Date: Tuesday 23rd November, 2021.

Title: The effect of temperature on the activity of amylase.

Aim: To investigate the effect of varying temperatures on the activity of amylase.
Theory: Enzymes are biological catalysts produced by genes and are found in the cytoplasm
of cells. They are chemicals which speed up a reaction since the temperature inside
organisms are too low for this to occur and for life to go on. Several factors affect the rate of
enzyme activity such as temperature, enzymes best work at a higher temperature as energy
increases however after a certain temperature, activity levels drop. pH levels, they also work
best at a higher pH level since this affects the shape of the enzyme however if the pH levels
are too high, the substrates may not fit in the enzyme. The enzyme concentration and
substrate concentration also affect the rate of reaction. The more substrates present, the more
likely they are to collide with enzyme molecules. If there are too much substrates, only
increasing the amount of enzymes will increase reaction rate. The same goes for the enzyme
concentration.
Apparatus/Materials: one large beaker, four test tubes, three small beakers, one ice cube, 4ml
of 1% starch, 4 drops of 10% iodine solution, water, Bunsen burner, a tripod stand, one 5ml
dropper, one 10ml syringe, matches, 4ml of saliva (amylase), one thermometer and a
stopwatch.
Method: 1) A water bath was set up and a thermometer placed inside to know when the
water reached 37°C and 55°C. When it reached 37°C, some water was placed into a small
beaker.
2) One small beaker was filled with water and let like that since it was measured to be 20°C
and the other contained water and an ice cube so that it reached 0°C.
3) 1ml of 1% starch and one drop of 10% iodine solution was put into each test tube.
4) The contents were mixed thoroughly until they turned a dark blue, almost black colour.
The test tubes were each placed into a different water bath.
5) The test tubes were left for ten minutes so that they became the same temperature as the
water baths. The time was noted and 1ml of amylase was placed into each test tube. The
tubes were shaken and placed in the respective water baths.
6) The stopwatch was started to note when the dark blue colour faded completely. When this
occurred, the time was written down to the nearest minute.
7) Observations were recorded.
Results: Calculations: amount starch is about 20mg. Starch digested in each minute= 20/time
taken mg/minute.
TABLE SHOWING THE EFFECT OF TEMPERATURE ON THE RATE OF DIGESTION OF STARCH.

Temperature/ °C Time taken/ minutes Starch digested each minute/

mg/min
0 40 0.5
20 20 1
 37 2 10
55 0.5 40
Discussion: As temperature increases, so does the rate of amylase breaking down the
substrate starch. Amylase works best between 37-55°C, as seen in the results above, the
optimum temperature was 55°C. This means the best temperature, that is, the temperature at
which the reaction takes place most rapidly. Amylase activity is faster at higher temperatures
due to an increase in kinetic energy making the enzyme molecules collide more often with
the substrates. Consequently, amylase activity is slower at lower temperatures due to a
decrease in energy, lessening the chance of the enzyme molecules and substrates colliding
with each other. The reaction which occurred here was hydrolysis as seen by the following
equation:
Amylase + Starch ---water- maltose
Sources of Errors/Precautions: Two sources of error are: loss of heat to the environment,
therefore the water baths may have lowered or increased in temperature and people may have
observed the colour differently, making them stop the timer earlier or later. Two precautions
that can be taken are: have more people observe the experiment and wrapping a thick cloth
around the beaker to prevent heat loss.
Conclusion: Amylase activity best worked in higher temperatures than lower temperatures.
Reflection: This lab helped me gain appreciation for the role of temperature and the role
enzymes play in the digestive system.