron acceptor (2) Electron acceptor(1) +
Electron donor (2) …(7.52)
The redox couple of oxidized and reduced species in a half-reaction is
denoted as Oxidised form/Reduced form e.g., Fe 3+/Fe2+. Thus, in a redox
reaction, there would be two conjugate redox pairs and there would be
spontaneous electron transfer from the electron donor of one redox pair to the
electron acceptor of the other. Now, questions arise that in a redox reaction,
which species would act as an oxidising agent and which would be a reducing
agent? How do we determine that? Also, which of the two oxidising or reducing
agents is stronger, and why? Qualitatively speaking, a species possessing
stronger affinity for electrons cause the oxidation of other species by
abstracting electrons from it and act as oxidising agent. On the other hand,
the species that readily gives up electron act as reducing agent and cause the
reduction of other substances.

Quantitatively, the relative affinity of the electron acceptors of the redox pairs is
expressed in terms of standard reduction potential, E0, (in volts) of the
redox pair. That is, the potential of the reactions given in the reduction equation
to occur. Let us learn about the standard reduction potential of the redox pairs
and its significance.

7.5.2 Redox Potential

The reactions in the redox couples given above are equilibrium reactions. The
magnitude of their equilibrium constants can provide information about their
relative strength. However, handling electrons in such cases is an issue. The
situation is similar to the one in which a metal is dipped into a solution of its
ions. For example, if copper wire is dipped in a solution of copper ion (i.e.,
copper electrode) the following equilibrium exists
…(7.53)

Similarly, for zinc metal dipped in a solution of zinc ion (i.e., zinc electrode) the
following equilibrium exists
…(7.54)

These two electrodes can be combined to form an electrochemical cell

(Daniel cell). The relative strength of the electron acceptors (Cu2+ and
Zn2+ions) to accept the electrons would determine which of the two electron
acceptors would accept the electrons and which of the two electron donors
(Cu and Zn) would donate the electrons in the electrochemical cell. The
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potential difference between the two electrodes would depend on their
standard electrode potentials. How do we know the standard electrode
potentials?

The standard electrode potential of these electrodes can be obtained by

setting up cells of these two electrodes individually with reference hydrogen
electrode, which has been assigned a potential of 0.00 V at all temperatures.
A half-cell that takes
electrons from the The measured potential of the cell between a redox couple (under standard
standard hydrogen cell conditions; 25°C, solutes at 1 M concentration, gases at 101.3 kPa) and
is assigned a positive hydrogen electrode is called the standard reduction potential or standard
value of E0, and one
that donates electrons
redox potential. For zinc electrode,
to the hydrogen cell, a
⇌ …(7.55)
negative value.

⇌ …(7.56)

You would recall from your earlier studies that the combination of a given
electrode and hydrogen electrode is represented as following cell diagram.

…(7.57)

As a matter of convention, the standard hydrogen electrode is always taken as

the electrode on the left of the cell diagram and the electrode under
consideration on the right. The standard cell potential is always taken as

…(7.58)

Subtracting Eq. (7.56) from Eq. (7.55) we get the reaction as

…(7.59)

The measured value of the cell potential for cell reaction given in Eq. (7.59) is
found to be -0.77V. A negative value of Eo means that in this cell, zinc metal
would get oxidised to Zn2+ ions (i.e., zinc electrode would act as anode)
whereas hydrogen ions would get reduced to hydrogen gas i.e., the following
(and not the one given in Eq. (7.59)) reaction would occur spontaneously.

…(7.60)

Now if we replace the zinc electrode with copper electrode (under standard
conditions) and measure the electrode potential we get a value of 0.34 V. This
indicates that in this cell Cu2+ ions would get reduced to copper metal (i.e.,
copper electrode would act as cathode) whereas hydrogen gas would get
oxidised to hydrogen ions (i.e., hydrogen electrode would act as anode) i.e.,
the following reaction would occur.

…(7.61)

Different redox couples are arranged in terms of decreasing value of their

standard reduction potentials as given in Table 7.1.

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Table 7.1:Standard reduction potentials (Eo) for some redox half-
reactions at 298 K
Reduction half-reactions Eo, Volts

Oxidising agent Reducing agent

+2.65

+2.01

+1.23

⇌ +1.07

+0.77

+0.34

0.00

-0.25

Zn(s) -0.77

-1.66

-2.38

-3.04

In the event of an electrochemical cell constructed by combining any

two electrodes; the electrode with a higher value of Eo would act as You may note here that
the standard redox
cathode and the one with a lower value would act as anode. potentials do not give
Going further, if we make a cell by combining zinc electrode and copper any information about
the speed of the
electrode (i.e., make Daniel cell); there would be oxidation at the zinc reaction.
electrode (anode, lower Eo) and reduction at the copper electrode (cathode,
higher E0). The cell would be represented as
…(7.62)

The standard cell potential for the cell can be obtained by using Eq. (7.58).

…(7.63)
Having learnt about redox reactions and redox potential why don’t you assess
your understanding by solving the following simple question?

NADH can reduce pyruvate to lactate by the following reaction catalysed by

lactate dehydrogenase.

Write this redox reaction in terms of two half-cells. 121

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7.2.1 Nernst Equation
The discussion above has been on the standard reduction potential i.e., the
reduction potential of a redox couple when all the species are at standard
conditions (1 M concentrations). However, in real systems, the actual potential
of the redox couple would depend on the standard reduction potential and the
actual concentrations of the species involved. The relationship between the
actual reduction potential and standard reduction potential for a given redox
couple was formulated by Walther Nernst, and is expressed in terms of an
equation called Nernst equation according to which, for the electrode
reaction:

…(7.64)

The reduction potential of the electrode at any concentration measured with

respect to standard hydrogen electrode is given by the following equation

…(7.65)
o
Where E n+ is the standard electrode potential, R is the gas constant, T is
M /M
the absolute temperature, n is the number of electrons participating in the
reaction, and F is the Faraday’s constant. The logarithm term is the ratio of the
concentrations of the reduced and oxidised forms of the species. As the
concentration (rather activities) for pure solids and liquids are taken as 1, the
equation becomes,
 …(7.66)

After converting natural logarithms to logarithm to the base 10, substituting the
numerical values for the constants, and assuming the temperature as 298K,
we can write the equation as given below.

 …(7.67)

For copper electrode, the Nernst equation at 298 K would be

 …(7.68)

This is about one electrode or half-cell. In order to write the Nernst equation for
the cell, say Daniel cell, we would proceed as follows.

Anode: …(7.69)

Cathode:  …(7.68)

The cell potential,

…(7.69)

…(7.70)

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…(7.71)

…(7.72)

Having learnt about the basic aspects of redox reactions in general, let us now
take these up in the context of biochemical systems.

7.2.2 Standard Transformed Reduction Potential, E1°

You have learnt above that the standard reduction potential refers to the
reduction potential of a redox couple under standard conditions. Here,
standard conditions refer to the physical chemistry standard state
(concentration/activities=1, p=1 bar, T=298 K and so on). However, as
discussed in the context of Gibbs energy for biochemical systems, we need to
consider biochemical standard state. Accordingly, the standard reduction
potential of a redox couple gets modified, and is called standard transformed
reduction potential. Let’s take an example of a biochemical redox couple
(fumaric acid/succinic acid) involving hydrogen ions to understand it
…(7.73)
The Nernst equation for the couple for certain concentrations of fumaric acid
and succinic acid can be written as

…(7.74)

Under conditions of physical standard state the concentration (or activity) of

hydrogen ions would be 1. Substituting we get,

 …(7.75)

However, under biochemical conditions the concentration of hydrogen ions

would be different from 1, and the potential would also be different. We can
write,

 …(7.76)


…(7.77)


…(7.78)

The first and the second terms on the right hand side equals .
Substituting, we get

E Fum/Succ E´fum/succ  0.059 pH …(7.79) 123

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Thus, for fumaric acid/succinic acid redox couple, the potential depends on the
pH of the medium. However, if the succinic acid and fumaric acids are at unit
concentrations, and pH =7.0 (under biochemical standard state), then the Eq.
(7.79) becomes
o
 Eofum/succ
E´Fum/Succ  0.059 u 7 …(7.80)
That is, the transformed standard reduction potential for the fumaric acid/
succinic acid redox couple would be lesser suggesting lower tendency for
fumaric acid to get reduced.
o
 Eofum/succ
E´Fum/Succ  0.413 V …(7.81)
This fact can also be rationalised as the effect of decreasing the concentration
of hydrogen ions (increasing the pH) on the equilibrium given in Eq. (7.73). The
equilibrium would shift to the left i.e., the tendency of fumaric acid to get
reduced decreases. The standard transformed reduction potentials of some
biochemically important redox couples are given in Table 7.2.

Table 7.2: Standard transformed reduction potentials of some

biochemically important redox couples

Reduction half-reaction E’o/ V

Oxidizing agent Reducing agent
–
O2 + 4H+ + 4e Æ 2H2O + 0.81
–
Fe3 + (Cyt f) +e Æ Fe2 + (Cyt f) +0.36
–
O2 + 2H2O + 4e Æ 2H2O2 + 0.30
–
Fe3 + (Cyt c) +e Å Fe2 + (Cyt c) + 0.25
–
Fe3 + (Cyt b) +e Æ Fe2 + (Cyt b) + 0.08
–
Dehydroascarbic acid + 2 H+ + 2e Æ Ascorbic acid + 0.08
–
Coenzyme Q + 2 H+ + 2e Æ Coenzyme QH2 + 0.04
–
Oxaloacetate2– + 2 H+ + 2e Æ Malate2– - 0.17
–
Pyruvate– + 2H+ + 2e Æ Lactate– - 0.18
–
FAD + 2 H+ + 2e Æ FADH2 - 0.22
–
Glutathione (ox) + 2 H+ + 2e Æ Glutathione (red) - 0.23
–
Lipoic acid (ox) + 2 H+ + 2e Æ Lipoic acid (red) - 0.29
–
NAD+ + H+ + 2e Æ NADH - 0.32
–
2H2O + 2e Æ H2 + 2 OH– - 0.42
–
Ferredoxin (ox) +e Æ Ferredoxin (red) - 0.43

Æ O2 - 0.45

Strongly reducing

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7.5.5 Gibbs Energy Change and Reduction Potential
We have seen above that the magnitude and the sign of the reduction poten-
tials indicate the direction in which a redox reaction would proceed. Also, we
are aware that the Gibbs energy change for a reaction also provides similar
information. The question arises, "Are these two related?" The answer is 'yes',
and in fact, it is a very significant relationship. The Gibbs energy change and
the standard reduction potential change for a redox reaction are related as
follows.
'G nF'E …(7.82)

'G´O nF'E´O …(7.83)

Where n is the number of electrons transferred in the reaction, and is the

difference in reduction potentials of the two half-cells as

…(7.84)
That is, we subtract the standard reduction potential of the half-cell undergoing
oxidation from that of the half-cell undergoing reduction. In case of biochemical
systems, we use the corresponding transformed reduction potentials.

Let us recall the example of fumaric acid/ succinic acid redox couple and
rewrite the Eq. (7.81)
…(7.85)
Multiplying throughout by -2F, we get

…(7.86)
This gives

…(7.87)

Thus, the Gibbs energy change for the reaction under biochemical conditions
(pH=7.0) is lesser negative, which means the reaction is more spontaneous at
pH 0 than at pH 7.0.The relationships given in Eq. (7.82) and (7.83) can be
conveniently used to determine the Gibbs energy change for a redox reaction,
and hence calculate the corresponding equilibrium constant. Let us take an
example.
Example 3: In the terminal respiratory chain, the conversion of NADH to NAD+
generates two electrons, which are used to reduce molecular oxygen to water.
The corresponding half-cell reactions are
⇌

The overall reaction is

The value of 'E’O can be calculated as

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From Eq. (7.83) we know that,

Substituting the values, we get

A part of this large amount of Gibbs energy is used for the synthesis of ATP
molecules.

Having learnt about redox reactions, redox couples and the related concepts
we are equipped to take up the universal electron carriers. However, before
moving ahead, answer the following simple questions.

The half-reaction of the NAD+/NADH redox couple is given as under

Write the Nernst equation for the same.

7.6 UNIVERSAL ELECTRON CARRIERS

As discussed above, the redox reactions are accompanied by a change in
Gibbs energy. Further, the magnitude and sign of the standard reduction poten-
tial (or transformed standard reduction potential) determine the magnitude and
sign of the Gibbs energy change, and this in turn dictates the spontaneity of the
reaction and the equilibrium constant. A great deal of energy transfer in biologi-
cal systems happens through redox reactions, and is facilitated by electron
carriers. It is, therefore, pertinent to learn about the electron carriers and their
role in biological systems. The electron carriers like, NAD+, NADP, FMN, and
FAD undergo reversible oxidation and reduction in a number of the electron-
transfer reactions in metabolism.
In the catabolic pathways, the energy released during the oxidation of nutrients
is picked up by the oxidised electron carriers, which in turn get reduced. The
energy is captured in terms of the electrons in the reduced form of the electron
carrier. This energy becomes available when these electrons are eventually
transferred to oxygen in the electron transport chain. Let us take the example of
oxidation of glucose to understand the role of electron carriers.

You know that the oxidation of glucose is the source of energy in many organ-
isms. The aerobic oxidation of glucose to carbon dioxide and water can be
represented as

…(7.88)
A large amount of Gibbs energy is released in this process; the value of trans-
formed Gibbs energy change 'G´O being 2840 kJ/mol. The overall reaction
involves the oxidation of glucose and the reduction of oxygen. The correspond-
ing half-reactions can be given as

⇌ …(7.89)

…(7.90)
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This means that a total of 24 electrons are transferred to 6 oxygen molecules.
This transfer of electrons does not happen directly to oxygen in one step;
instead these are transferred to the coenzymes NAD+ and FAD aided by a
number of enzymes of glycolytic pathway and Kreb's cycle to form 10 NADH
and FADH2 molecules. These two reduced coenzymes viz., NADH and FADH 2
carry these electrons and transfer them to oxygen in a meticulously organized
set of reactions in the electron transport chain. The transferred electrons
participate in a series of redox reactions in four enzyme complexes before
reducing O2 to H2O. In the process of transferring the electrons to other spe-
cies, the reduced coenzymes (NADH and FADH2) are reoxidised to NAD+ and
FAD and become available for other reactions of substrate oxidation. You
would learn in details about the electron transport in Unit 9 and 11. Let us begin
with NADH. You may recall from example 3 that NAD+ (the oxidised form of
NADH) gets reduced by taking up a proton and two electrons.

These electrons are eventually transferred to oxygen and the Gibbs energy
change for the overall process is. This Gibbs energy is harnessed in a
stepwise manner as given in figure in the margin. Each of these three steps
contributes to the synthesis of ATP by oxidative phosphorylation. Overall
oxidation of one molecule of NADH results in the synthesis of approximately
2.5 molecules of ATP. NADH plays the role of electron carrier in a number of
reactions. In general, the reaction of NAD+ with the substrate (AH2) may be
given as:

…(7.91)
For example, the conversion of alcohol to aldehyde by alcohol dehydrogenase
can be given as

⇌ …(7.92)
On the other hand, the reduced coenzyme, FADH2 gets oxidised to FAD by
Coenzyme Q (CoQ).

…(7.93)

The transformed standard Gibbs energy change for the process is -16.5 kJ
mol-1. This is not sufficient to synthesise ATP as it requires much more Gibbs
energy. The function of this reaction is just to inject the electrons from FADH2
into the electron transport chain. The details of the processing of electrons in
the electron transport chain and their eventual transfer to oxygen would be
taken up in Unit 9. For now let us sum up what we have discussed in this unit. 127
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7.7 SUMMARY
Let us recapitulate what we have learnt so far:
z The study of the flow and transformation of energy in biological systems is
called bioenergetics and in fact is a part of a bigger domain of study called
'energetics' which refers to the study of energy in terms of its flow and
transformation within a system or between a system and its surroundings.
z A 'system' in thermodynamics is defined as that part of the universe, which
is under consideration for the study, while the 'surrounding' in turn, is the
remaining (other than the system) part of the universe. The system and its
surroundings put together constitute the universe.
z Open system is such that it allows the transfer of matter and energy
between the system and its surroundings through it, while closed system
does not permit the transfer of matter but allows transfer of energy.
z The first law of thermodynamics concerns conservation of energy i.e., in
any physical or chemical process the total energy of the universe (system +
surroundings) remains constant.
z The Second Law of thermodynamics concerns the spontaneity or the
directionality of a process. A spontaneous process is the one that occurs on
its own under a given set of conditions without the aid of any external
agency.
z Gibbs energy (G), named after American scientist Josiah Willard Gibbs, is a
measure of the net driving force of a process under the conditions of
constant temperature and pressure.
z Coupling reactions means reactions which are thermodynamically
unfavorable (i.e., endergonic reactions) are combined or coupled with energy
liberating (i.e., exergonic reactions), such that the Gibbs energy change for
overall reaction is negative and the process is exergonic (spontaneous).
z Work done in living systems depends on flow of electrons in oxidation-
reduction reactions occurring in them.
z Standard reduction potential or standard redox potential is the potential of
the cell measured between a redox couple (under standard conditions;
25°C, solutes at 1 M concentration, gases at 101.3 kPa) and hydrogen
electrode.
z The electron carriers like, NAD+, NADP, FMN, and FAD undergo reversible
oxidation and reduction in a number of the electron-transfer reactions in
metabolism. Two reduced coenzymes viz., NADH and FADH2 carry these
electrons and transfer them to oxygen in a meticulously organized set of
reactions in the electron transport chain. This plays a significance role in
harnessing and utilization of energy by biological systems.

7.8 TERMINAL QUESTIONS

1) What are endergonic and exergonic reactions?

2) Why do we need to define a separate standard state for biochemical

128 reactions? What is biochemical standard state?
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3) Outline the need for non-equilibrium thermodynamics.

4) The biochemical reaction for hydrolysis of ATP is given below

Calculate the Gibbs energy change for hydrolysis of ATP in a celI having the
concentrations of ATP, ADP, and Pi as 3.4 mM, 1.3 mM, and 4.8 mM,
respectively.

5) Standard Transformed Gibbs Energies of Hydrolysis of Creatine phosphate

and ATP are given as under.

Calculate the transformed equilibrium constant for the reaction involving

transfer of phosphate from Creatine phosphate to ADP.
6) Universal electron carrier, NADH is oxidised by oxygen as per the following
equation.

Write this redox reaction in terms of two half-cells.

7) Calculate the transformed standard Gibbs energy change, 'G´°, for the
following reaction involving reduction of acetaldehyde to ethanol by NADH

The E’ ° values for the NAD+/NADH and CH3CHO /CH3CH2OH redox

couples are given as -0.320 V and -0.197 V respectively.

7.9 ANSWERS
Self-Assessment Questions
1) A thermodynamic system that allows the exchange of energy as well as
matter with the surroundings is called an open system, for example, food
being cooked in an open vessel. On the other hand, if the system permits
the exchange of energy but not the matter it is termed as closed system,
e.g., cooked food kept in a closed metallic container. The isolated system
does not permit the exchange of either the energy or the matter. The food in
an insulated closed container is an example of an isolated system.

2) A spontaneous process is the one that occurs on its own under a given set
of conditions without the aid of any external agency. According to the Sec-
ond Law of thermodynamics for a process to be spontaneous, the total
entropy change for the system and surroundings ('Ssys + 'Ssurr) or uni-
verse ('Stotal) must be positive i.e., 'Stotal = 'Ssys + 'Ssurr >0

3) Gibbs energy (G), is defined as G = H -TS. It is a measure of the net driving

force of a process under the conditions of constant temperature and
pressure and provides a criterion for spontaneity of a reaction.
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4) The Gibbs energy change for reaction

Can be given as

Under conditions of physical standard state (all species at 1M

concentration)

'rG = 'rGo …(A)

Now, under the conditions of biochemical standard state the expression for
the Gibbs energy change becomes

…(B)

Equating, Eq. (A) and Eq. (B), we get

Substituting the values of R and T, and converting natural log to the log to
the base 10, we get

Solving, we get

5) The coupled reactions refer to the coupling / combination of an endergonic

reaction with an exergonic reaction such that the overall reaction is
exergonic i.e., spontaneous. This is mediated through a common
intermediate between the two reactions, and is a consequence of the fact
that the Gibbs energy is a state function. As an example, we can take the
formation of ATP by oxidative phosphorylation from Creatine phosphate as
per the coupling of following reactions

The overall reaction

is exergonic, and hence spontaneous.

6) To express a reaction as the difference of two reduction half-cells, first we

identify the species undergoing reduction and the product of reduction. Here
we see that pyruvate is getting reduced to lactate. The corresponding
reduction half-reaction can be written (from Table 7.2) as

Pyruvate + 2H+ 2e– Lactate

In the next step we subtract this reaction from the overall reaction to get

NADH – H+ – 2e– NAD+

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This can be rearranged to give

This is the oxidation half-reaction, which can be reversed to give the corre-
sponding reduction half-reaction

Thus the two half-reactions are as under

Pyruvate + 2H+ 2e– Lactate

7) The Nernst equation for the given half-reaction of the NAD+/NADH redox
couple would be as follows

Terminal Questions
1) Chemical (or biochemical) reactions accompanied by a decrease in Gibbs
energy ('G < 0) are spontaneous and are called exergonic, whereas the
ones associated with an increase in the Gibbs energy ('G > 0) are non-
spontaneous and are called endergonic reactions.

2) The physical chemistry standard state for a reaction is defined as the one
in which all the reactants and products are at unit activity (or 1M concentra-
tion). For the reaction involving H+ ions, this corresponds to a pH=0. How-
ever, as the biochemical reactions in the cell take place at a pH of about
7.0, the physical chemistry standard state is not appropriate. We need to
adopt a different standard state for biochemical reactions.

According to the biochemical standard state, the concentration of all the

species are taken to be 1M as in physical chemistry standard state, but the
concentration of H+ ions is taken to be 1.0 x 10-7 M (i.e., pH = 7.0). In
addition, the concentration of H2O is taken to be constant as 55.5 M and in
the reactions involving Mg2+ ions, their concentration is assumed to be
constant and taken as 1 mM = 1x10-3M, and the ionic strength, I=0.25.
3) A living cell is an open system having a continuous flux of energy and
matter into and out of it. At a given instance the cell is in a steady state i.e.,
the concentrations of different constituents in it remain constant over a
period of time. However, this continuous flow of matter across the cellular
boundaries does not allow the establishment of a true equilibrium. The laws
of thermodynamics, which are for systems under equilibrium, are not
completely applicable to the cell though they can provide certain details of
the systems. Therefore, there is a need for non-equilibrium thermodynamic
treatment for a better and detailed study/understanding of cellular reactions.

4) The Gibbs energy change for hydrolysis of ATP can be given as

It may be noted here that concentration of H2O does not appear in the
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expression as according to the biochemical standard state, it is constant
and is included in the 'rG’ o term.

Now substituting the values of different terms, we get,

[1.3u103] [4.8u103]
'rG 37700 Jmol1  (8.314 JK1mol1)(298K)(2.303)log
[3.4u103]

5) The reaction representing transfer of phosphate from Creatine phosphate to

ADP can be given as

This reaction can be obtained by adding reaction 1 and the reverse of

reaction 2 as follows

⇌
The corresponding Gibbs energy change would be

We know that: 'rG’ o = – RT In K’

Rearranging and simplifying:

Substituting the values we get, K’ = 4.63

6) In this reaction acetaldehyde takes two electrons from NADH, i.e., n=2. We
know that the transformed Gibbs energy and reduction potentials are
related as
'G’ o = – nF 'E’o
substituting the values of n, F and E'o, we get

'G’ o = –2(96500 J V–1mol–1) (0.123 V)

'G’ o = –23.7 kJ mol–1

7) In this reaction NADH is getting oxidised to NAD+ whereas CH3CHO is

getting reduced to CH3CH2OH. The corresponding reduction reactions are
as under

The 'E´ o value can be obtained by using Eq. 7.84 as

'E´ o = E´ o (reduction) – E´ o (oxidation)

Substituting the given values of transformed standard reduction potentials

we get
'E´ o = – 0.197 – (– 0.320) V = 0.123 V
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o o
We know that 'G´ = –nF'E´

Here, n=2; substituting the values, we get

'G´O 2 u (96500 Cmol1)(0.123 V) 23.70 kJmol1

7.10 SUGGESTED READINGS

z Garret, R.H., Grisham, C.M. (2016). Biochemistry (6th ed.). Boston,
Cengage Learning. ISBN-10: 1133106293, ISBN-13: 978-1133106296
z Berg, J.M., Tymoczko, J.L. and Stryer L., (2012) W.H. Biochemistry (7th
ed.), Freeman and Company (New York), ISBN:10: 1-4292-2936-5,
ISBN:13:978-1-4292-2936-4.
z Nelson, D.L., Cox, M.M. (2017). Lehninger: Principles of Biochemistry (7th
ed.). New York, WH: Freeman and Company. ISBN: 13: 978-1-4641-2611-6
/ ISBN:10:1-6412611-9.
z Lodish, H., Berk, A., Kaiser, C.A., Krieger, M., Bretscher, A., Ploegh, H.,
Amon, A., Scott, M.P. (2016). Molecular Cell Biology (8th ed.). New York,
WH: Freeman & Company. ISBN-13: 978-1-4641-0981-2.

z Voet, D.J., Voet, J.G., Pratt, C.W. (2008). Principles of Biochemistry (3rd
ed.). New York, John Wiley & Sons, Inc. ISBN:13: 978-0470-23396-2

133