Mulungushi University

School of Medicine and Health Sciences

MBChB program
Biochemistry MBI 261
2nd Year 1st semester TUTORIAL I
Prepared by Masenga SK., BSc., MSc., MSCI-CT
Total mark for this tutorial is 70
1. Consider the information given below for glycolysis reactions
no Reaction kJ/mol
1 Glucose + ATP → glucose-6-phosphate + ADP ΔGo = -16.7
2 Glucose-6-phosphate → Fructose-6-phosphate ΔGo = +1.7
3 Fructose- 6-phosphate + ATP → Fructose-1,6-diphosphate + ΔGo = -14.2
ADP
4 Fructose-1,6-diphosphate → glyceraldehyde-3-phosphate + ΔGo = +23.8
dihydroxyacetone phosphate
5 dihydroxyacetone phosphate → glyceraldehyde-3-phosphate ΔGo = + 7.5
6 glyceraldehyde-3-phosphate + phosphate + NAD+ →1,3- ΔGo = +6.3
diphosphoglycerate + NADH + H+
7 1,3-diphosphoglycerate + ADP → 3-phosphoglycerate + ATP ΔGo = -18.8
8 3-phosphoglycerate → 2-phosphoglycerate ΔGo = +4.6
9 2-phosphoglycerate → 2-phosphoenolpyruvate + H2O ΔGo = +1.7
10 2- phosphoenolpyruvate + ADP → pyruvate + ATP ΔGo = -31.4
11 pyruvate + NADH + H+ → lactate + NAD+ ΔGo = -25.1

Answer the following questions

a) Calculate the ΔGo of glycolysis (5 Marks)
Answer: -35.5kj/mol
b) State whether glycolysis is an exergonic or endergonic process and give a reason for your
answer (4 Marks)
Answer: Exergonic. The ΔGo is negative, the reaction proceeds spontaneously and
energy is released. Furthermore, there is a net ATP gain of 2
c) In class we learnt that almost all biochemical reactions including those of glycolysis are
coupled reactions (exergonic and endergonic). The Phosphorylation of glucose in reaction
(i) can be decomposed into two reactions.
Glucose + phosphate → Glucose-6-phosphate + H2O ΔGo = +14.3
ATP + H2O → ADP + phosphate . ΔGo = ????
What is the ΔG of the second reaction? State which one is endergonic and exergonic giving a
o

reason for each? (6 marks)

Answer:
ΔG0 = ∑ reaction 1 + reaction 2
• -16.7 = ΔGo of reaction 1 + ΔGo of reaction 2; -16.7 = +14.3 + ΔGo of reaction 2
ΔGo of reaction 2 = -16.7 – 14.3 = -31 kj/mol
• Glucose + phosphate → Glucose-6-phosphate + H2O ΔGo = +14.3 kj/mol ;
endergonic, ΔGo is positive
• ATP + H2O → ADP + phosphate . ΔGo = -31 kj/mol ; exergonic, ΔGo is negative

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 Mulungushi University
School of Medicine and Health Sciences
MBChB program
Biochemistry MBI 261
2nd Year 1st semester TUTORIAL I
Prepared by Masenga SK., BSc., MSc., MSCI-CT
d) The enzyme aldolase catalyzes reaction 4 (the conversion of fructose 1,6-diphosphate (FDP)
to dihydroxyactone phosphate (DHAP) and glyceraldehyde-3-phosphoate (GAP)). Under
physiological conditions the concentrations of these species in red blood cells (erythrocytes)
are [FDP] = 35 μM, [DHAP] = 130 μM and [GAP] = 15 μM at temperature 36.850C.
Will the conversion occur spontaneously under these conditions? (6 marks)
Solution: The standard free energy change for the reaction is
FDP → DHAP + GAP ΔGo = +23.8 kJ
So, since the reaction is not at equilibrium states, we will use the formulae; ΔG = ΔGo + RT
ln Q. we have to also change μM to M x 10-6

[DHAP][GAP]
Q = [FDP] = (130 x 10-6)(15 x 10-6)/ (35 x 10-6)
= 5.57 x 10-5

ΔG = ΔGo + RT ln Q
= 23800 J/mol + (8.31 J/mol-K)(310 K)ln(5.57 x 10-5)
= -1434 J/mol or -1.43 kJ/mol
The reaction will occur spontaneously under the conditions of the cell.

e) The coupled Reaction (i) can be divided into two half reactions as follows;
ATP + H2O <—> ADP + Pi ∆G0 = -31 kJoules/mol —> (1)
Pi + glucose <—> glucose-6-P+ H2O ∆G° = +14 kJoules/mol —> (2)
Which enzyme(s) catalyzes this reaction? Calculate the ∆G0 of the coupled reaction (4
marks)
Solution: Hexokinase or glucokinase (liver, pancreas)
Coupled reaction- (1) + (2)
ATP + glucose <—>ADP + glucose-6-P
∆G°’ = (-31 + 14) = -17 kJoules/mol

f)
i. What enzyme catalyzes reaction 3 and what two functions does ATP serve in this coupled
reaction? (6 marks)
Solution: The enzyme Phosphofructokinase 1 (PFK-1).
The hydrolysis of ATP provides the energy to drive this coupled reaction.
ATP also serves to donate a phosphate to fructose 1,6-bisphosphate from fructose 6-phosphate.

ii. Draw and label an energy diagram for this reaction. Include the relative energy levels of the
substrates and the products, the activation energy and the ΔG0 for the reaction. (6 marks)
Solution:

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 Mulungushi University
School of Medicine and Health Sciences
MBChB program
Biochemistry MBI 261
2nd Year 1st semester TUTORIAL I
Prepared by Masenga SK., BSc., MSc., MSCI-CT

iii. In erythrocytes, the following intracellular concentrations of metabolites are found:

Metabolite Concentration
Fructose 6-phosphate 0.014 mM
Fructose 1,6-bisphosphate 0.028 mM
AMP 1 mM
ADP 0.2 mM
ATP 2 mM
Pi 1 mM
What is the free energy change of the phosphofructokinase reaction under these cellular
conditions (37°C)? Show your work. Is the reaction spontaneous under these conditions? (6
marks)
Solution:
Change units for standard state free energy change to joules/mole; temperature into kelvins by
adding 273.15 to 370C and changing concentrations to moles by dividing by 1000.
ΔG = ΔGo + RT ln Q
[𝐹𝑟𝑢𝑐𝑡𝑜𝑠𝑒 1,6−𝑏𝑖𝑠𝑝ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒][𝐴𝐷𝑃]
ΔG = ΔGo + RT ln [𝐹𝑟𝑢𝑐𝑡𝑜𝑠𝑒 6−𝑝ℎ𝑜𝑠𝑝ℎ𝑎𝑡𝑒][𝐴𝑇𝑃]
[0.000028][0.0002]
ΔG = - 14200 j/mol + 8.314 X 310.15 In [0.000014][0.002]
ΔG = - 14200 j/mol - 4148.9
ΔG = -18348.9 j/mol OR -18.3489 KJ/mol.
The reaction will proceed forward; spontaneous

g)
i. Which enzymes regulate glycolysis? (3 marks)

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 Mulungushi University
School of Medicine and Health Sciences
MBChB program
Biochemistry MBI 261
2nd Year 1st semester TUTORIAL I
Prepared by Masenga SK., BSc., MSc., MSCI-CT
Hexokinase (and glucokinase), phosphofructokinase-1, pyruvate kinase

ii. What specific signals or molecule(s) do these enzymes respond to? (6 marks)
enzyme activator inhibitor
Hexokinase Glucose Glucose 6 phosphate inhibitor
(but not glucokinase)
Glucokinase ↑carb feeding induced by ↓fasting repressed by glucagon
insulin
Phosphofructokinase-1 5’AMP, fructose-6- citrate, ATP, glucagon, Low PH
phosphate, fructose 2,6-
bisphosphate, Pi, insulin;
Pyruvate kinase fructose 1,6 bisphosphate, ATP, alanine, glucagon,
insulin; norepinephrine

iii. Under what regulation mechanisms are they? (2 marks)

Allosteric and feedback control
iv. Why is glycolysis regulation important? (2 marks)
Solution: To help provide energy as and when needed by cell. Normally, a low ATP to ADP
ratio indicates that cells need more energy currency available ready to use. By speeding up
glycolysis, more ATP can be formed from ADP to raise the ATP to ADP ratio.

h) Under what oxygen conditions do these reactions (i to ii) proceed to and if the condition
was the opposite to the one suggested in the final reaction fate of glycolysis product, what
would be the product and enzyme catalyzing the very next reaction under normal
conditions ? (3 mark)
Solution: anerobic condition; the opposite is aerobic condition to which the reaction fate of the
product of glycolysis (pyruvate) would be acetyl-CoA catalized by Pyruvate dehydrogenase
complex
i) In certain tumor cells, an enzyme called ATPase becomes abnormally active, resulting in
increased hydrolysis of ATP to ADP. What would be the effect on the overall rate of
glycolysis in these cells and explain the biochemical mechanism behind cachexia (wasting &
weakness) in chronic cancer patients? (5 marks)
Solution: The activity of the glycolytic enzymes would increase, increasing the overall rate of
glycolysis.
High rate glycolysis (more than oxygen supply) → lactate → gluconeogenesis → great energy
expense (loss energy) → cachexia
j) The harvested energy in glycolysis is stored in what molecule(s)? and what is/are these
molecule(s) used for? (6 marks)
Solution: NADH and ATP. ATP is used to drive reactions in the cell, and NADH is used to
regenerate NAD+ for the next rounds of glycolysis.

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