Factors Affecting Enzyme Activity

The purpose of this experiment was to identify at which temperature enzyme activity is the highest. Our hypothesis was that enzymes would be the most active at body temperature (around 36.6℃). We also thought that the tubes which do not contain all of the elements given will not have a reaction.
The materials used were as follows: ice container (with the temperature of 4℃); two water containers (with the temperatures of 40℃ and 80℃); seven glass tubes; water; buffer; catechol oxidase (potato extract); catechol (substrate); stopwatch; pipette; disposable pipette tips; marker.
Firstly, we labelled each tube from 1 through 7 with a marker. Next, we added correct substances (which could be water, buffer, catechol oxidase and catechol) to each tube. The elements added and the corresponding amount can be found in the table of results. After putting the substances in, we immediately started the stopwatch. The first four tubes were left at room temperature of about 22℃. The fifth tube was placed in the ice container, the sixth one in the water container of 40℃ and the seventh one – in the water container of 80℃. The colour of each of the tubes’ inside material was examined after 5, 10 and 15 minutes.
Buffer
Catechol oxidase
Catechol
Temperature (℃)
Initial colour
Colour after 5/10/15 min
1
2ml
1ml
-
-
22
Clear
Clear/ Clear/ Clear
2
1ml
1ml
-
1ml
22
Clear
Clear/ Clear/ Clear
3
1ml
1ml
1ml
-
22
Light yellow
Light yellow/ Light yellow/ Light yellow/ Light yellow
4
-
1ml
1ml
1ml
22
Orange
Light orange with a thin orange layer on top/ Light orange with a thin orange layer on top/ Light orange with a darker layer on top
5
-
1ml
1ml
1ml
4
Light yellow
Light orange with an orange layer on top/ Light orange with an orange layer on top/ Light orange with a dark orange layer on top
6
-
1ml
1ml
1ml
40
Light yellow
Light orange with a thick dark orange layer on top/ Light orange with a thick dark orange layer on top / Light orange with a thickened dark orange layer on top
7
-
1ml
1ml
1ml
80
Light...

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