Classroom session III.
Bioenergetics
Classroom practice III: Bioenergetics
BIOENERGETICS EXERCISES
Answer key provided at the end of the questionnaire
1. A solution containing 10 mM NADH and 1 mM
NAD , is mixed with the same volume of another
+
solution containing 0.01 M acetaldehyde and 0.01 M
ethanol.
a) Calculate the redox potential difference
between the reactant pairs.
b) Write the spontaneous reaction.
2. a) Write the spontaneous reaction if 1 mol of
the following compounds (in the presence of the
suitable catalyzer) is mixed: FMN, FMNH , NAD and
2
+ sugar-P + H O <-----> sugar + Pi
NADH.
b) Identify the oxidant pair.
c) Which of them is the reducing agent?
3. Given the hypothetical reaction (at 25 ºC, pH 7.0):
A<––––––>B+C
Where the initial concentration of A is 0.2 M and
having only 1% remaining when the equilibrium is
reached:
a) Calculate K’eq
b) Calculate K’eq of the reverse reaction
c) In which direction is the reaction
spontaneous?
4. Using the data from the redox potentials table,
calculate the standard free energy change of the
reduction of nitrate by NADH.
5. Calculate the apparent standard free energy change
and the apparent equilibrium constant at 25 °C and pH
7.0 for the oxidation of the reducing agent AH by the
2
oxidant compound B, knowing that:
E' (A / AH ) = -0.45 V
o 2
E' (B / BH ) = 0.25 V.
o 2
6. A solution containing 0.2 M dehydroascorbate and
0.2 M ascorbate is mixed with an equal volume of a
solution containing 0.01 M acetaldehyde and 0.01 M
ethanol at 25 °C and pH 7.0:
a) Write the thermodynamically favourable
reaction.
b) Calculate the standard redox potential
difference and the standard free energy change for the
above-mentioned reaction.
7. a) Write the spontaneous reaction that might
take place when 1 mol of each of the following
compounds is mixed: oxalacetate, malate, acetaldehyde
and acetate in the presence of the suitable enzymes to
catalyse the redox reaction between them.
b) Identify the compound that is oxidized and
the one that is reduced.
c) Calculate the standard free energy change.
d) Calculate the apparent equilibrium constant.
8. One litre of a solution containing: 10 mmol of
acetate, 5 mmol of acetaldehyde, 2 mmol of malate and
1 mmol of oxalacetate at 25 °C and pH 7.0:
a) Write the spontaneous reaction in the
presence of the suitable catalyzer.
Degree in Veterinary Medicine. Biochemistry
1
b) Identify the compound that is oxidized
c) Identify the compound that is reduced
d) Calculate the free energy change in the
above indicated conditions.
9. Write the spontaneous reaction that will take place
and calculate the free energy change when the enzyme
lactate dehydrogenase is added to a solution containing
pyruvate, lactate, NAD and NADH at 25 °C and pH
+
7.0 at the following ratios:
a) [lactate] / [pyruvate] = 1;
[NAD ] / [NADH] = 1
+
b) [lactate] / [pyruvate] = 159;
[NAD ] / [NADH] = 159.
+
c) [lactate] / [pyruvate] = 1,000;
[NAD ] / [NADH] = 1,000.
+
10. The free energy change for the hydrolysis of one
phosphoric ester from a sugar:
2
is -25.96 kJ/mol under physiological conditions within
the cells, where the steady-state concentrations of
sugar-P, sugar and Pi are 0.001 M, 0.0001 M and
0.05 M, respectively. Calculate the standard free
energy change for the reaction.
11. Calculate the free energy change for the hydrolysis
of ATP at 25 °C and pH 7.0:
ATP + H 0 <---> ADP + Pi
2
knowing that the apparent standard free energy change
is -30.5 kJ/mol, and if the ATP and ADP
concentrations were identical and the Pi concentration
was: a) 1 M; b) 0.1 M; c) 0.01 M; d) 0.001 M.
12. Calculate the free energy change for the hydrolysis
of ATP under physiological conditions, at 37 °C and
pH 7.0, knowing that the apparent standard free energy
change is -30.5 kJ/mol, and the intracellular
concentrations of ATP, ADP and Pi, in the steady-state,
are 1mM, 0.1 mM and 10 mM respectively.
13. Calculate the steady-state concentration of ATP in
the erythrocytes, knowing that ADP concentration is
0.00014 M, Pi concentration is 0.001 M and ∆G' for the
hydrolysis of ATP is -48.1 kJ/mol. (∆G'o for the
hydrolysis of ATP = -30.5 kJ/mol).
14. The apparent standard free energy change for the
hydrolysis of ATP at 25 °C and pH 7.0 is -30.5 kJ/mol.
Knowing that the apparent free energy change for the
hydrolysis of glucose-6P, under the same conditions, is
-15.88 kJ/mol, calculate:
a) ∆G'o
b) K'eq for the reaction between ATP and glucose
catalyzed by the enzyme hexokinase:
D-glucose + ATP <---> D-glucose-6-P + ADP
15. Assuming that the following redox reaction:
A + B <---> A + B
2- 2-
was coupled with the phosphorylation of ADP to give
ATP, calculate:
a) The minimal redox potential difference
needed between the two redox pairs for the reaction to
be sufficient to synthesize one molecule to ATP,
knowing that under intracellular conditions ∆G' for the
hydrolysis of ATP is -48.1 kJ/mol.
b) Repeat the same calculation assuming the
coupling efficiency is only 40%.
Classroom session III. Bioenergetics
16. In a cell, the steady-state concentration of a
phosphorylated compound, R-O-P and its hydrolysis
products, R-OH and Pi are 2 x 10 M, 4 x 10 M and
-2 -5
5 x 10 M, respectively. Under these conditions, ∆G'
-2
for the reaction:
R-O-P + H O <-----> R-OH + Pi
2
is -34.4 kJ/mol.
a) Calculate K’eq for the reaction.
b) Calculate ∆G’o for the following reaction:
R-O-P + ADP <-----> R-OH + ATP
17. Calculate ∆G’ for the hydrolysis of ATP to ADP
and Pi under the conditions in a muscle cell at rest:
[ATP] = 5 mM; [ADP] = 0.5 mM; [Pi] = 5 mM at
37°C, ∆G’o = -30.5 kJ/mol.
18. Glucose 6-P is hydrolysed enzymatically at pH 7.0
giving glucose and Pi:
D-glucose-6-P + H O <---------> D-glucose + Pi
2
knowing that the initial concentration of glucose-6-P is
0.1 M and that at the equilibrium remains only 0,05 %
of this metabolite, calculate:
a) K'eq for the hydrolysis of glucose-6-P.
b) K'eq for the synthesis of glucose-6-P from
glucose and Pi.
19. The apparent equilibrium constant for the reaction
catalyzed by the phosphoglucose isomerase is 0.43.
D-glucose-6-P <---> D-fructose-6-P
a) Calculate the standard free energy change.
b) Indicate the direction of the spontaneous reaction.
20. Knowing the standard free energy change for the
reaction catalyzed by the phosphoglucomutase:
D-glucose-6-P <--------> D-glucose-1-P
is 7.5 kJ/mol in the direction of the formation of
glucose-1-P:
Calculate final concentrations obtained after adding
appropriate amount of enzyme to a solution of 0.1 M of
glucose-1-P.
21. The enzyme glutathione reductase catalyzes the
reduction of glutathione by NADPH + H as follows:
+
GS-SG + NADPH + H <------> 2 GSH + NADP
+ +
which apparent equilibrium constant is 910, at 37°C
and pH 7.0. Calculate the apparent redox potential for
the pair GS-SG / 2 GSH.
22. The enzyme phosphoglycerate mutase catalyzes
the following reaction:
3-P-glycerate <-----> 2-P-glycerate
a) Knowing that the apparent equilibrium
constant at pH 7.0 and 37°C is 0.165, calculate the
apparent standard free energy change.
b) Knowing that within the erythrocytes the
steady-state concentrations for the two metabolites are:
[3-P-glycerate] = 61.2 µM and [2-P-
glicerate] = 4.3 µM, calculate the free energy change
for the reaction under the above conditions and predict
the effect of these concentrations on the spontaneity of
the reaction.
Degree in Veterinary Medicine. Biochemistry
2
23. The apparent equilibrium constant for the reaction
catalyzed by the phosphoglucomutase:
D-glucose-1-P <-----> D-glucose-6-P
is 19 at 37°C and pH 7.0 in the direction of the
production of glucose-6-P. Calculate the free energy
change and indicate the direction of the spontaneous
reaction with the following concentration of
metabolites:
a) [glucose-1-P] = 72 mM, [glucose-6-P] = 72 mM.
b) [glucose-1-P] = 7.2 mM, [glucose-6-P] = 137 mM.
c) [glucose-1-P] = 5.55 mM, [glucose-6-P] = 555 mM.
24. The apparent standard free energy change for the
reaction catalyzed by the enzyme phosphoglycerate
kinase:
1,3-bis-P-glycerate + ADP <---> 3-P-glycerate + ATP
is -18.8 kJ/mol at 37°C and pH 7.0. Knowing that in
yeast the ratio [ATP] / [ADP] = 10, calculate the
minimal ratio [3-P-glycerate] / [1,3-bis-P-glycerate] for
the reaction to work in the direction of 1,3 bis-P-
glycerate synthesis.
25. The oxidative decarboxylation of α-ketoglutarate
to succinate and CO2 through the Krebs cycle,
catalyzed by the α-ketoglutarate dehydrogenase
complex and the succinyl-CoA synthetase, with the
consequent transference of electrons to oxygen through
the mitochondrial respiratory chain, give rise to the
production of 3.5 ATPs. Calculate the efficiency of the
reaction under standard conditions. ∆G’o for the
synthesis of ATP = 30.5 kJ/mol.
ANSW ER KEY
1. a) ∆E'o = 0.15 V, b) NADH reduces acetaldehyde.
2. a) NADH reduces FMN; b) FMN / FMNH ; 2
c) NADH.
3. a) K’eq = 19.6; b) K’eq = 5.1x10-2; c) In the
direction of production of B+C
4. ∆G’o = -142.08 kJ/mol.
5. ∆G'o = -134.4 kJ/mol; Keq = 3.79 x 10 23.
6. a) Ethanol reduces dehydroascorbate;
b) ∆E’o = 0.26 V; ∆G’o = -49.92 kJ/mol.
7. a) Acetaldehyde reduces oxalacetate;
b) Acetaldehyde is oxidized and oxaloacetate is
reduced; c) ∆G’o = –82.56 kJ/mol;
d) K’eq = 3.05 x 10 .
14
8. a) Acetaldehyde reduces oxalacetate; b)
Acetaldehyde is oxidized; c) Oxalacetate is reduced;
d) ∆G’ = -79.10 kJ/mol.
9. a) NADH reduces pyruvate; ∆G' = -24.96 kJ/mol;
b) ~Equilibrium, ∆G’o = -0.38 kJ/mol; c) Lactate
reduces NAD , ∆G’o = -9.6 kJ/mol.
+
10. ∆G’o = -12.84 kJ/mol.
11. a) ∆G’ = -30.5 kJ/mol; b) ∆G’ = -36.2 kJ/mol;
c) ∆G’ = -41.9 kJ/mol; d) ∆G’ = -47.6 kJ/mol.
12. ∆G’ = -47.6 kJ/mol.
13. 0.00017 M.
14. a) ∆G’o = -14.63 kJ/mol; b) K’eq = 367.07.
15. a) ∆E’ = 0.25 V; b) ∆E’ = 0.625 V.
16. a) K’eq = 108.41; b) ∆Go’ = 18.9 kJ/mol.
17. ∆G’ = -49.31 kJ/mol.
18. a) K’eq = 199.8; b) K’eq = 5 x 10 .-3
19. a) ∆G’o = 2.09 kJ/mol; b) in the direction of
formation of glucose 6-P.
20. [glucose-1-P] = 4.6 x 10 M,
-3
[glucose-6-P] = 9.55 x 10 M -2
Classroom session III. Bioenergetics 3

21. E’o = -0.23 V.

22. a) ∆G’o = 4.46 kJ/mol; b) ∆G’ = -2.11 kJ/mol.
23. a) ∆G’ = -7.29 kJ/mol, in the direction of
formation of glucose-6-P; b) ∆G’ = 0 kJ/mol,
equilibrium; c) ∆G’ = 4.11 kJ/mol, in the direction
of formation of glucose-1-P.
24. [3-P glycerate] / [1,3 bis-P glycerate] > 198.72.
25. 37.32 %.

Degree in Veterinary Medicine. Biochemistry

Classroom session III. Bioenergetics 4

STANDARD REDOX POTENTIALS FOR SOME PAIRS OF INTEREST

(pH 7.0).
E’o (Vols)

1/2 O + 2 H + 2 --------------------------------------------->
2
+ e-
HO
2 +0.82
NO + 2e ----- ------------------------------------------------->
3
- -
NO 2
-
+0.42
Cit a (Fe ) + e ------------------------------------------------->
3+ -
Cit a (Fe ) 2+
+0.29
Cit c (Fe ) + e ------------------------------------------------->
3+ -
Cit c (Fe ) 2+
+0.22
Ubiquinone + 2H + 2e --------------------------------------> + -
Ubiquinone H 2 +0.10
Cit b (Fe ) + e ------------------------------------------------->
3+ -
Cit b (Fe ) 2+
+0.06
Dehydroascorbic acid + 2H + 2e ---------------------------> + -
Ascorbic acid +0.06
Fumarate + 2H + 2e ------------------------------------------>
+ -
Succinate +0.03
Standard 2H + 2e ---------------------------------------->
+ -
H2 0.00
Oxalacetate + 2H + 2e --------------------------------------> + -
Malate -0.17
FAD + 2H + 2e------------------------------------------------->
+
FADH 2 -0.18
Pyruvate + 2H + 2e ------------------------------------------>
+ -
Lactate -0.19
Acetaldehyde + 2H + 2e -----------------------------------> + -
Ethanol -0.20
Acetoacetate + 2H + 2e ------------------------------------> + -
β-Hidroxybutyrate -0.27
1,3 -BPG + 2H + 2e ---------------------------------------->
+ -
Glyceraldehyde-3-P + Pi -0.29
Lipoic acid + 2H + 2e --------------------------------------->
ox
+ -
Lipoic acid red -0.29
NAD + 2H + 2e ---------------------------------------------->
+ + -
NADH + H +
-0.32
NADP + 2H + 2e -------------------------------------------->
+ + -
NADPH + H +
-0.32
Acetyl~CoA + 2H + 2e -------------------------------------> + -
Acetaldehyde + CoA~SH -0.41
2H + 2e -------------------------------------------------------->
+ -
H2 -0.42
Acetate + 2H + 2e ----------------------------------------->
+ -
Acetaldehyde -0.60
Succinate + CO + 2H + 2e ----------------------------> 2
+ -
α-ketoglutarate + H O 2 -0.67

BASIC EQUATIONS IN BIOENERGETICS

ALL REACTIONS

ΔGo’ = -5.7 log K’eq

[Products]
ΔG’ = ΔGo’ + 5.7 log
[Substrates]

(ΔG’ = ΔG’o + 5.7 log q’ = -5.7 log (K’eq/q’)

REDOX REACTIONS

0, 06 [ox]
E = E 0ʹ + log
n [red]

ΔE’ = E’oxidant - E’reductor

ΔG’ = -96nΔE’
ΔG’o = -96nΔE’o

Degree in Veterinary Medicine. Biochemistry