Sanya Daryani 10B Enzyme Activity Lab April 26, 2019

Name: Sanya Daryani

Subject: Biology
Submission Date: April 26, 2019
Investigation Title: Investigating a factor affecting enzyme activity - Criterion C
SOI: Each component in a system perform its specific function at the right time and place for the
system as a whole to be successful

Research Question:

How does Increasing the Temperature (10°C, 20°C, 30°C, 40°, 50C° ±0.5°C) from which the

Enzyme catalase from mushrooms is exposed to affect the rate of decomposition (mL/min) of hydrogen

peroxide measured by the time taken to form 10 mL (±0.1 mL) of foam?

Hypothesis:

If the temperature (10°C, 20°C, 30°C, 40°C, 50°C, ±0.5°C)

of the catalase in the mushrooms which is submerged into hydrogen

peroxide (H2O2) and detergent increase, then the H2O2 will

decompose at a faster rate. When a high temperature is presented,

the heat causes molecules to vibrate and have more space to move

freely amongst themselves. Furthermore, as temperature rises, more

kinetic energy will be present within reacting molecules. Also,

Figure 1 - optimum temp. graph
because of the high amount of energy that is produced, the “Enzymes.” Chemistry for Biologists: Enzymes,
Nov. 2004, www.rsc.org/Education/
molecules reach its activation energy which then creates a collision
Teachers/Resources/cfb/
between the catalase and hydrogen peroxide. This results in a fast

rate of reaction as well as the enzyme activity. When the temperature of the catalase is lowered, the rate of

decomposition for the H2O2 to react with catalase takes longer, leading to the 10 mL of foam to produce at a

slow pace. However, enzymes have an optimal temperature and this is the temperature where they best

facilitate reactions. The optimum temperature usually ranges between 35°C and 40°C, giving an average of

37°C. Due to optimum temperature, once an enzyme has reached its peak, which is 37°C, they will denature

and will cause the rate of decomposition to become slower. This is because high temperatures cause

intramolecular and intermolecular bonds to break, leading to the enzyme gaining more energy. In figure 1,

the expected graph illustrates that as the temperature increases, the enzyme activity increases too until it

reaches 37°C, and slowly starts to break down since higher levels of temperature causes rapid rates of

!1
Sanya Daryani 10B Enzyme Activity Lab April 26, 2019

denaturation. Hence, figure 1 displays an exponential curve, gradually increasing, gets to its highest point

(optimum temperature) and then decreases at a very steep rate.

Raw Data:

Figure 2 - Raw data for the rate of decomposition by measuring the time taken to produce 10 mL of foam

The Effect of Increasing the Temperature of Enzyme Catalase from Mushrooms on the rate of
Decomposition of the Substrate H2O2
IV
The Temperature of DV
catalase enzyme from The Rate of Decomposition o H2O2 measured by the time taken to produce
mushroom (°C) 10 mL of foam
Uncertainty: ±0.5°C Uncertainty: ±0.01 min

Trial 1 Trial 2 Trial 3

10 8:21 21:08 5:32

20 8:25 14:10 15:08

30 12:45 9:06 13:51

40 17:51 9:31 11:01

50 15:28 10:08 14:47

!2
Sanya Daryani 10B Enzyme Activity Lab April 26, 2019

Processed Data Table:

Figure 3 - Data for the effect of increasing temperature of catalase enzymes on the rate of decomposition of

the substrate H2O2

The Effect of Increasing the Temperature of Catalase Enzyme from Mushrooms on the rate of
Decomposition of the Substrate H2O2

DV
IV The Rate of Decomposition o H2O2 measured by the time taken to
The Temperature of catalase produce 10 mL of foam
enzyme from mushroom (°C) Uncertainty: ±0.01 min
Uncertainty: ±0.5°C
Time Rate Average Rate Standard
(1/time) Deviation
Trial 1 8.35 0.12

10 Trial 2 21.35 0.05 0.12 0.07

Trial 3 5.53 0.18

Trial 1 8.42 0.12

20 Trial 2 13.41 0.07 0.09 0.03

Trial 3 15.13 0.07

Trial 1 12.75 0.08

30 Trial 2 9.10 0.11 0.09 0.02

Trial 3 13.85 0.07

Trial 1 17.85 0.06

40 Trial 2 9.52 0.11 0.09 0.03

Trial 3 11.02 0.09
Trial 1 15.47 0.06

50 Trial 2 10.13 0.10 0.08 0.02

Trial 3 14.78 0.07

Sample Calculations:
Finding the conversion of minutes to seconds to minutes for 10°C Trial 1
8:21 mins (out of 60 seconds)
8 x 60 = 480 sec
480 + 21 = 501 sec
501 / 60 = 8.35 min (hundreths place)

!3
Sanya Daryani 10B Enzyme Activity Lab April 26, 2019

Finding the rate of reaction for 10°C Trial 1

1/501 = 0.12 mL/min

Finding the average rate for 20°C all trials

(0.12 + 0.07 + 0.07) / 3 = 0.09 mL/min

Finding the standard deviation for all trials

Qualitative Observations:

This image shows three trials of the 20°C increment being executed. In

each of the graduated cylinders, white foam is forming at slow paces.

The hydrogen peroxide and detergent were clear and colourless, while

the mushrooms were dark brown and murky. Because of the visibility

of the solution and uneven rising of the foam, the observations made

could have affected the results due to the possible errors that could

occur throughout the experiment. The observations could have

contributed to inconsistencies shown on the raw data table (figure 2) and eventually led to the lack of

reliability of data shown on the graph (figure 4). Throughout the lab report, this will be further evaluated.

Qualitative data gives an insight and provides useful information that cannot be measured with numbers.

This image depicts a beaker containing water on a hot plate. There is a thermometer

inside to measure 40°C. The temperature reading on there hot plate shows 530°C. This

is only to heat the plate initially before measuring the actual temperature of 50°C.

!4
Sanya Daryani 10B Enzyme Activity Lab April 26, 2019

Processed Data Graph:

Figure 4 - Graph representing the effect of increasing temperature of catalase enzyme from mushrooms on

the average rate of decomposition of H2O2

The Effect of Increasing Temperature of Catalase Enzyme from mushrooms on

the Average Rate of Decomposition of the Substrate H2O2.

Line Equation:

y = 0.1188e-0.008x

Analysis (Interpretation of Data):

Figure 4 illustrated above shows the effect of increasing temperature of enzyme catalase from

mushrooms on the average rate of decomposition of the substrate H2O2. Looking at the average rate of

decomposition of the substrate, the catalase enzyme that was submerged into the 10°C water has an average

rate of 0.12 mL/min. For the 20°C, 30°C, and 40°C increment, the average rate is 0.09 mL/min. Lastly, for

50°C increment, the average rate is 0.08 mL/min.

Referring to the graph, the trend line portrays that the data is negatively skewed and that the average

rate of decomposition of the catalase enzyme gradually decreases. Furthermore, this causes the overall trend

to be linear and slope downwards. With this trend, it is clear that when the catalase enzyme has a higher

!5
Sanya Daryani 10B Enzyme Activity Lab April 26, 2019
temperature, the average rate of decomposition which is measured by the time taken to produce 10 mL of

foam decreases. As the x-axis increases, the y-axis decreases, resulting in an inverse relationship. However,

there is an outlier that doesn’t fit the graph. Referring to the graph, the average rate of decomposition for the

10°C increment is 0.12mL/min and this is the outlier. From the data, it is clear that the point where the

average rate of 10°C lies is the optimum temperature. Theoretically, as mentioned above, the optimum

temperature is 37°C. The graph’s trend line should increase from 10°C to 30°C and then reach its peak close

to 40°C. Once it hits this point, the trend line should slope downwards and pass 50°C due to the denaturation

of the enzyme. In the actual graph, the points start off at its highest point which is at 10°C with an average

rate of 0.12, moving to the next temperature which is 20°C with an average rate of 0.09, and staying at the

same rate until 40°C.

Error bars represent the standard deviation of the data which shows the variation of a set of data

values while error bars are used on graphs to indicate the error or uncertainty in a measurement. It also gives

a general idea of how precise a measurement is and how reliable it can be (how far from the reported value

the true value might be). In addition to this, standard deviations that are small cause data points to become

more reliable. Examining the graph (figure 4), the average rate of decomposition of the hydrogen peroxide,

the 10°C catalase that was extracted from the mushroom has an average rate of 0.12mL/min with a standard

deviation of 0.07. The range of the average rate varies between 0.05 to 0.19. This range is found by

individually adding and subtracting 0.07 to and by 0.12mL/min (the average). The 20°C increment has a

standard deviation of 0.03 and causes the actual rate to vary between 0.06 and 0.12. For the 30°C increment,

the standard deviation is 0.02 and the data varies between 0.07 and 0.11. For the 40°C increment, the

standard deviation is 0.03 varying the data between 0.06 to 0.12. The 50°C increment has a standard

deviation of 0.02 and the data varies between 0.06 to 0.10. Generally, error bars try to prove large variability

in the data and shows that the data is not quite reliable if there is an overlap. In figure 4, the first error bar

extends from the bottom all the way to the top, but not touching the edges, spanning almost the entire graph

vertically. The second and fourth error bar are equivalent in size and are quite large. The third and fifth error

bar are also identical and large too, but is also considered an acceptable range. Moreover, all of the error bars

overlap each other and because of this, it is proven that all the data points are not statistically significant. In a

hypothetical situation, because the data varies, the data points could have been plotted anywhere within the

span of the error bars, thus, resulting with a graph that produced vastly different results. To add on to this, the

data points should have created a positive correlation with a trend line that increases upwards, opposite to the

!6
Sanya Daryani 10B Enzyme Activity Lab April 26, 2019

existent graph which illustrates negative correlated values. An example of this would be if the 10°C

increment had an average rate of 0.05 mL/min data point and ended with the 50°C increment with an average

rate of 0.15 mL/min. As a result, the data is very unreliable due to the large spread of data which is caused by

minor flaws conducted throughout the experiment showing inconsistent results as seen in the raw data table

(figure 2). In figure 2, an example of fluctuant data would be when executing the 10°C catalase test, the

hydrogen peroxide decomposed in 8:21 minutes while in trial 2, it took 21:08 minutes. This data is

demonstrated in majority of the trials, causing it to be unreliable as the error bars touch the trend line.

The graph’s equation of the regression line is y = 0.1188e-0.008x. This equation is extremely useful as

it can predict the rate of decomposition at any given temperature. Specifically, ‘x’ acts as the temperature

value whereas ‘y’ represents the rate of decomposition (mL/min). For example if the temperature of catalase

was 60°C, the average rate would be 0.073511 mL/min, which is then rounded to 0.08 mL/min. Looking at

the optimum temperature, at 37°C, the rate of decomposition would lie exactly on the point 0.0883619464

but is rounded to 0.09 mL/min.

Conclusion:

In conclusion, the graph (figure 4) above conveys how increasing the temperature of catalase

enzyme from mushrooms reduces the average rate of decomposition of H2O2. The graph is exponential and

negatively skewed displaying the data points continuously decrease at a non-linear trend. There is no R2

(coefficient of determination) due to the non-linear, exponential graph where y = 0.1188e-0.008x is the

equation of the line of best fit. There is only one outlier present in the data (the 10°C increment) and this

leads to results that are not reliable. The error bars and inconsistencies in the results prove that the data,

through the raw and processed data table (figure 2 and 3), is unreliable. Additionally, the qualitative

observations that have been made are to support the understanding of the results. Also, through visuals and

descriptions, the experiment would be easier to process as these two factors are used as a guide.

Moving on, based on the analysis of the results, it can be firmly stated that the results do not support

the hypothesis. The hypothesis stated that ‘If the temperature of the catalase in the mushrooms which is

submerged into hydrogen peroxide and detergent increase, then the H2O2 will decompose at a faster rate’.

!7
Sanya Daryani 10B Enzyme Activity Lab April 26, 2019
The expected graph should have been exponential and risen steadily until the anticipated optimum

temperature, which is 37°C, then steeply falls as the enzyme denatures. In reality, the graph produced vastly

different values as there a negative correlation appeared. Additionally, while the temperature increased, the

rate of decomposition of the hydrogen peroxide decreased even though it did show an exponential trend line.

In the actual graph (figure 4), 10°C was the optimal temperature. This increment was supposed to have a

much lower data point, shown in the expected graph (figure 1), leading to a polar opposite. Science can

prove that heat accelerates the catalase activity up to a certain point but due to a few mistakes, the complete

opposite was shown. Moving on to the research question, there was enough information collected to answer

it, but due to inefficient reliability and the lack of support for the hypothesis, the question was able to be

100% answered correctly.

Evaluation/Errors:

Random/systematic/ Weakness/source of Possible effect on data Suggested

both error and magna improvement
Synthetic Reading the mass on the The weighing scale Measure the same core
weighing scale. changes its measure a lot multiple times and
of the time. When the record the value that
mushroom in the beaker appears the most often.
was placed on to the
weighing scale, the mass
kept increasing and
decreasing not giving an
exact value.
Synthetic (zero error) Scale not being on 0g This affects the mass of Make sure to set the
when there is nothing on the mushroom when weighing scale to 0g
it. being measured because before measuring the
excess weight can be beaker that contains the
added accidentally. This mushrooms on to it.
causes the data to be
inaccurate.

!8
Sanya Daryani 10B Enzyme Activity Lab April 26, 2019

Random/systematic/ Weakness/source of Possible effect on data Suggested

both error and magna improvement
Both Environmental factors: Their factors cause an Make sure to conduct the
• Humidity error in data precision experiment in a room
• Vibrations because they can affect that is well isolated and
• Oxygen around the the time taken for the controlled.
room reaction between the
• Air pressure catalase and hydrogen
peroxide to occur.
Random Not stirring the If the mushroom is not Make sure to
mushroom when heating/ stirred well while being continuously stir the
cooling it to get its exact heated/cooled, the beaker mushrooms at a pace that
temperature. will not absorb the is not too fast, while
temperatures carefully, making sure that the part
causing there to be of the beaker where there
miscalculations when are mushrooms, also
measuring the time taken being careful as to not let
as the overall water seep into the
temperature would not beaker.
align with the intended
temperature.
Random Measuring the time taken If the time is not The stopwatch kept on
for the rate of measured properly - if it standby and should be
decomposition of is not stopped at the looked at until the
hydrogen peroxide exact time the 10 mL of reaction produces 10 mL
incorrectly. foam was produced, this of foam.
can cause
miscalculations and also
unreliable.

!9
Sanya Daryani 10B Enzyme Activity Lab April 26, 2019

Random/systematic/ Weakness/source of Possible effect on data Suggested

both error and magna improvement
Random Temperature of each The temperature of each Each solution should be
catalase solution. increment was different. taken out at the same
Each solution was kept time before starting the
our for either too long or experiment and have the
too short. One was either same temperature.
hotter or colder. This can
cause an error in
accuracy since the
temperature is a major
factor in measuring the
rate of decomposition.
Higher temperature
causes a faster rate of
decomposition, vice
versa.
Random Reading the thermometer It becomes difficult to Read the temperature
temperature wrong. monitor the the from eye-level and make
temperature of sure to check whether it
mushrooms and affects is not broken
how data would be beforehand.
presented on a graph.
Random Mushroom not being It affects catalase Make sure to properly
weighed to exactly 3g concentration in the take out 3g and look at
due to researcher solution. This affects the weighing scale until
negligence. precision as different it stops at its exact mass.
measurements of
mushrooms means there
is different timing for
DV measurement,
leading to inconsistent
results.

!10
Sanya Daryani 10B Enzyme Activity Lab April 26, 2019

Random/systematic/ Weakness/source of Possible effect on data Suggested

both error and magna improvement
Random Pouring the right amount The magnitude depends Pay more attention while
of mushroom into the on how much of the pouring as clumsiness
graduated cylinder. mushroom did not get can be a huge impact
mixed with the while executing labs.
hydrogen.
Before pouring the
mushroom into the
graduated cylinder, make
sure to drop it in the
centre of the cylinder, as
best as possible, without
letting it touching the
edges.

Extension/Limitations:

Overall, this experiment was not conducted as well as it should have gone if the instructions were

followed carefully. In spite of this, if this lab had to be repeated, something that should have been done

differently would be to perform more trials rather than five. An ideal number would be around seven trials as

it is not too less or too many when carrying out the experiment. By performing multiple trials, it allows the

data to have more accuracy and reliability when measuring and calculating. Another way to refine the

experiment would be to execute it in a controlled room with a stable environment (humidity, vibrations,

oxygen around the room, air pressure [detailed information on this above]) as this minimizes the chances of

undergoing random and systematic errors. thirdly, while performing this experiment, being careful and

observant when measuring the temperature of the catalase, the time taken to produce 10 mL of foam, and

other processes should be kept in mind (they have also been stated above in the errors table). These are a few

ways to improve the enzyme activity lab.

Bibliography:

Bitesize, BBC. “Animal Organisation - Digestion - AQA - Revision 5 - GCSE Combined Science - BBC
Bitesize.” BBC News, BBC, 2019, www.bbc.com/bitesize/guides/z89mk2p/revision/5.

“Enzymes.” Chemistry for Biologists: Enzymes, Nov. 2004, www.rsc.org/Education/Teachers/Resources/cfb/

enzymes.htm.

!11