Analyzing the Effect of Salt Concentration Levels on Height of Oxygen

Bubbles Produced from Catalase in Yeast

By: Meera Kumar

AP Biology - Period 4
Mr. David Robertson

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1) INTRODUCTION:
From pH to the presence of different modulators, all enzymes have different optimal conditions where
they can function best. In this experiment, we took a closer look at the enzyme catalase and how different salt
concentrations affect its ability to perform. Catalase is an enzyme that’s purpose is to break hydrogen peroxide
up into water and oxygen. The way we tested this was by activating yeast, an organism in which this enzyme is
present, in solutions of different salt concentrations of 0%(control group), 0.5%, 1%, 2%, and 3%, our
independent variables. We then dropped a small amount of the yeast solution into hydrogen peroxide and
measured the functionality of the catalase. To do this we measured the height of the oxygen bubbles in
millimeters, as oxygen is one of the products of this reaction; this served as the dependent variable for our
experiment. Some of the controls we had included the amount of hydrogen peroxide used, the amount of yeast
solution used, and the environment in which the experiment was taking place. From what we know about
catalase, it could be expected to see an increase in oxygen production at first, and then after that a very dramatic
decrease. This is because catalase is usually found in environments with some sort of salt concentration, so at
first we would be approaching its optimal condition; however, after it was in an environment with too much
salt, the enzyme would start to lose its ability and would eventually denature. I predicted that this peak would
happen somewhere between 0.5% and 1.0% because that is around the salt concentration range that many
catalase-containing cells have.
2) DATA TABLES:

Data Table #1: Oxygen Produced from Yeast Activated in Various Salt Concentrations
Salt Concentration Height of Bubbles Height of Bubbles Reaction
Level after 0 sec(mm) after 10 sec(mm) Rate(mm/sec)

0.0% NaCl 0 4 0.4

0.5% NaCl 0 3 0.3

1.0% NaCl 0 23 2.3

2.0% NaCl 0 6 0.6

3.0% NaCl 0 5 0.5

Data Table #2: Class Average for Rates of Reaction

0.0% 0.5% 1.0% 2.0% 3.0%
NaCl(mm/sec) NaCl(mm/sec) NaCl(mm/sec) NaCl(mm/sec) NaCl(mm/sec)

0.43 0.54 0.37 0.5 0.34

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3) GRAPHS:

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4) ANALYSIS:
From this data, we are able to see that the catalase from the reaction produced the most oxygen bubbles
around 0.5% salt concentration, and after reaching its optimum, the values started to decrease. This trend
confirms the initial hypothesis that after the reaction reaches its peak in terms of the oxygen produced it will
decrease; however, the decrease after the graph peaked was much less drastic than I initially thought it may
have been. For example, when looking at the class average data, the average rate of change from 0% to 0.5% is
increasing by 0.22 mm/sec, while from 0.5% to 3%, the average rate of change is decreasing by only a mere
0.08 mm/sec. Also, in the class average data, we see another small peak at 2% salt concentration, which may be
due to an error, as we had to discard two measurements that were outliers in that particular sample. In the
individual data, that outlier is very visible at the 1% salt concentration; from 0.5% to 1% we witnessed an
increase of around 666.67% in terms of the rate of change, and from 1% to 2%, there was a decrease by about
73.91%. This could be due to a number of potential factors, the most plausible being that a clump of yeast was
also sucked into the pipette before the solution was dropped into the hydrogen peroxide. One way that this
could possibly be avoided in the future is by straining out the solution so no clumps are able to get through.
Nevertheless, even with this possible error, the same trend persisted: after the highest amount of oxygen bubbles
are produced, less and less start to be produced afterwards. From this trend, we are able to draw the conclusion
that catalase is most effective at around 0.5% to 1% salt concentration. This also can be connected back to the
salt concentration in human cells, which sits at around 0.7-0.9% salt concentration. Since our cells also use
catalase, it would make sense that their optimal condition in terms of salt concentration is similar to that of the
one they have to function in. Furthermore, when you look at the experimental groups in comparison to the
control of 0% salt concentration, it is visible that almost all of the values of O2 bubbles produced are still higher,
even if just by a little bit. This means that the catalase was able to function better in an environment with a salt
concentration much higher than its ideal rather than an environment with no salt at all, leading to another
question: at what percentage of salt concentration does no salt become more effective? and when does catalase
denature? Overall, even though there were some deviations from the overall trends, after having completed this
experiment, the importance of enzymes functioning in their environments close to their optimums is very clear.