ACTIVITY NO.

8
EFFECT OF ENZYME CONCENTRATION ON ENZYME ACTIVITY

Introduction:
Enzymes are highly specialized class of proteins. They catalyze thousands of
chemical reactions with remarkable property of speeding up chemical reactions. They
combine with the reactants to produce a transition state having less energy than the
transition state of uncatalyzed reaction. When reaction products are formed the free
catalyst is regenerated.
The rate of enzyme catalyzed reaction is determined by the concentration of the
enzyme used. The more concentrated the enzyme used, the faster is the reaction.

Objectives: At the end of the activity, the student can:

1. Determine the effect of different concentration of enzyme on reaction

rate
2. Compute for the concentration of enzyme present in each tube.
3. Determine in which concentration reaction rate is fastest and slowest.

Reagents: Saliva, 6 ml 1% starch solution, 6 drops I2 in KI, 6 ml benedict’s reagent.

Materials: 12 Test tube (20 ml), 10 ml graduated cylinder, test tube rack, brush, 250 ml
beaker, tripod, wire gauze, thermometer, alcohol lamp
Procedure:
1. Prepare 6 test tubes and place 1 ml saliva in each.
2. Add 1ml water to tube 1,2 ml to tube 2, 4 ml to tube 3,6 ml to tube 4, 8 ml to tube
5 and 10 ml to tube 6. Mix thoroughly.
3. Transfer 1 ml of each into 6 separate test tubes (discard excess solution) and add 1
ml of 1% starch paste. Mix well and heat in a water bath with temperature
maintained at 400C for 30 minutes.
4. Divide each of the contents of each tube into 2 and test with:
a. Iodine Test – to half of the content of each tube, add 1 drop of iodine in KI
and note the color produced. Compare the intensity of color in each of the 6
tubes.
b. Benedict’s – to 1 ml benedict’s reagent, add 5 drops of the other half of each
tube and heat in a boiling water bath for 3 minutes. Note color of precipitate.
5. Rank each tube according to decreasing reaction rate one (1) being the fastest to
hydrolyze and 6 being the slowest.