ENZYMES AND FACTORS THAT AFFECT ENZYME

ACTIVITY
Theory
 Enzymes are complex organic substances
produced by an organism to alter the rate of
biochemical reactions that take place in the
cell.
 Show high degree of specificity
 All enzymes have a protein component and are
affected by a number of physical and chemical
factors that denature proteins
- Extremes of pH and temperature
- Presence of metal and non-metal ions
- Presence of alkaloidal reagents
- Enzyme and substrate concentrations
B. Protein Nature of Enzymes
Materials  To 5 mL of the catalase extract, add 5 mL
 1 pc small potato with peel of 3 M NaOH
 Filter paper  Mix the two solutions by tapping the tube
 6% H2O2 against the palm of your hand.
 Add five drops of CuSO 4 solution to the
 Filter paper or cotton/gauze
mixture
 Guaiac solution
PURPLE COLOR
 Saliva FACTORS THAT AFFECT ENZYME ACTIVITY
 Starch solution A. Preparation of Enzyme
 Acidified iodine solution (0.001N)  Extract 10 15mL of saliva from a student donor
 0.1N NaCl and use it to test for the effects of pH and
 0.05N HCl temperature
 0.05N NaOH AMYLASE
 Blender B. Effect of pH
 Funnel Label three test tubes and mix the substances
 Cheesecloth indicated below:
 Wooden splint

Procedures
NATURE OF ENZYME AND ITS EXTRACTION
A. Preparation of Catalase Enzyme
 Grate a potato, including the peel, into a fine
pulp (a blender may be used for this) Add 50
mL of water in a beaker and allow the mixture
to stand for 10 minutes, swirling occasionally
 Strain through cheesecloth and discard the
residue Filter again through filter paper (NOTE:
If filtering through filter paper takes too long,
use cotton or gauze) Use the filtrate for the
following tests
 Mix 5 mL of the filtrate and 5 mL of 6 H 2 O 2
in a test tube Immediately insert a glowing
wooden splint
 Mix well by shaking each test tube and place
in a water bath maintained at body
temperature (37C).
 Recording the time at 3 minute intervals, test
for the presence of starch using 0.001N iodine
solution.
 Record the time needed for the blue color of
starch with iodine to fail to appear.
C. Effect of Temperature
 Label three test tubes, add 10mL of 1% starch
suspension and 1mL of 0.1N NaCl solution.

BIOCHEMISTRY FOR MEDLAB SCIENCE 1

 ENZYMES

 Immerse tube in a beaker of water at room

temperature; tube 2 in a water bath at 37C;
and tube 3 in a boiling water bath.
 Using a pipette, add exactly 2mL of saliva into
each tube. Mix well and monitor in a manner
similar to part II B. Note the length of time it
takes before the blue color with iodine no
longer appears.

BIOCHEMISTRY FOR MEDLAB SCIENCE 2