1.

We learned that the enzyme RuBisCo performs carbon fixation by using CO2 as a
substrate. However, it can also use O2 as a substrate. If oxygen is used instead of
carbon dioxide, what do you expect the reaction product(s) to be?
Draw the reactants and product(s) for both scenarios (use of CO2 and use of O2)

The reaction with carbon dioxide cleaves the bond between the 2nd and 3rd carbons of
ribulose-1,5-bisphosphate and incorporates the additional carbon from the CO2 to
generate 2 molecules of 3-phosphoglycerate (3PG).
We can expect that oxygen will interact at the same location, but will not be used to
incorporate an additional carbon, so we end up with 1 molecule of 3PG and 1 molecule
of 2-phosphoglycerate (2PG).

2. Even though the Calvin cycle is considered a light-independent pathway, it is still

regulated by light. Explain the ways in which this occurs.

There are two regulation mechanisms that occur due to the presence of light.

RuBisCO has an optimal function at a pH of 8 in the stroma. In chloroplasts, H+ is either

produced by the oxidation of water in PSII or pumped into the thylakoid lumen by Q and
Cytochrome b6. To balance the electrochemical shift, Mg2+ travels through a channel
into the stroma during the light-dependent reactions of photosynthesis, which allows it to
stimulate RuBisCO. Thus, the presence of Mg2+ and pH of the stroma that RuBisCO
relies on is dependent on light-determined activation.

Bisphosphatases are also regulated by light, as light allows PSI to reduce Fd, which
initiates a series of reduction reactions that eventually reduces and activates
Bisphosphatases.

3. Determine whether the statements below are True or False.

a) The enzyme RuBisCo fixes carbon dioxide in __False__

a reaction with the ketose ribulose-5-phosphate.
Rubisco catalyzes this reaction with ribulose-1,5-bisphosphate.
Ribulose-5-phosphate must first be phosphorylated by
Phosphoribulokinase to generate ribulose-1,5-bisphosphate.
 b) Each full turn of the Calvin cycle generates 6 molecules of __False__
glyceraldehyde-3-phosphate (GAP) which are used to
synthesize 2 molecules of carbohydrates in the form of glucose.
Only one GAP is siphoned off for use in carbohydrate synthesis.
The other 5 molecules of GAP are used to regenerate 3 molecules
of ribulose-1,5-bisphosphate (RuBP).
c) Transketolase reactions transfer 2 carbon units at a time and __True__
use the cofactor TPP.

d) In the Calvin cycle, only the phosphoribulokinase and __True__

bisphosphatase reactions are irreversible.

e) Bisphosphatase is activated by the oxidation of reduced __True__

thioredoxin, which is regenerated by the oxidation of
ferredoxin-thioredoxin reductase.
f) The net reaction of the pentose phosphate pathway (PPP) is __False__
3 Glucose-6-phosphate + 6 NADPH NADP+ + 3 H2O →
→ 3 CO2 + 6 NADP+ NADPH + 6 H+ + 2 Fructose-6-phosphate + GAP
Although 6 protons are generated in the net reaction, NADP+
is consumed and NADPH is generated, so these are flipped.

4. Consider two different populations of cells

a) Based on the changes in substrate concentration over time, what does the flux through
the pentose phosphate pathway look like for population A?
Draw the pathway and include the names of the major substrates involved. Explain your
answer.

Because there is an increase in both

ribose-5-phosphate and NADPH, that must mean
that glucose-6-phosphate is being diverted from
glycolysis to travel through the oxidative pathway to create NADPH and ribulose-5-phosphate,
which is then converted to ribose-5-phosphate, to be used for alternative purposes like
nucleotide synthesis, rather than continuing through the non-oxidative pathway. This is
confirmed by the decrease in glucose-6-phosphate that would be expected of the pathway that
requires glucose-6-phosphate as the first reactant.

b) Based on the changes in substrate concentration over time, what does the flux through
the PPP largely look like for population B?
Draw the pathway and include the names of the major substrates involved. Explain your
answer.

Only NADPH concentrations significantly increase

over the time period given. This means that the
oxidative phase is upregulated to create NADPH.
However, glucose-6-phosphate is not decreasing,
meaning that there must be some way the cycle is
replenishing concentrations. One way to accomplish
this is by using the product of the oxidative pathway,
ribose-5-phosphate, to create non-oxidative pathway
products fructose-6-phosphate and
glyceraldehyde-3-phosphate. These intermediates
can be diverted back into glycolysis to then replenish
glucose-6-phosphate through gluconeogenesis.

Note: Don’t study gluconeogenesis for Exam 3. Just

understand that the non-oxidative phase can be
diverted back to glycolysis to replenish reactants.

c) Using the substrate concentrations of populations A and B, determine which population

would be best able to respond to a rapid increase in oxidative stress. Explain why.

Population B would be best equipped to handle oxidative stress. Population B has the highest
concentration of NADPH abundance compared to other reactants. NADPH can be used to
regenerate glutathione in the reaction catalyzed by glutathione reductase. Glutathione is a
cellular weapon against damaging reactive oxygen species (ROS) that are created from high
levels of oxidative stress. Thus, population B could reduce the most glutathione to aid in
combating the ROS.
 d) Based on the difference in substrate concentrations, which population’s growth rate is
the highest? Explain your answer.

Ribose-5-phosphate can be diverted from the pentose phosphate pathway to synthesize

nucleotides. Nucleotides are needed in higher concentrations for rapidly dividing cells that must
constantly replicate DNA and generate two cell bodies from one. Population A showed the
greatest increase in ribose-5-phosphate, indicating that nucleotide synthesis in that population is
likely higher than in population B. Using this information, we can then conclude that population
A divides more rapidly than population B.