School of Industrial Technology

Faculty of Applied Sciences

BIO462
BIOCHEMISTRY

EXPERIMENT 3:
FACTORS AFFECTING ENZYMES REACTION

NAME : NAJWA BT AHMAD ABD RAOF

STUDENT’S ID : 2018654492
CLASS : AS2464A
DATE OF EXPERIMENT: 21 /4/2020
Part A: Effect of pH on the activity of amylase in breaking down of starch.

Table 1: Effects of pH on amylase activity

pH Time needed for colour Rate of amylase reaction
change from blue-black to (1/time needed for colour
brown (min) change)
3 11 0.09
5 3 0.33
7 1 2.00
9 15 0.07

a) Draw a graph of pH (x-axis) against rate of amylase reaction (y-axis) with the given data.

pH vs Rate of Amylase Reaction

2.5

2
2
rate of amylase reaction

1.5

0.5 0.33
0.09 0.07
0
0 1 2 3 4 5 6 7 8 9 10
pH

b) Name an enzyme that works optimally at an acidic pH.

- Pepsin

\
 Part B: Effect of substrate concentration on amylase activity

A similar experiment was performed but this time we look at the effect of substrate
concentration on amylase activity, and the results are as tabulated below.

Table 2: Effect of substrate concentration of amylase activity at pH7

Time needed for colour
Amount of substrate (mL) Amount of enzyme (mL) change from blue-black to
brown (min)
1 1 1
5 1 1
10 1 2
15 1 3
20 1 9
25 1 16

a) Draw a graph time needed for colour change (y-axis) against amount of substrate
(x axis) from the data given.

Amount of substrate (mL) vs Time needed for colour

change from blue-black to brown (min)
18
16
16

14

12
Time (min)

10 9

4 3
2
2 1 1

0
0 5 10 15 20 25 30
Amount of substrate (mL)

b) What happened at higher concentration of substrate that caused the rate of reaction
to become steady?
- When substrate concentration increases, the rate of a chemical reaction increases.
However, enzymes become saturated when the substrate concentration is high,
 c) Suggest how can you increase the rate of reaction.
- Increase the temperature of reactant.

Part C: Effect of enzyme concentration on amylase activity

A similar experiment was performed but this we look at the effect of enzyme concentration on
amylase activity, and the results are as tabulated below (Table 3).
Table 3: Effect of enzyme concentration of amylase activity

Time needed for colour Rate of amylase reaction

Amount of enzyme (mL) change from blue-black to (1/time needed for colour
brown (min) change)
0.2 10 0.1
0.4 8 0.13
0.6 6 0.17
0.8 14 0.07
1.0 3 0.33
1.2 3 0.33

a) Draw a graph of amount of enzyme (x-axis) against rate of amylase reaction (y-axis)
from the data given.

Amount of enzyme vs Rate of amylase reaction

0.35 0.33 0.33
Rate of amylase reaction (1/time)

0.3

0.25

0.2 0.17

0.15 0.13
0.1
0.1 0.07

0.05

0
0.2 0.4 0.6 0.8 1 1.2
Amount of enzyme (mL)
b) Can the reaction still happen when no enzyme is added? Explain your answer.
- Yes. The use of enzymes can lower the activation energy of a reaction. However, if
no enzyme added, the rate of reaction will be much slower.