BIOENERGETICS AND BIOLOGICAL OXIDATION

Bioenergetics is the study of energy transformation (processing and consumption) within the biological system.
It examines the changes in the energy content and its distribution when the body passes from defined state into a
thermal state at equilibrium. It is concerned with the initial and final states of biomolecule and can be used to predict the
direction and extent to which specific biochemical reaction can proceed.
Thermodynamic is concerned with heat and energy transformation occurring in the universe composed of a
system and its surrounding. Hence bioenergetics can be defined as the thermodynamics of a biological system.
A system may be defined as an entire organism or a single cell or reaction taking place inside a flask etc. In
general, a system may be regarded as open or close system. In an open system, matter and energy are expended. On
the other hand, a close system permits only the exchange of energy between it and its surrounding.

Surrounding
flask

Living systems are open because they consume nutrient (matter) from their surrounding and release waste product
(matter) and energy into the surrounding.
Several thermodynamic properties are defined as state function- enthalpy, entropy and free energy (Gibb’s free
energy). What does this means? (What is a state function?) It means that its value does not depend on the pathway
used to synthesize a specific substance e.g. the energy content of glucose molecule does not depend on how it was
made (either through photosynthetic pathway, hydrolysis of maltose or lactose). On the other hand, entities such as
work and heat are not state function because it refers to the energy changes occurring in a process. Thus, they depend
on the pathway used. For example, living organism oxidizes glucose and use part of the energy to perform cellular work
such as muscle contraction or transport of solutes. If the glucose is oxidized in different pathway e.g digestion in a
laboratory dish, the energy of the glucose must be transformed into heat with no measurable work done. The energy
content of glucose will be the same, however, the work done is different.
Knowledge of these state functions enables biochemist to predict whether a reaction would occur
spontaneously or not. This is so because the direction and extent to which a reaction proceeds are determined by
change in the enthalpy, entropy and free energy.

Enthalpy refers to the total heat content while entropy refers to the degree of randomness or disorder. Free energy
refers to the energy available for doing useful work. Enthalpy and entropy are related to the first and second laws of
thermodynamics respectively. Free energy is derived from a mathematical relationship between enthalpy and entropy
viz visa.
∆G = ∆H - T∆S where

∆G= change in Gibb’s free

∆H= change in enthalpy
∆S= change in entropy
T= absolute temperature in Kelvin

Laws of Thermodynamics
All processes that occur in the universe are subject to the basic laws of thermodynamics. The reactions that
occur in living cells are no exception.
First Law of Thermodynamics: The total energy of the universe does not change. This does not mean that the form of
the energy cannot change. Simply put; energy can neither be created nor destroyed. In any given process, one form of
energy may be converted into another but the total energy of the system plus its surroundings remains constant.
Nothing is said about the relative usefulness of different forms of energy or the direction of a process or reaction.
Indeed, chemical energies of a molecule can be converted to thermal, electrical or mechanical energies. The
internal energy of a system can change only by work or heat exchanges. From this the change in the free energy of a
system can be shown by the following equation:
ΔE = q – w
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 Where ΔE = change in the internal energy, q= heat absorbed by the system, w= work done by the system.
When q is negative heat has flowed from the system and when q is positive heat has been absorbed by the system.
Conversely when w is negative work has been done on the system by the surrounding and when positive, work has
been done by the system on the surroundings.
In a reaction carried out at constant volume no work will be done on or by the system, only heat will be
transferred from the system to the surroundings. The end result is that:
ΔE = q
When the same reaction is performed at constant pressure, the reaction vessel will do work on the surroundings. In this
case:
ΔE = q – w
where
w = PΔV

Note: w = PΔV + VΔP

where P and V are changes in pressure and volume respectively.
If the process takes place at constant pressure and volume, then
ΔP = 0
ΔV= 0
ie the workdone = 0 + 0 = 0 consequently the equation becomes ΔE = q

When the initial and final temperatures are essentially equal (e.g. in the case of biological systems):
ΔV = Δn[RT/P]
therefore,
w = ΔnRT
By rearrangement of the above equations one can calculate the amount of heat released under constant pressure:
q = ΔE + w = ΔE + PΔV = ΔE + ΔnRT
In this last equation, Δn is the change in moles of gas per mole of substance oxidized (or reacted), R is the gas constant
and T is absolute temperature.

Enthalpy
Since all biological reactions take place at constant pressure and temperature the state function of reactions
defined to account for the heat evolved (or absorbed) by a system is enthalpy given the symbol, H. The changes in
enthalpy are related to changes in free energy by the following equation:
ΔH = ΔE + PΔV
The equation is written in this form because we are addressing the constant pressure situation. In the biological setting
most of all reaction occurs in a large excess of fluid, therefore, essentially no gases are formed during the course of the
reaction. This means that the value ΔV, is extremely small and thus the product PΔV is very small as well. The values
ΔE and ΔH are very nearly equivalent in biological reactions.
Stated above was the fact that state functions, like ΔH and ΔE, do not depend on the path taken during a
reaction. These functions pertain only to the differences between the initial and final states of a reaction. However, heat
(q) and work (w) are not state functions and their values are affected by the pathway taken.
Reversible and Irreversible Reactions
In an idealized irreversible reaction such as one done by expanding an ideal gas against zero pressure, no work
will be done by or on the system so the:
w=0
In the case of an ideal gas (whose molecules do not interact) there will be no change in internal energy either so:
ΔE = 0
since ΔE = q – w, in this irreversible reaction q = 0 also.
In a reversible reaction involving an ideal gas, ΔE still will equal zero, however, the pressure will be changing
continuously and work (w) is a function of P, work done must be determined over the entire course of the reaction. This
result in the following mathematical reduction:
w = RTln[V2/V1]

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Since in this situation ΔE = 0, q = w. This demonstrates that some of the heat of the surroundings has to be absorbed by
the system in order to perform the work of changing the system volume.
Reversible reactions differ from irreversible in that the former always proceeds infinitely slowly through a series of
intermediate steps in which the system is always in the equilibrium state. Whereas, in the irreversible reaction no
equilibrium states are encountered. Irreversible reactions are also spontaneous or favorable processes. Thermodynamic
calculations do not give information as to the rates of reaction only whether they are favorable or not.

Second Law of Thermodynamics: Entropy

The second law of thermodynamics states that the universe (i.e. all systems) tends to the greatest degree of
randomization. In other words, all naturally occurring processes proceed in a direction that leads to a minimum potential
energy level, that is, toward equilibrium. Such spontaneous reactions (as they are called) release energy as they
progress toward equilibrium and, theoretically, the energy can be harnessed and made to do work. (Recall spontaneous
reactions; heat flow from warm body into a cooler body and never in the opposite direction, water flows downhill, gases
diffuse from region of high pressure and concentration to a region of lower pressure and concentration. In all these
spontaneous reactions, energy is conserved. For example, the heat lost by the warm body is gained by the cooler body.
But certainly, something has been lost. That something is the capacity or potential to do more work ie to transfer still
more energy). While the total energy of a system and its surroundings may remain constant, the energy is distributed in
a qualitatively different way after a spontaneous reaction.

This concept is defined by the term entropy, S.

S = klnW
where k = Boltzmann constant (the gas constant, R, divided by Avagadros' number) and W = the number of substrates.
For an isothermal reversible reaction the change in entropy can be reduced to the term:
ΔS = ΔH/T
Whereas, enthalpy is a term whose value is largely dependent upon electronic internal energies, entropy values are
dependent upon translational, vibrational and rotational internal energies. Entropy also differs from enthalpy in that the
values of enthalpy that indicate favored reactions are negative and the values of entropy are positive. Together the
terms enthalpy and entropy demonstrate that a system tends toward the highest entropy and the lowest enthalpy. In
order to effectively evaluate the course (spontaneity or lack there of) of a reaction and taking into account both the first
and second laws of thermodynamics, Josiah Gibbs defined the term, free energy which is defined as:
ΔG = ΔH – TΔS
Free energy is a valuable concept because it allows one to determine whether a reaction will proceed and allows one to
calculate the equilibrium constant of the reaction which defines the extent to which a reaction can proceed. The
discussion above indicated that a decrease in energy, a negative ΔH, and an increase in entropy, a positive ΔS, are
indicative of favorable reactions. These terms would, therefore, make ΔG a negative value. Reactions with negative ΔG
values are termed exergonic and those with positive ΔG values endergonic. However, when a system is at
equilibrium:
ΔG = 0
Gibbs' free energy calculations allows one to determine whether a given reaction will be thermodynamically favorable.
The sign of ΔG states that a reaction as written or its reverse process is the favorable step. If ΔG is negative then the
forward reaction is favored and visa versa for ΔG values that are calculated to be positive.

Standard State Conditions in Biological Reactions

To effectively interpret the course of a reaction in the presence of a mixture of components, such as in the cell, one
needs to account for the free energies of the contributing components. This is accomplished by calculating total free
energy which is comprised of the individual free energies. In order to carry out these calculations one needs to have a
reference state from which to calculate free energies. This reference state, termed the standard state, is chosen to be
the condition where each component in a reaction is at 1M. Standard state free energies are given the symbol: G°.
The partial molar free energy of any component of the reaction is related to the standard free energy by the following:
G = G° + RTln[X]
From this equation one can see that when the component X, or any other component, is at 1M the ln[1] term will
become zero and:
G = G°

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The utility of free energy calculations can be demonstrated in a consideration of the diffusion of a substance across a
membrane. The calculation needs to take into account the changes in the concentration of the substance on either side
of the membrane. This means that there will be a ΔG term for both chambers and, therefore, the total free energy
change is the sum of the ΔG values for each chamber:

This last equation indicates that if [A]2 is less than [A]1 the value of ΔG will be negative and transfer from region 1 to 2 is
favored. Conversely if [A]2 is greater than [A]1 ΔG will be positive and transfer from region 1 to 2 is not favorable, the
reverse direction will be.
One can expand upon this theme when dealing with chemical reactions. It is apparent from the derivation of ΔG values
for a given reaction that one can utilize this value to determine the equilibrium constant, Keq. As for the example above
dealing with transport across a membrane, calculation of the total free energy of a reaction includes the free energies of
the reactants and products:
ΔG = G(products) - G(reactants)
Since this calculation involves partial molar free energies the ΔG° terms of all the reactants and products are included.
The end result of the reduction of all the terms in the equation is:

When the above equation is used for a reaction that is at equilibrium the concentration values of A, B, C and D will all be
equilibrium concentrations and, therefore, will be equal to Keq. Also, when at equilibrium ΔG = 0 and therefore:
0 = ΔG° + RTlnKeq

This demonstrates the relationship between the free energy values and the equilibrium constants for any reaction.

Coupled Reactions
Two or more reactions in a cell sometimes can be coupled so that thermodynamically unfavorable reactions and
favorable reactions are combined to drive the overall process in the favorable direction. In this circumstance the overall
free energy is the sum of individual free energies of each reaction. This process of coupling reactions is carried out at all
levels within cells. The predominant form of coupling is the use of compounds with high energy to drive unfavorable
reactions.
The predominant form of high energy compounds in the cell are those which contain phosphate. Hydrolysis of the
phosphate group can yield free energies in the range of –10 to –62 kJ/mol. These molecules contain energy in the
phosphate bonds due to:
1. Resonance stabilization of the phosphate products
2. Increased hydration of the products
3. Electrostatic repulsion of the products
4. Resonance stabilization of products
5. Proton release in buffered solutions
The latter phenomenon indicates that the pH of the solution a reaction is performed in will influence the equilibrium of
the reaction. To account for the fact that all cellular reactions take place in an aqueous environment and that the [H 2O]
and [H+] are essentially constant these terms in the free energy calculation have been incorporated into a free energy
term identified as:

Incorporation of the last equation into a free energy calculation for any reaction in the cell yields:

Oxidation-Reduction (Redox) Reaction

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In living systems both energy capture and energy-releasing process consist essentially or redox reaction. Redox
reaction occur when electrons are transferred between an electron donor which is the reducing agent and an electron
acceptor (oxidizing agent). In some reactions only electrons are transferred e.g
Cu+ + Fe3+ ↔ Cu2+ + Fe2+

In this reaction, Cu+ is the reducing agent, having donated an electron which was accepted by the oxidizing agent, Fe 3+.
In many reactions both electrons and protons (H+) are transferred e.g the reaction catalyzed by lactate dehydrogenase,
2 electrons and 2 protons are transferred from NADH + H to reduced pyruvate to lactate.

CH3COCOOH + NADH + H+ ↔ CH3COHCOOH + NAD+

Half Reaction and Redox Potential

Redox reactions are more readily understood when separated into half reactions e.g in the reduction of Fe 3+ by Cu+ the
Cu+ lost one electron to become Cu2+. Thus one of the half reactions in this process
Cu+ - e- ↔ Cu2+ OR Cu+ ↔ Cu2+ + e-
This equation shown clearly that Cu is the electron donor (reducing agent) and the pair of Cu + and Cu2+ constitutes
+

conjugate redox pair. The electron lost by Cu+ is accepted by Fe3+.

The separation of the redox reaction emphasizes that electrons are always the common intermediate between
half reactions. The constituent of half reactions may be observed in a special equipment called electrochemical cells.
The movement of electron generated in the half cell undergoing oxidation (Cu+ ↔ Cu2+) produces a voltage or potential
difference between the two half cells. Thus voltage is measured by a voltmeter and its sign may be +ve or –ve
depending on the direction of electrons flow. The magnitude of the p.d.f. (in volt) is a measure of the energy that drives
the reaction.
Energy Rich Compounds
The essential role of molecules such as ATP as the driving force for a large number of endergonic reactions has
led to the classification of biochemical substances as high (rich) energy or low (poor) energy compounds. Molecules like
ATP are characterized by a large negative free energy (-∆G) of hydrolysis under physiological conditions and thus
considered as energy rich, while ordinary phosphate esters are considered energy poor. An energy rich compound is
one whose reaction with a substance commonly present in the environment is accompanied by a large negative free
energy change at physiological pH. Energy rich compounds include phosphoric anhydrides (ATP, UTP, GTP, CTP etc),
acyl phosphates (acetyl phosphate, 1,3-bisphosphoglycerate), enol phosphates (phosphoenol pyruvate) and
phosphoguanidines (phosphocreatinine, phosphoarginine)
There is no clear line of demarcation between energy rich and energy poor classes but as approximate guide,
compounds exhibiting a free energy of hydrolysis at pH 7 more than -7kcal/mole and compounds exhibiting O-R
potential at pH more negative than that of O2 electrode by 0.5V may be considered energy rich. Bonds whose hydrolysis
proceeds with large negative values of more than -25kJ/mol are also energy rich compounds.