Mulungushi University School of Medicine and Health Sciences

Biochemistry... 2022/2023
Carbohydrate Metabolism Recap 2 Solutions

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 + ADP ΔGo = -14.2
4 Fructose-1,6-diphosphate → glyceraldehyde-3-phosphate + dihydroxyacetone phosphate ΔGo = +23.8
5 dihydroxyacetone phosphate → glyceraldehyde-3-phosphate ΔGo = + 7.5
6 glyceraldehyde-3-phosphate + phosphate + NAD+ →1,3-diphosphoglycerate + NADH + H+ ΔGo = +6.3
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 (2 Marks)
Ans: Answer: -35.5kj/mol (Only sum up the first 10 reactions)

b) State whether glycolysis is an exergonic or endergonic process and give a reason for your
answer (2 Marks)
Ans: 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 (1)
can be decomposed into two reactions.
1. Glucose + phosphate → Glucose-6-phosphate + H2O ΔGo =+14.3
2. ATP + H2O → ADP + phosphate. ΔGo =????

What is the ΔGo of the second reaction? State which one is endergonic and exergonic giving
a reason for each? (2 marks)

Ans:
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

d) What enzyme catalyses reaction 3 and what two functions does ATP serve in this coupled
reaction? (3 marks)
Ans: Phosphofructokinase-1
Ans: ATP is used for:
• ATP also serves to donate a phosphate to fructose 1,6-bisphosphate from
fructose 6-phosphate.
• The hydrolysis of ATP provides the energy to drive this coupled reaction.

e) Which enzymes regulate glycolysis? (3 marks)

Ans: Hexokinase/Glucokinase, Phosphofructokinase and Pyruvate kinase.

f) What enzyme catalyzes reaction 11 and under what conditions? (2 marks)

Ans: Lactate Dehydrogenase.
Ans: Conditions;
• Anaerobic conditions

g) In certain tumour 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? (3 marks)
Ans: Accumulation of ADP increases the rate of glycolysis as ADP stimulates glycolytic
regulatory enzymes.
Ans: High-rate glycolysis (more than oxygen supply) → lactate → gluconeogenesis →
great energy expense (loss energy) → cachexia

2. Show and name the three bonds in ATP between the phosphates and the sugar? (2 Marks)
Also show α, ß, γ phosphates and state which one undergoes hydrolysis to yield energy (4
marks)

Ans: Gamma phosphate group undergoes hydrolysis.

a) List the two main sites of glycogenesis and the enzyme (s) that regulate this process?
(3 marks):
b) Ans: Sites are; Liver and Muscle.
Ans: Regulated by; Glycogen synthase

3. Under what conditions is Glycogenolysis activated and for what purpose? 6 marks
Ans:
Starvation
Fasting
Low carbohydrate diet
Intense exercise
 Glucagon Presence
In between meals

4.
a) What are the phases and key products of the hexose monophosphate shunt and what are
these products used for? (6 marks)
Ans: Oxidative Phase (Irreversible) and No-Oxidative phase (Reversible)
Ans: Ribose-5-Phosphate = Used to make DNA and RNA
= Used to make NADPH molecules which help in building
other molecules.
Ans: Xylulose-5-Phosphate = Has a role in gene expression
= Is involved in glycolysis as it is converted into Fructose-6-
Phosphate.

b) List at least 4 key differences between Glycolysis and the pentose phosphate pathway 4
Marks

5.
a) What are the key Differences between Fructolysis and glycolysis 5 marks
Fructose:
•Insulin dependence: No
•Transporters: GLUT 5
•Rate of metabolism: Rapid, bypass rate limiting step PFK-1
•Sites: Liver, kidney, intestines, brain
•Purpose: Replenish Liver glycogen & TAG synthesis

b) Explain the mechanism of the metabolic consequences of large fructose quantity Ingestion
responsible for atherosclerosis and the weak feeling that accompanies sucrose/HFCS
ingestion in some people? 5 marks
Ans: Fructolysis leads to the production of triglycerides which are lipids. Over
production of lipids may lead to them being deposited in the lumen of blood vessels
which can blood the blood vessels, hence, causing atherosclerosis.

6. A patient suspected of T2DM came to the hospital. Urine was collected and a blood
sample. What was the urine for? What tests can you conduct in a blood specimen to diagnose
prediabetes and T2DM? what are the reference ranges 11 marks

7. Dinitrophenol (DNP) is an uncoupler, or has the ability to separate the flow of electrons
and the pumping of H+ ions for ATP synthesis. This means that the energy from electron
transfer cannot be used for ATP synthesis. Fifty years ago, DNP was given as a drug to help
patients lose weight. Why does this work? Why would this be dangerous? 5 marks
Ans: DNP causes weight loss by making cellular respiration less efficient. The majority
of ATP (molecules used for energy) is produced in the mitochondria. ... Your body then
needs to continue to burn more carbs, fat, and protein at a higher rate to make up for
the shortage of ATP. If electron transport doesn't produce ATP, then much more sugar
must be metabolized for energy needs. Very low production of ATP would be lethal.
Energy usually stored as ATP molecules becomes dissipated as heat, potentially leading
to clinically significant or even lethal hyperthermia.

In oxidative phosphorylation, the flow of electrons from NADH and FADH2 to oxygen
results in the pumping of H+ from the matrix to the inner membrane space. This
gradient of H+ can produce ATP by flowing through ATP synthetase in the
mitochondrial inner membrane. Dinitrophenol disrupts the H+ gradient reducing ATP
synthesis. Under these conditions, much of our food that we eat could not be used for
ATP synthesis and we lose weight. However, too much inhibitor and we could make too
little ATP for life. The difference between weight loss and death is only a small
concentration change in dinitrophenol, making the drug dangerous.

8. If you isolate mitochondria and place them in buffer with a low pH they begin to
manufacture ATP. Why? 3 marks

Ans: The high external acid concentration causes an increase in H+ in the inter
membrane space leading to increased ATP production by ATP synthetase.
Mitochondrial production of ATP requires a concentration gradient of H+, with a high
concentration at the inter membrane space and a low concentration in the matrix. The
inner membrane is impermeable to H+, but the outer membrane of the mitochondria
will allow H+ to pass through. Thus, placing mitochondria in a low pH buffer produces
a H+ gradient that can generate ATP through ATP synthetase.

9. List the four Stages of respiration (per mol of glucose under aerobic conditions to yield
energy) and the number of ATPs and electron carriers (H+) generated per stage. Then finally
show the total ATP formation (10 marks)

Ans: 4 stages of energy formation (cellular respiration)

1. Gycolysis (Glucose → 2 Pyruvate) yields 2 net ATPs
2. Preparatory stage: Pyruvate dehydrogenase complex (2 Pyruvate → Acetyl CoA)
yields 2 NADH
3. Krebs cycle x 2 → 2 ATP
4. Electron transport Chain:
2 NADH from glycolysis (malate shuttle) yields 5ATP
2 NADH from preparatory stage yields 5ATP
Krebs - 6 NADH (6 x 2.5 ATPs=15 ATP) ,2 FADH2 (2 x 1.5 ATP=3 ATP)
Total 18 ATPs
Total 28
Total for all stages 32.

Note: Most Biology text books like to round off the ATPs generated from NADH
(2.5~3ATPs) and FADH2 (1.5~2ATPs) in the ETC for making calculations appear much
simpler. This means 6 ATPs (from stage 1); 6 ATPs (stage 2) ;22 ATPs (Krebs) - 6
NADH (6 x 3 ATPs=18 ATP), 2 FADH2 (2 x 2 ATP=4 ATP) Total 34 ATPs from ETC,
hence:
TPs from ETC + 2ATPs (glycolysis) + 2ATPs Krebs = 38 ATPs. This assumes
malate/aspartate shuttle. Using the glycerol phosphate shuttle, we yield 36 ATPs

10. Calculate the metabolic efficiency of Glycolysis and of the complete metabolism of 1
mole of glucose in the ETC? also state the purpose of the deficit (100% ─ efficiency). 4
marks
Note: One mole of Glucose = 2805 kJ/mole and 1 ATP is equivalent to 30.5 kJ/mole

Ans:
32 ATP are produced per glucose (glycolysis → ETC) = 32 ATP x 30.5 kJ/mole = 976
kJ/mole
976 divided by 2805 X 100%= 35%
Efficiency of total glucose metabolism = 35%
65 percent is lost as heat. In humans, this heat assists in maintaining body temperature.

Glycolysis efficiency
Glucose = 2805 kJ/mole
Glycolysis = 2 ATP x 30.5 kJ/mole = 71 kJ/mole
71 kJ/mole divided by 2805 = 2.5% efficiency

11. You set up an experiment under aerobic condition and test how much ATP is made in
cells in the presence of an inhibitor. You find that from 3 moles of glucose, only 51 moles of
ATP are generated.
a. How many mole moles of ATP would you expect to generate in the absence of the
inhibitor? Show your
work 5 marks
b. Name the enzyme in the TCA cycle that was affected by adding the inhibitor? Show your
work 10 marks
Glycolysis
• 2 ATPs X 3 moles glucose = 6 ATPs
• 2 NADH= 5ATP X 3 moles of glucose = 15 ATPs
Total 21 ATPs at this stage

2 Pyruvate→ acetyl CoA (preparatory):

• 2 NADH= 5ATP X 3 moles of glucose = 15 ATPs
Total 36 ATPs at this stage
TCA cycle
• Reaction by isocitrate dehydrogenase 2 NADH= 5ATP X 3 moles of
glucose = 15 ATPs
Total 51 ATPs at this stage
Since we have the 51 ATPs that were generated, this means the next enzyme in the TCA
cycle generating a reducing equivalent is the enzyme that is inhibited. This is α-
ketoglutarate .
11 B Hint: (dehydrogenase complex)

“The power to perform miracles lies up within us and up above us.”

-Silz