M.R.W.

-- Thermodynamics 1

1st Year Thermodynamic Lectures

Dr Mark R. Wormald

Basic chemical thermodynamics :-

Lecture 1. Introduction. Basic Definitions. 1st Law of thermodynamics.

Lecture 2. 2nd Law and entropy. Free energy and equilibria. Single component systems.

Lecture 3. Chemical potential and multiple component systems.

Lecture 4. Determining thermodynamic functions. Multiple step processes.

BIBLIOGRAPHY

Principles and Problems in Physical Chemistry for Biochemists, 3rd edition, N.C. Price, R.A. Dwek,
R.G. Ratcliffe and M.R. Wormald, Oxford University Press -- the standard ‘Oxford 1st year
biochemists’ text.

Basic Chemical Thermodynamics, E.B. Smith, Oxford Chemistry Series, Oxford University Press --
the standard ‘Oxford 1st year chemists’ text on thermodynamics -- slightly more advanced and very
good.

Physical Chemsitry for the Biomedical Sciences, S.R. Logan, Taylor & Francis.

Physical Chemistry for the Life Sciences, P.W. Atkins and J. de Paula, Oxford University Press --
more simple version of the ubiquitous ‘Physical Chemistry’ -- highly recommended.

Bioenergetics at a glance, D.A. Harris, Blackwell -- 1st few chapters give a good overview.

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 M.R.W. -- Thermodynamics 1

CONSTRAINTS IMPOSED ON BIOLOGICAL SYSTEMS BY PHYSICS AND

CHEMISTRY

In order to exist, a living species has to:

1. collect energy from the surroundings
2. convert it to a useful form (ion gradients, ATP, etc)
3. use it to grow, change, replicate, etc.

Living systems can only do something if the energy needed is balanced by the energy that is available.

THERMODYNAMICS
“The Study of Heat and Work”

Thermodynamics allows us to investigate the distribution of energy in a system.

By comparing two systems, we can determine how much energy is needed to convert, or given out by
converting, one systems to another.

We can apply thermodynamics to stable systems, unstable systems, or to each stage of a reaction
mechanism.

Thermodynamics cannot give information on reaction rates directly (although the amount of energy
required is going to be important in determining reaction rates - see Transition State Theory).

FEATURES

1. It is an empirical model -- we use it because it works.

2. It is based on a few initial postulates, the Laws of Thermodynamics, everything else follows
logically.
3. It makes no assumptions about the nature of a system -- we do not even have to know whether or
not molecules exist.

STANDARD STATES/CONDITIONS

As we are mostly comparing systems, it is useful to have constant reference points (standard states)
against which all comparisons can be made.

Standard states for compounds are defined as :

Solid pure solid

Liquid pure liquid
Solute concentration of 1 mol per kg of solvent in a given solvent
Gas pure gas at 1 atm pressure

Standard conditions for all reactions of practical interest are defined as 1 atm pressure and 298 K.

Biochemical standard conditions are those in which all components are present in their standard
states, except H+ which is present at 10-7 mol dm-3, i.e. pH = 7.0.

TYPES OF SYSTEMS

Open -- systems that can exchange matter and energy with the surroundings

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 M.R.W. -- Thermodynamics 1

Closed -- systems that can only exchange energy with the surroundings. Almost all chemical reactions
fall into this category.

Isolated -- systems that cannot exchange either energy or matter with the surroundings. The most
obvious case of an isolate system is the universe.

STATE FUNCTIONS

A state function depends only on the state of the system, not on the path taken to reach that state.

Energy (E)
State B
∆E(AB)

∆E(B)°

State A

∆E(A)°

Standard State

Intensive -- the property is independent of the size of the system (it has the same value for a small
region of the system as for the whole system). e.g. temperature, pressure.

Extensive -- the property is dependent on the size of the system (the value changes between a small
region of the system and the whole system). e.g. mass, volume, energy.

Most thermodynamic quantities are extensive state functions.

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 M.R.W. -- Thermodynamics 1

1st LAW OF THERMODYNAMICS

(The Law of Conservation of Energy)

The algebraic sum of all energy changes in an isolated system is zero.

We define U as the internal energy of a system. From the 1st Law, U must be a state function. Thus;

∆U = Ufinal - Uinitial

For an isolated system, from the 1st Law ∆U = 0.

For a closed system, the internal energy of the system can increase as long as the internal energy of
the surroundings decrease by the same amount.

∆Usystem = - ∆Usurroundings

Work and heat :-

As we shall see, it is useful to distinguish between two classes of energy, work and heat. Again, we
can define;

q - heat absorbed by the system from the surroundings

w - work done on the system by the surroundings

∆Usystem = q + w

Distinction between work and heat :-

Work is the energy associated with orderly movements of bodies, for example pushing back
boundaries (volume change) or moving electrons along a wire.

Heat is the energy associated with disorderly movements of bodies, for example random molecular
motion in a gas.

Work in chemical systems :-

The work energy term (w) is also the sum of many different types of energy, the most common being
electrical work (such as electrochemical cells), mechanical work (such as muscle contraction) and
change of volume (expansion).

w = welectrical + wmechanical + wexpansion

Most of these are usually zero. However, at constant pressure (where life exists) the volume of the
system often changes and thus does work by pushing back the surroundings. In this case, w is given
by

wexpansion = - P∆V

∆V is the increase in volume of the system. w is negative because, by increasing in volume, the system
is doing work on the surroundings.

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 M.R.W. -- Thermodynamics 1

ENTHALPY

It is usually convenient to factor out the changes in internal energy due to changes in volume (ie.
allow for constant pressure). Thus;

Change in internal energy allowing for volume = ∆U - wexpansion

changes

= ∆U + P∆V

= (Ufinal + PVfinal) - (Uinitial + PVinitial)

(U + PV) is an extensive state function and is called, for convenience, ENTHALPY, H.

H = U + PV

∆H is a measure of the change in the total energy of a system after allowing for changes in volume.

For most chemical systems, the only other form of energy that is exchanged with the surroundings is
heat. ∆H is then simply the heat absorbed at constant pressure.

DIRECTION OF A CHEMICAL REACTION

Dissolution of NaNO3 Spontaneous Endothermic (∆H = +ve)

Dissolution of NaOH Spontaneous Exothermic (∆H = -ve)

Diffusion of Na+ in water Spontaneous ∆H = 0

Thus, consideration of enthalpy (or internal energy) does not give us any information about the
direction of a chemical reaction. In fact, in any isolated system (such as the universe) ∆H = 0 from the
1st law, yet changes do occur in the universe.

The 1st law of thermodynamics can be thought of as an accounting tool, used to keep track of energy
during a reaction.

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 M.R.W. -- Thermodynamics 2

CHANGE AND WORK

Change, such as a chemical reaction, is an orderly process.

Work (w) ⇒ energy associated with an orderly process
Heat (q) ⇒ energy associated with a disorderly process
Thus, when considering change we need to consider work energy not total energy.

For a change to occur, we need to either;

1. Put work energy into the system (drive change externally).
or;
2. Use some of the work energy already present in the system (spontaneous change).
If there is not enough available work energy, no change can occur.

2nd LAW OF THERMODYNAMICS

Spontaneous changes are those which, if carried out under the proper conditions, can
be made to do work. If carried out reversibly they yield a maximum amount of work.
In irreversible (spontaneous) processes the maximum work is never achieved.

ENTROPY

We need a measure of the available work energy in a system. We do this by determining the unavailable
(non work) energy in a system. This is just the heat energy, q.

We define the ENTROPY, S, of a system as

δq reversible final dq reversible

δS = or ∆S = ∫initial
T T

S is a measure of the randomness of the system. T∆S is a measure of the change in the
heat energy of the system (energy unavailable to do any further work).

For any system;

Change in total energy = Change in work energy + Change in heat energy

∆H - change in total energy (after allowing for expansion)

T∆S - change in heat energy

Thus;
∆H = Change in work energy + T∆S
or;
Change in work energy = ∆H - T∆S

If a change has occurred, then we must have used work energy to make the change happen and so the
change in work energy must be -ve.

Isolated system :-

There is no external source of energy. Only internal work energy can be used.

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 M.R.W. -- Thermodynamics 2

∆H = 0 ⇒ Change in work energy = -T∆S

The amount of work energy can never increase. Thus;

∆S ≥ 0

If ∆S = 0, there is no work energy being used and so the system does not change (equilibrium).
If ∆S > 0, work energy is being used and so the system can change.

Closed systems :-

External energy is available.

∆H ≠ 0 ⇒ Change in work energy = ∆H-T∆S

The amount of work energy can never increase during a change. Thus ∆S ≥ ∆H/T.

GIBBS FREE ENERGY

Change in work energy = ∆H - T∆S

This can be rewritten as;

Change in work energy = (Hfinal - TSfinal) - (Hinitial - TSinitial)

(H - TS) is an extensive state function and for convenience we call it the GIBBS
FREE ENERGY, G.

Change in work energy = ∆G

At equilibrium ∆G = 0. For an irreversible (spontaneous) reaction ∆G < 0. This gives us a general

criteria for whether a reaction will occur or not in a closed or isolated system.

POSITION OF EQUILIBRIUM

∆G can be used to determine how far a reaction will go by letting the system change a small amount and
then recalculating ∆G.

End point of
1 the reaction

∆G < 0

∆G > 0
∆G = 0

Reactants Products
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 M.R.W. -- Thermodynamics 2

CONTROL OF REACTIONS

Kinetic control - 1
The products are not in equilibrium with the reactants. [This does not mean that an equilibrium cannot
be established, only that is has not been established so far].

The amount of product depends on the amount of reactant, the speed of the reaction and how long it has
been going.

The reaction cannot be made to go backwards by increasing the concentration of the products (until
after equilibrium has been established).

Thermodynamic control - 2
The products are in equilibrium with the reactants.

The ratio of products to reactants depends on the relative energies of the two states (see later lectures).

The reaction can be made to go forwards or backwards by increasing the concentrations of the reactants
or products respectively.

SINGLE COMPONENT SYSTEMS

A system consisting of one chemical can exist in three states, solid liquid and gas.

H - total energy in the system is related to the energy involved in the bonds between the molecules;
Solid - H small and positive
Liquid - H medium and positive
Gas - H large and positive (lots of energy necessary to overcome bonds)

S - the randomness of the system depends on how free the molecules are to move around relative to
each other;
Solid - S small
Liquid - S medium
Gas - S large

PHASE CHANGES

A plot of G versus temperature for any given phase will be a straight line with a slope of -S (G = H -
TS). At any given temperature, the most stable phase will be the one with the lowest value of G.

Solid

Melting
Liquid
point Boiling
point Gas
T
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 M.R.W. -- Thermodynamics 2

EFFECTS OF PRESSURE ON PHASE EQUILIBRIA

Solid and liquid - virtually incompressible, thus pressure has no effect.

Gas - increasing the pressure will effect the properties of the molecules (they will collide more often).
By compressing the gas, work is being done on the system and so G goes up. Note - it does not matter
how you increase the pressure (e.g. compressing the system, adding another inert gas, etc.)

Solid

Melting
point Liquid
Gas (low P) Gas (high P)

T
For an ideal gas -

G = G° + RT In (P/P°)n

Free energy of the Free energy of the Variation in free energy of the
compound compound in the standard compound with composition -
state - depends on its depends on chemical environment
chemical properties

PHASE DIAGRAM FOR WATER

Plot of the most stable phase at a given temperature and pressure.

Solid-liquid phase change - independent of pressure (to a first approximation).
Liquid-gas phase change - occurs at a lower temperature as the pressure is reduced.
Below a given pressure, the liquid phase will not be stable and the solid will convert directly to the gas.

P
Liquid
Solid

1 atm
Gas

T
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 M.R.W. -- Thermodynamics 2

CONCENTRATION/DRYING OF PROTEINS

1. Increase the temperature so the liquid boils - the heat denatures the protein.

2. Reduce the pressure so that the liquid boils - this causes bubbling which denatures the protein.

3. Just leave to evaporate - at very low water content, the protein structure changes to compensate for
the lack of solvent (denatures). This is also a problem with 1 and 2.

4. Freeze the sample to form a solid (usually at about 80 K) and then reduce the pressure so that the
solid changes directly to a gas (called lyophilisation) - leaves the protein structure intact.

This technique is also used for;

• Drying/concentrating other biological samples (e.g. DNA)

• Drying foods for later rehydration (coffee, astronauts rations)
• Recovering paper after water damage

PROTEIN FOLDING / UNFOLDING

Folded protein :-

H - small and positive

S - small and positive

Unfolded protein :-

H- large and positive

S - large and positive

Folded

Unfolded

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 M.R.W. -- Thermodynamics 2

EFFECT OF PRESSURE ON G FOR A GAS

- derivation

G = H - TS : H = U + PV

Thus;
G = U + PV - TS

A very small change in G is given by;

δG = δU + PδV + VδP - TδS - SδT

For a closed system, from the 1st Law;

δU = δq - PδV

At equilibrium, from the 2nd Law;

δq = TδS ⇒ δU = TδS - PδV

Combining all of these gives;

δG = VδP - SδT

At constant temperature, δT = 0. Thus;

δG = VδP

For a perfect gas PV=nRT. Thus;

δG = nRT/P δP

Integrating this gives;

final final 1
∫initial dG = nRT ∫initial P dP

Thus;
n
 P final 
G final - G initial = RT In 
 Pinitial 

 P final 
Note that   is a dimensionless quantity, otherwise we could not take logs of it.
 Pinitial 

We normally take the initial state to be the standard state of the gas. Thus;

Ginitial = G° : Pinitial = P° = 1 atm

Thus;

n
 P 
G = G o + RT In o 
P 

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 M.R.W. -- Thermodynamics 3

CHEMICAL REACTIONS

Thermodynamics is only interested in comparing the state of a system before and after a change has
occurred (and is not interested in the mechanism of that change).

A chemical reaction, in thermodynamic terms, simply consists of taking away reactants and adding
products.

Reactants Products

∆G = ∆G (removing reactants) + ∆G (adding products)

CHEMICAL POTENTIAL

The chemical potential of a species is defined as the change in free energy (G) of a system when 1 mole
of the species is added to the system at constant temperature and pressure†. It is an intensive state
function.

System + 1 mole of A
System + A

∆G = Chemical potential of A (µA)

If instead of adding 1 mole of A we add δnA moles of A, then

∆G = µA . δnA

For any chemical reaction, ∆G (at constant temperature and pressure) is simply given by adding up the
changes in each component.

∆G = Σi µi δni

† Note - more formally, µ is the rate of change of G with composition.

 dG 
µi =  
 dn i  T,P,n
j

EQUILIBRIUM AND CHEMICAL POTENTIAL

Consider a system consisting of a component distributed between two phases, a and b. The chemical
potentials in the two phases are µa and µb. At constant temperature and pressure, if we transfer δn moles
from a to b then;

dn
µa µb

∆Ga = - µa.δn -- (negative because matter is being lost)

∆Gb = + µb.δn -- (positive because matter is being gained)

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 M.R.W. -- Thermodynamics 3

Overall;
∆G = ∆Gb + ∆Ga = µb.δn - µa.δn

∆G = (µb - µa).δn

At equilibrium, ∆G = 0. Thus;
µb = µa.

Equalising the chemical potentials is the driving force for any reaction.

VARIATION IN µ WITH COMPOSITION

For a pure substance

µ=G and µº = Gº

by definition. Thus, for a pure gas (by analogy with the equation for G);

µ = µ° + RT ln (P/P°)

Unlike G, we can also use µ to describe a single component in a mixture. For one species (i) in a
mixture of perfect gases;
µi = µi° + RT ln (Pi/P°)

where Pi is the partial pressure of i.

We can derive a very similar equation for liquids and solutions;

µx = µx° + RT ln ([x]/[x]°)

where [x] is the concentration of x in the liquid and [x]° is its concentration in its standard state.

For solids, G does not change and so neither does µ (µi = µi°).

FREE ENERGY OF A REACTION

∆G for a chemical reaction at constant pressure and temperature is given by;

∆G = Σi µi δni

For each component;

µx = µx° + RT ln ([x]/[x]°)
Thus;
 [x] 
∆G = ∑ µox δnx + ∑ RTδnx ln  o 
x x  [x] 

  [x] δnx 
= ∑ x x
µ o
δn + RT ln ∏ o 
 x  [x] 


x
 

δnx is negative for reactants and positive for products.

Σx µx° δnx is the standard free energy change for a mole of reactants giving a mole of products, ∆G°.

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 M.R.W. -- Thermodynamics 3

δnx for a complete reaction is the stoichiometry coefficient (mx) for that component.

The [x]° terms are often left out because they usually equal 1.0 (although this can lead to confusion).

Thus;

  [x] 
mx

 ∏ products  o  
  [x]  
∆G = ∆Go + RT ln  mx 
∏  [x]  
 reactants  [x]o  
   

EQUILIBRIUM CONSTANTS

At equilibrium, the ratio of products to reactants is called the equilibrium constant, K.

  [x] 
mx

 ∏ products  o  
  [x] eq 
K=  mx 
  [x]  
 ∏ reactants  [x]o  eq 
 

K is a dimensionless quantity (it has no units), but [x] must have the same units as [x]°.

At equilibrium ∆G = 0, by definition. Thus;

∆G° = -RT ln (K)

or in the biological standard state;

∆G°′ = -RT ln (K′)

Note - ∆G° and ∆G°′ (and thus K and K′) are only different if H+ is involved in the reaction because x°
is different for H+.

HYDROLYSIS OF ATP

Consider the hydrolysis of ATP to ADP and inorganic phosphate;

ATP (aq) + H2O ↔ADP (aq) + Pi (aq) + H+ (aq)

Measuring concentrations of an equilibrium mixture of ATP, ADP and Pi at pH = 7 and T = 310K gives
K’ = 1.3 ×105 (M). Thus;

[ADP]eq [Pi ]eq [H+ ]eq [ADP]eq [Pi ]eq  - ∆Go' 

K' = = = exp  
[ATP]eq [H2 O]eq [ATP]eq  RT 

and so ∆Gº' = -30.5 kJ mol-1.

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 M.R.W. -- Thermodynamics 3

Note: [H+] = [H2O] = 1.0 because they are in their standard states (pH = 7.0 for the former and
approximately a pure liquid for the latter). If we were considering K rather than K′, then [H+] would
equal 10-7 at pH=7.0.

Hydrolysis of ATP to ADP in cells :-

The concentrations of ATP, ADP and Pi in a cell are found to be;

[ATP] = 1 x 10-2 mol.dm-3

[ADP] = 3 x 10-3 mol.dm-3
[Pi] = 1 x 10-3 mol.dm-3

If the system were at equilibrium;

[ADP]eq [Pi ]eq
[ATP] eq = '
= 2.3 x 10 -11 mol dm -3
K
Thus, the cell is not at equilibrium. If it were at equilibrium, no work could be obtained from the
hydrolysis of ATP (∆G = 0).

∆G in the cell is given by;

 [ADP][Pi ]
∆G = ∆G o' + RT ln  = - 51.4 kJ mol
-1
 [ATP] 

∆G is large and negative, thus considerable work can be done in the cell by hydrolysing ATP.

MASS ACTION RATIO (MAR)

The mass action ratio is defined as;

∏ products [x] m
MAR =
∏ reactants [x] m
Thus;
∆G = ∆G° + RT ln (MAR)

If the system is at equilibrium,

MAR = K

If the system is not at equilibrium then the difference between the MAR and K gives a measure of how
far from equilibrium the system is.

∆G = RT ln(MAR/K)

As a rough rule of thumb, if MAR is more than 2 orders of magnitude (100 times) smaller than K in a
biological system, then that reaction is likely to be kinetically controlled rather than thermodynamically
controlled.

HYDROLYSIS OF ATP -- again

ATP (aq) + H2O ↔ADP (aq) + Pi (aq) + H+ (aq)

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 M.R.W. -- Thermodynamics 3

For this reaction;

'
[ADP] eq [ Pi ]eq [H + ] eq
K = = 1.3 x 10 5
[ATP] eq [H 2 O] eq

For the observed concentrations of ATP, ADP and Pi in a cell;

[ATP] = 1 x 10-2 mol.dm-3

[ADP] = 3 x 10-3 mol.dm-3
[Pi] = 1 x 10-3 mol.dm-3

Thus;
[ADP] [ Pi ] [H + ]
MAR = = 3 x 10 -4
[ATP] [H 2 O]

High-energy phosphate bonds;

Because hydrolysis of ATP involves breaking a phosphate bond and the equilibrium lies along way to
the right, this is called a high-energy phosphate bond, and is often interpreted as the phosphate bond
being weak.

It requires a great deal of energy to break a bond (for O-P ≈ 540 kJ mol-1). If energy was given out
when a bond is broken, then the bond would never be formed in the first place.

Thermodynamics (in this case K and ∆G) does not give us any information about a small part of the
system, only about the difference between the state of the whole system before and after the reaction.

∆G = Gfinal - Ginitial = ∆H - T∆S

In this case;

∆H - depends on the stability of the reactants and the products, one component of which is the
energy of the bonds that need to be broken or formed (including with the solvent).

∆S - depends on the number of reactant and product molecules and on the ordering effect they
have on the solvent.

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 M.R.W. -- Thermodynamics 4

MEASUREMENT OF THERMODYNAMIC QUANTITIES

K Measure concentrations at equilibrium.

∆Gº Calculate from ∆Gº = -RT ln(K).
∆G Measure concentrations and calculate from ∆G = ∆Gº + RT ln(MAR).
∆Hº Measure temperature dependence of K.
∆H Measure directly by calorimetry.
∆Sº Calculate from ∆Gº = ∆Hº - T∆Sº
∆S Calculate from ∆G = ∆H - T∆S

∆Gº, ∆Hº, ∆Sº can also be calculated via Hess’s Law from standard values.

VARIATION OF EQUILIBRIUM CONSTANT WITH TEMPERATURE

As ∆Gº = - RT ln(K), then;

∆Go ∆Ho ∆So
ln K = − = − +
RT RT R
Assuming that ∆Hº and ∆Sº are independent of temperature and differentiating with respect to T gives;
d ln K ∆Ho
=
dT RT 2
This equation is called the Van't Hoff Isochore. Reintegrating this equation gives;
K  ∆Ho  1 1 
ln  2  = −  − 
 K1  R  T2 T1 
Thus, a plot of ln(K) vs. 1/T should give a straight line of slope -∆Hº/R.

MULTIPLE STEP REACTIONS

Many reactions involve several discrete steps.

Reactants ↔ Intermediates ↔ Products

In this case, we can apply our standard thermodynamic analysis to each step in turn.

Reactants ↔ Intermediates ....(1)

∏ [intermediates]n 
∆G(1) = ∆G(1) o + RT ln  intermediates 

 ∏reactants [reactants] n


and

Intermediates ↔ Products ....(2)

∆G(2) = ∆G(2) + RT ln
o

 ∏ products
[products]n 

∏ n
 intermediates [intermediates] 

However, since G is a state function (it only depends on how the system is at the start and at the end),
we must also be able to apply our standard equations to the whole reaction regardless of what the
intermediates are.

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 M.R.W. -- Thermodynamics 4

Reactants ↔ {Intermediates} ↔ Products ....(3)

 ∏products [products]n 
∆G(3) = ∆G(3) + RT ln 
o 
∏ [reactants] n
 reactants 

From this it also follows that;

∆G(3) = ∆G(1) + ∆G(2) and ∆G(3)° = ∆G(1)° + ∆G(2)°

COUPLED REACTIONS

Sequential reactions :-
A ↔ B ↔ C : ∆G (AB) > 0, ∆G(BC) << 0

∆G (AC) = ∆G (AB) + ∆G (BC) < 0

Thus, A will still be converted into C although the conversion from A to B is unfavourable.

Parallel reactions :-
A ↔ B : ∆G (AB) > 0
C ↔ D : ∆G (CD) << 0

The first reaction would normally not proceed, the second would proceed. If the two reactions are
coupled, then;

A+C ↔ B+D : ∆G = ∆G (AB) + ∆G (CD) < 0

and the reaction will proceed converting A into B.

GLUTAMINE SYNTHESIS

Glutamine, used both as an amino-acid and a non-toxic source of ammonia for other biochemical
pathways, is produced in cells from glutamate.

Glutamate(aq) + NH3(aq) ↔ Glutamine(aq) + H2O ∆G°‘ = + 14.3 kJ mol-1

This reaction would not normally proceed. However, the free energy necessary to drive the reaction
forward 14.3 kJ mol-1 is less than the free energy given out by hydrolysis of ATP.

ATP(aq) + H2O ↔ ADP(aq) + Pi(aq) ∆G°‘ = - 30.5 kJ mol-1

Thus, the coupled reaction;

ATP(aq) + Glutamate(aq) + NH3(aq) ↔ ADP(aq) + Pi(aq) + Glutamine(aq)

∆G°‘ = + 14.3 - 30.5 = - 16.2 kJ mol-1

will proceed spontaneously. These two reactions are coupled together by the enzyme glutamine
synthetase

THE TRIOSE PHOSPHATE ISOMERASE REACTION

The reaction

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 M.R.W. -- Thermodynamics 4

fructose dihydroxyacetone glyceraldehyde

ƒ +
1,6-bisphosphate phosphate 3-phosphate
-3
has an equilibrium constant of 1.5×10 and is catalysed by the enzyme aldolase. Thus, it appears that
this equilibrium lies to the left.

The next step in the glycolytic pathway uses G-3-P and so dha-P needs to be converted to G-3-P. The
reaction
dihydroxyacetone glyceraldehyde
ƒ
phosphate 3-phosphate
has an equilibrium constant of 0.04 and is catalysed by the enzyme triose phosphate isomerase. This
equilibrium also appears to lie to the left.

Aldolase is added to a 4 mM solution of fructose 1,6-bisphosphate. At equilibrium, assume x mM of G-

3-P has been produced, so the equilibrium concentrations are:

[F-1,6-P] = 4-x mM : [G-3-P] = [dha-P] = x mM

Thus
[dha-P][G-3-P] (x ×10-3 )2
K= = = 1.5 ×10−3
[F-1,6-P] (4 − x) ×10-3

This gives
[F-1,6-P] = 2.23 mM : [G-3-P] = [dha-P] = 1.77 mM

Even though the equilibrium constant is small (1.5×10-3), at these concentrations the reaction proceeds
to approx. 50%.

Aldolase and triose phosphate isomerase are added to a 4 mM solution of fructose 1,6-bisphosphate. At
equilibrium, assume x mM of F-1,6-P has been used and y mM of G-3-P has been converted to dha-P,
so the equilibrium concentrations are:

[F-1,6-P] = 4-x mM : [G-3-P] = x–y mM : [dha-P] = x+Y mM

Thus;
(x − y) × 10−3 × (x + y) × 10−3 −3 (x − y) × 10−3
= 1.5 × 10 and = 0.04
(4 − x) ×10−3 (x + y) × 10−3

This gives
[F-1,6-P] = 0.93 mM : [G-3-P] = 0.24 mM : [dha-P] = 5.90 mM

Addition of triose phosphate isomerase to the products of the aldolase reaction lowers the equilibrium
concentration of G-3-P by nearly a factor of 10.

At first sight this is unfavourable, as G-3-P is the reactant for the next step of the glycolytic pathway.
However, this subsequent reaction will reduce the concentration of G-3-P (i.e. is sequentially coupled to
the aldolase and triose phosphate isomerase reactions), which will result in more dha-P being converted
to G-3-P. In the context of the whole pathway triose phosphate isomerase increases the thermodynamic
efficiency by enabling dha-P to be used.

BIOSYNTHETIC PATHWAYS

A biosynthetic pathway consists of a series of sequentially coupled reactions. This enables the free
energy from one reaction to be used by another reaction.
• Reactions that are only slightly unfavourable thermodynamically are driven by
sequential coupling in the pathway.

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 M.R.W. -- Thermodynamics 4

• Reactions that have very low equilibrium constants are coupled directly (in parallel)
to energy producing reactions, such as ATP hydrolysis.
One of the evolutionary selection pressures is the availability of energy, and so pathways need to be
reasonably efficient from a thermodynamic point of view.

∆G°’ FOR GLYCOLYSIS

The net reaction for glycolysis is

ˆˆ† 2 pyruvate + 2 NADH + 2 ATP + 2 H + + 2 H 2 O

glucose + 2 NAD + + 2 ADP + 2 Pi ‡ˆˆ

The free energies of formation of the reactants and products are

∆G°’(formation) (kJ mol-1) ∆G°’(formation) (kJ mol-1)

ADP -1409.00 NAD+ 1059.11
ATP -2276.77 NADH 1120.09
Glucose -426.71 Pi -1059.49
H+ 0.00 Pyruvate -350.78
H2 O -155.66

Thus
∆G°’ = ∆G°’f(products) - ∆G°’f(reactants) = -80.77 kJ mol-1

∆G FOR GLYCOLYSIS

The metabolite concentrations measured in erythrocytes are as follows:

Concentration (mM) Concentration (mM)

ADP 0.14 Pi 1.00
ATP 1.85 Pyruvate 0.05
Glucose 5.00

Assuming that the NAD+/NADH ratio is 1 and the cytoplasmic pH is 7,

[pyruvate]2 [ATP]2 [NADH]2

∆G = ∆G o ' + RT ln = −69.6 kJ mol-1
[glu cos e][ADP]2 [Pi]2 [NAD + ]2
This corresponds to the free energy released (wasted) during the glycolytic pathway (i.e. about one
ATPs worth over 10 coupled reactions). Infact, this energy is not all wasted because it is necessary to
make the pathway (i) irreversible (otherwise a small decrease in glucose concentration would lead to
glucose synthesis) and (ii) proceed at a reasonable rate.

THE GLYCOLYTIC PATHWAY

Theory of Flux Generating Steps –

Flux through (kinetics of) a biosynthetic pathway is a measure of the overall rate for conversion of the
initial reactant into the final product. This is governed by reactions that are not in equilibrium (under
kinetic control). These have two functions;
• They make the pathway irreversible (it cannot go backwards).
• They indirectly control the reactions that are in equilibrium.
Reactions that are at equilibrium play no role in controlling the flux.

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 M.R.W. -- Thermodynamics 4

Step K MAR MAR/K

Brain Heart

Hexokinase 3.09-5.50 ×103 0.04 0.08 10-5

PGI 0.36-0.47 0.22 0.24 1
PFK 0.09-1.20 ×102 0.13 0.03 10-3
Aldolase 6.08-13.0 ×10-5 2.4×10-6 9×10-6 10-1
TIM 3.06-4.50 ×10-2 - 0.24 10
GAPDH 0.02-1.50 ×103 5.3 9.0 10-2
PGM 0.01-0.02 0.2 0.12 1
Enolase 2.08-4.60 3.6 1.4 1
Pyruvate kinase 0.20-2.00 ×104 5.4 40.0 10-3

The theory of flux generating steps is flawed because it analyses reactions independently. As we have
seen, at a thermodynamic level (and a kinetic level) all reactions in a pathway are coupled and the
system needs to be analysed as a whole. This approach is called Metabolic Control Analysis.

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