Final 6C/D Report 2024
In this investigation, I looked at how temperature affected the activity of the enzyme amylase, which
catalyses the conversion of starch to maltose. The objective was to examine how enzyme activity
varies with temperature and to identify the ideal temperature at which amylase has the maximum
catalytic activity. The spotting tile approach was the method used. The goal of the study was to
determine the temperature at which amylase activity peaks. It also aimed to identify indicators of
denaturation at high temperatures and investigate the effects of low and high temperatures on
enzyme function. Up to 40°C, the ideal temperature, amylase activity rose with temperature. Above
50°C, activity quickly dropped, indicating breakdown-induced denaturation of the enzyme. Enzyme
activity decreases at low temperatures (20°C) because to a reduction in molecular mobility and
reaction rates.
My hypothesis is that amylase activity will rise with temperature until it reaches an ideal level, at
which point denaturation of the enzyme will cause the activity to fall. Amylase activity is not much
impacted by temperature.
The is the first method for amylase at different temperatures that we used , however the
experiment didn't perform.
● Prepare amylase and starch solutions.
● Temperatures of warm water baths include 0°C, 20°C, 40°C, 60°C, and 80°C.
● Put the amylase and starch in a test tube and set a timer.
● Transfer a drop to a spotting tile well filled with iodine every 30 seconds.
● Observing the colour change blue-black indicates the presence of starch, while yellow
indicates the breakdown of starch.
● Take note of the time once the iodine has ceased changing colour.
This is the second method we used for amylase at different temperatures
● Prepare amylase and starch solutions.
● Temperatures of warm water baths include 0°C, 20°C, 40°C, 60°C, and 80°C.
● Keep the amylase and starch in the separate test tubes
● Then add a drop of starch solution to the iodine — the mixture should turn blue-black,
indicating the presence of starch.
● Start the reaction by adding the amylase solution to the starch-iodine mixture.
● Transfer a drop to a spotting tile well filled with iodine every 30 seconds.
● Observe and time the colour change blue-black indicates the presence of starch, while yellow
indicates the breakdown of starch.
● Take note of the time once the iodine has ceased changing colour
The reason behind changing the way we added the starch and amylase is because we made the
previous mistake of mixing both solutions together first , before adding it to the iodine spotting tiles
which meant the reaction started immediately . Therefore, the amylase began breaking down the
starch immediately, so part of the starch may have already been transformed to maltose by the time
the mixture was added to the iodine. This could have prevented any colour shift at all. Accurately
analysing the impact of temperature would be more challenging since it would be more difficult to
, quantify the exact amount of time it takes for the starch to break down because the reaction would
have already begun. The purpose of the iodine test is to demonstrate how starch breaks down over
time. You cannot monitor the response from the start if the amylase has already started to function
before the iodine is introduced, which might compromise the accuracy of my data.
Temperat Time taken Mean Rate or
ure for no further time (s) 1/t (s^-1)
colour change
(s)
Repeat 1 Repeat 2 Repeat 3
20 1858 1869 1909 1878
30 350 328 308 329
40 860 853 847 853
50 170 134 181 162
60 2000+ 2000+ 2000+ 2000+
I will now do my calculations , Null Hypothesis (H₀): The success of starch breakdown is independent
of temperature. I did two statistical tests , the T-test and the chi test.
Expected value calculation:
T-test calculations ( for 20 and 30°C)
● Xˉ1 =1878.67 (mean for 20°C)
● Xˉ2= =328.67 (mean for 30°C)
In this investigation, I looked at how temperature affected the activity of the enzyme amylase, which
catalyses the conversion of starch to maltose. The objective was to examine how enzyme activity
varies with temperature and to identify the ideal temperature at which amylase has the maximum
catalytic activity. The spotting tile approach was the method used. The goal of the study was to
determine the temperature at which amylase activity peaks. It also aimed to identify indicators of
denaturation at high temperatures and investigate the effects of low and high temperatures on
enzyme function. Up to 40°C, the ideal temperature, amylase activity rose with temperature. Above
50°C, activity quickly dropped, indicating breakdown-induced denaturation of the enzyme. Enzyme
activity decreases at low temperatures (20°C) because to a reduction in molecular mobility and
reaction rates.
My hypothesis is that amylase activity will rise with temperature until it reaches an ideal level, at
which point denaturation of the enzyme will cause the activity to fall. Amylase activity is not much
impacted by temperature.
The is the first method for amylase at different temperatures that we used , however the
experiment didn't perform.
● Prepare amylase and starch solutions.
● Temperatures of warm water baths include 0°C, 20°C, 40°C, 60°C, and 80°C.
● Put the amylase and starch in a test tube and set a timer.
● Transfer a drop to a spotting tile well filled with iodine every 30 seconds.
● Observing the colour change blue-black indicates the presence of starch, while yellow
indicates the breakdown of starch.
● Take note of the time once the iodine has ceased changing colour.
This is the second method we used for amylase at different temperatures
● Prepare amylase and starch solutions.
● Temperatures of warm water baths include 0°C, 20°C, 40°C, 60°C, and 80°C.
● Keep the amylase and starch in the separate test tubes
● Then add a drop of starch solution to the iodine — the mixture should turn blue-black,
indicating the presence of starch.
● Start the reaction by adding the amylase solution to the starch-iodine mixture.
● Transfer a drop to a spotting tile well filled with iodine every 30 seconds.
● Observe and time the colour change blue-black indicates the presence of starch, while yellow
indicates the breakdown of starch.
● Take note of the time once the iodine has ceased changing colour
The reason behind changing the way we added the starch and amylase is because we made the
previous mistake of mixing both solutions together first , before adding it to the iodine spotting tiles
which meant the reaction started immediately . Therefore, the amylase began breaking down the
starch immediately, so part of the starch may have already been transformed to maltose by the time
the mixture was added to the iodine. This could have prevented any colour shift at all. Accurately
analysing the impact of temperature would be more challenging since it would be more difficult to
, quantify the exact amount of time it takes for the starch to break down because the reaction would
have already begun. The purpose of the iodine test is to demonstrate how starch breaks down over
time. You cannot monitor the response from the start if the amylase has already started to function
before the iodine is introduced, which might compromise the accuracy of my data.
Temperat Time taken Mean Rate or
ure for no further time (s) 1/t (s^-1)
colour change
(s)
Repeat 1 Repeat 2 Repeat 3
20 1858 1869 1909 1878
30 350 328 308 329
40 860 853 847 853
50 170 134 181 162
60 2000+ 2000+ 2000+ 2000+
I will now do my calculations , Null Hypothesis (H₀): The success of starch breakdown is independent
of temperature. I did two statistical tests , the T-test and the chi test.
Expected value calculation:
T-test calculations ( for 20 and 30°C)
● Xˉ1 =1878.67 (mean for 20°C)
● Xˉ2= =328.67 (mean for 30°C)
