Date:

Title: Enzyme Activity

Aim: To investigate the effect of temperature on enzyme activity

Apparatus: Liver, dropper, water, three test tubes, thermometer, hydrogen peroxide, stirring
rod, ruler, stopwatch.

Method

1. Test tubes were labelled as 1%, 2%, 3%, 4% and 5% to correspond to the different
concentrations of hydrogen peroxide.
2. The measuring cylinders were used to measure 10ml of hydrogen peroxide
solution for each test tube.
3. The liver tissue was prepared by chopping it into small pieces.
4. 1-2 grams of liver was added to each test tube.
5. The liver tissue was mixed with hydrogen peroxide solution by gently swirling
the test tubes ensuring even distribution.
6. The timer was started immediately upon mixing.
7. The time it took for the gas foam to reach 5cm in each test tube was observed and
recorded. This indicated the rate of the enzyme-catalysed reaction.
8. The experiment was repeated for each hydrogen peroxide concentration, ensuring
fresh liver was used each time.
9. The rate of reaction was calculated by determining the reciprocal of the time taken
for the gas column to reach 5cm (1/time).
10. A table was prepared, and the data was organized including substrate
concentration and corresponding reaction rates.
11. A graph was created with substrate concentration on the x-axis and reaction rate
of the y-axis.

Results

TABLE SHOWING THE TIME TAKEN FOR BUBLLES TO REACH 10 CM, AT

VARIOUS TEMPERATURES

Water bath Temperature/˚C Time taken for the bubbles

 to reach 10 cm (seconds)
Cold 0 20
Hot 65 No reaction
Room Temperature 27 10

Discussion

Enzymes are proteins that help speed up metabolism, or the chemical reactions in our bodies.
They build some substances and break others down. Enzymes create chemical reactions in the
body. Enzymes include detoxification, muscle building, and breaking down food particles
during digestion. Enzymes actually accelerate the rate of a chemical reaction to support life.
Enzyme activity can be affected by a variety of factors, such as temperature, pH, and
concentration. Raising temperature generally speeds up a reaction, and lowering temperature
slows down a reaction. As the pH value is increased above or decreased below the
optimum. pH the enzyme activity decreases. Increasing enzyme concentration will speed up
the reaction, as long as there is substrate available to bind to. The potato extract is the enzyme
used and its found in the bubbles given off when hydrogen peroxide was added to the water
bath. The substrate used was the hydrogen peroxide. Catalase is important in the liver as it is
the enzyme that breaks down harmful hydrogen peroxide into oxygen and water. The
chemical reaction for catalase and hydrogen peroxide is 2H2O2 2O + O2. Catalase
breaks down two hydrogen peroxide molecules into one molecule of oxygen and two
molecules of water in a two-step reaction. During the reaction between catalase and hydrogen
peroxide, oxygen gas bubbles escape and create foam. When the temperature was cold (at 0
degrees) it took the bubbles 20 seconds to reach 10cm. When the temperature was hot (at 65
degrees) no reaction took place and at room temperature (27 degrees) the bubbles took 10
seconds to reach to 10cm. This is so because enzymes are temperature sensitive so therefore
at certain temperature it stops working while at another it works at its optimum (cold
temperature). At 65 degrees there was no reaction therefore the enzyme denatured and
become inactive. Catalase activity increases as the temperature goes from 0˚C to around
30˚C. Catalase activity decreases as temperature goes from around 30˚C to 100˚C.
Temperature is a measure of the kinetic energy of the molecules in a system. Based on the
graph as the temperature increases, the kinetic energy and thus the number of random
collisions of enzyme with substrate increases per unit time. With the increase in frequency in
the number of collisions, the probability of the enzyme and substrate colliding in the correct
orientation (ie. the substrate fits into the active site) also increases. Hence rate of reaction
increases with temperature increases, up until the optimum temperature is attained. A further
increase in the temperature beyond the optimal temperature leads to disruption of the weak
bonds of the enzyme. The unfolding of the protein due to the disruption of bonds is called
denaturing. During denaturation the precise configuration of the active site is lost. As the
active sites are lost, the catalytic activity and hence rate of reaction decrease.

Precautions

1. Avoid thermometer from touching sides of test tube or the bottom of the test tube.
2. Hold ruler vertical to avoid parallax error.
Limitations

1. The hydrogen peroxide was exposed to light for a period, which may have cause it to
decompose, thus causing a slower reaction.

2. The height of the bubbles may not rise to 10cm.

Sources of error

1. Parallax error when reading the meter rule in vertical position.

2. Reaction time when stopping stopwatch.

Conclusion

It can be concluded that approximately 27˚C is the optimum temperature for enzymes, as it
took the fasted time of 10 seconds to reach optimum temperature. On the other hand, extreme
low temperatures of 0˚C causes enzymes to react slowly, as the bubbles took 20 seconds to
reach the 10 cm height. At extreme high temperatures of 65˚C, there was no bubbles present,
as the enzyme was denatured.