Aim: To investigate enzyme kinetics, using different concentration of the enzyme.
Hypothesis: The assay system used in the lab consists of a filter paper disc coated with the enzyme and the dropped into a papercup of substrate (Hydrogen Peroxide). As the hydrogen breaks down the hydrogen peroxide into hydrogen and oxygen gas, the bubbles of oxygen gas collect underneath the filter and make it rise to the surface of the hydrogen peroxide. The time it takes for the filter to rise is an indication of the rate of the rate of enzyme activity.
Rate of enzyme activity-distance time
Materials:
1. A specific weight enzyme 2. Filter papers 3. 3% Hydrogen Peroxide solution 4. A meter rule 5. A forcep 6. A digital stop watch 7. A 50ml beaker
Procedure: 1. Different concentrations of enzyme used are 0.1g, 0.2g, 0.25g, 0.3g, 0.35g, 0.4g, 0.45g, 0.5g. 2. Take 40ml of 3% Hydrogen Peroxide solution in a beaker. Measure the length of the solution in mm. 3. Using forceps immerse the filter paper disc into the catalase solution of given concentration. 4. Allow the disc to absorb the enzyme solution for 5 seconds, then remove it and drain of the excess enzyme solution by touching the filter paper to the edge of the beaker. 5. Drop the disc into 3% Hydrogen Peroxide solution. 6. The oxygen produced from the breakdown of Hydrogen Peroxide by catalase becomes trapped in the fibres of the disc causing the disc to float to the surface of the solution. 7. The time (t in seconds), from the second the disc touches the solution to the time it reaches the surface is an indirect measure of indirect activity. 8. Once the disc reaches the surface record the time taken in a table in your notebook. 9. Clean the beaker and repeat the procedure 4 more times for the required.