Joice Anne U.

Tolosa

1. Bacteria, like all living organisms, needs energy to survive. All living organisms need
ways to get nutrients and energy, including bacteria. Cells are able to acquire energy in
a wide variety of ways, including cellular respiration. Cellular respiration is a process that
uses oxygen, nitrate, or sulfate to break down nutrients to generate a cell's energy. If
oxygen is used, it is called aerobic cellular respiration. If oxygen is not used, it is
called anaerobic cellular respiration. Cells, including bacteria, can be thought of as
energy producing factories that take nutrients and convert them into energy called
adenosine triphosphate (ATP). Some cells may be better than others at producing ATP
because they use more efficient methods. The benefit of using cellular respiration for
bacteria is the amount of energy or ATP generated. The reason cellular respiration
generates so much ATP is because it maximizes the use of glucose by using by-
products generated in other energy producing pathways.
Cells couple the exergonic reaction of ATP hydrolysis with endergonic reactions to
harness the energy within the bonds of ATP.

ATP is hydrolyzed into ADP in the following reaction:

ATP+H2O→ADP+Pi+free energy

Like most chemical reactions, the hydrolysis of ATP to ADP is reversible. The reverse reaction
combines ADP + Pi to regenerate ATP from ADP. Since ATP hydrolysis releases energy, ATP
synthesis must require an input of free energy.

ADP is combined with a phosphate to form ATP in the following reaction:

ADP+Pi+free energy→ATP+H2O

ATP and Energy Coupling

Exactly how much free energy (∆G) is released with the hydrolysis of ATP, and how is that free
energy used to do cellular work? The calculated ∆G for the hydrolysis of one mole of ATP into
ADP and Pi is −7.3 kcal/mole (−30.5 kJ/mol). However, this is only true under standard
conditions, and the ∆G for the hydrolysis of one mole of ATP in a living cell is almost double the
value at standard conditions: 14 kcal/mol (−57 kJ/mol).

ATP is a highly unstable molecule. Unless quickly used to perform work, ATP spontaneously
dissociates into ADP + Pi, and the free energy released during this process is lost as heat. To
harness the energy within the bonds of ATP, cells use a strategy called energy coupling.

Energy Coupling in Sodium-Potassium Pumps

 Energy Coupling: Sodium-potassium pumps use the energy derived from exergonic ATP
hydrolysis to pump sodium and potassium ions across the cell membrane.

Cells couple the exergonic reaction of ATP hydrolysis with the endergonic reactions of cellular
processes. For example, transmembrane ion pumps in nerve cells use the energy from ATP to
pump ions across the cell membrane and generate an action potential. The sodium-potassium
pump (Na+/K+ pump) drives sodium out of the cell and potassium into the cell. When ATP is
hydrolyzed, it transfers its gamma phosphate to the pump protein in a process called
phosphorylation. The Na+/K+ pump gains the free energy and undergoes a conformational
change, allowing it to release three Na+ to the outside of the cell. Two extracellular K+ ions bind
to the protein, causing the protein to change shape again and discharge the phosphate. By
donating free energy to the Na+/K+ pump, phosphorylation drives the endergonic reaction.

Energy Coupling in Metabolism

During cellular metabolic reactions, or the synthesis and breakdown of nutrients, certain
molecules must be altered slightly in their conformation to become substrates for the next step
in the reaction series. In the very first steps of cellular respiration, glucose is broken down
through the process of glycolysis. ATP is required for the phosphorylation of glucose, creating a
high-energy but unstable intermediate. This phosphorylation reaction causes a conformational
change that allows enzymes to convert the phosphorylated glucose molecule to the
phosphorylated sugar fructose. Fructose is a necessary intermediate for glycolysis to move
forward. In this example, the exergonic reaction of ATP hydrolysis is coupled with the
endergonic reaction of converting glucose for use in the metabolic pathway.

2 flasks of e.coli are grown in batch culture in the samemedium(2%glucose and amino acids; no
nitrate) and the sametemperature(37c). Culture 1 is well aerated. Culture 2 is anoxic.After 16
hours the following observations are made:

 culture #1 has a high cell density; the cells appear to be in astationary phase and the
glocose level in the medium is reduced to1.2%
 culture #2 has a low cell density; the cells appear to be inthe logarithmic phase, although
their doubling time isprolonged(over one hour). The glucose level is reduced to0.2%.

what type of glucose catabolism was used in each culture? whydoes culture 2 have so little
glucose remaining relative to culture#1, even though culture #2 displayed slower growth and
has lessbiomass?

Answered
 Joice Anne U. Tolosa Cell Biology

MAEd Biology 1

1. From an evolutionary perspective, discuss why most microorganisms use aerobic

respiration to generate ATP. Expalin how ATP performs cellular work.
Answer:
Bacteria, like all living organisms, needs energy to survive. All living organisms need
ways to get nutrients and energy, including bacteria. Cells are able to acquire energy in
a wide variety of ways, including cellular respiration. Cellular respiration is a process that
uses oxygen, nitrate, or sulfate to break down nutrients to generate a cell's energy. If
oxygen is used, it is called aerobic cellular respiration. If oxygen is not used, it is
called anaerobic cellular respiration. Cells, including bacteria, can be thought of as
energy producing factories that take nutrients and convert them into energy called
adenosine triphosphate (ATP). Some cells may be better than others at producing ATP
because they use more efficient methods. The benefit of using cellular respiration for
bacteria is the amount of energy or ATP generated. The reason cellular respiration
generates so much ATP is because it maximizes the use of glucose by using by-
products generated in other energy producing pathways.
Cells couple the exergonic reaction of ATP hydrolysis with endergonic reactions to
harness the energy within the bonds of ATP.

2. Two flasks of e.coli are grown in batch culture in the samemedium(2%glucose and
amino acids; no nitrate) and the sametemperature(37c). Culture 1 is well aerated.
Culture 2 is anoxic.After 16 hours the following observations are made:

 culture #1 has a high cell density; the cells appear to be in astationary phase and the
glocose level in the medium is reduced to1.2%
 culture #2 has a low cell density; the cells appear to be inthe logarithmic phase, although
their doubling time isprolonged(over one hour). The glucose level is reduced to0.2%.

what type of glucose catabolism was used in each culture? whydoes culture 2 have so little
glucose remaining relative to culture#1, even though culture #2 displayed slower growth and
has lessbiomass?

Culture 1.

 It is well aerated.
 Develops high cell density.
 Glucose level reduced to 1.2%
 Type of glucose catabolism used is aerobic.

Aerobic catabolism supplies the cell the 12precursor molecules, reducing power (NADH &
NADPH), & ATP.Of these, 6 are produced by Glycolysis, tricarboxylic acid cycle(TCA) produces
four, and pentose phosphate pathway producestwo.

This catabolic process involvesoxidation/reduction processes involving electron transfer. In

thistype of catabolic process (aerobic), ATP is produced bychemiosmosis, and creates 12
precursor molecules. Reducing power isstored in NADH and NADPH.

First the monosaccaride Glucose is split, andconverted into two pyruvates. ATP is initially
required in highquantitiy, but later only in small quantity. A small amount of NADHis also
produced. Subsequently in the Kreb's cycle, some pyruvateis used in biosynthesis, and some is
oxidised to end up ascitrate.In a series of six reactions, CO2 is released andoxalocitrate is
regenerated. This is the catabolic part in aerobicmetabolism.

In high growth rate cultures as this one, ATPusage is reduces as ATP is a fragile molecule and
has to be madeagain and again. So, in culture 1, 1.2% is left.

Culture 2.

 It is Anoxic.
 Develops low cell density.
 Glucose level is only 0.2%
 Type of glucose catabolism used is anaerobic.

Anaerobic catabolism also involves an electrontransfer chain, but without oxygen. However if
the process involves fermentation, no electron transfer chain is involved. Also, the ATP released
per glucose molecule is three in anerobic catabolism to 38 in aerobic.

In this culture, growth rate is slower. So, theATP burden is increased as cell maintenance load
increases due tolonger time to replicate. This leads to increased and inefficientGlucose usage
(only three molecules of atp/glucose molecule). Thisleaves only 0.2% glucose in the end.

Aerobic catabolism makes efficient usage ofglucose, and more cells are built in a short time.
leaving greaterquantity of glucose unused.

Anaerobic catabolism is much more inefficient inusing glucose and also use more glucose as
the unstable ATPmolecule has to be rebuilt again and again over the longer time thecell takes to
replicate, to maintain it. This utilises more of theglucose, has a thinner cell density, and longer
replication time,and leaves back less glucose.