Objective

To show

Materials

Methodology
1.

Water was prepared at the following temperatures : 5C, 29C (room

temperature ),

37C, 55C and 70C. Prepare the test tubes as the

following:Test tube
Buffer pH10 (mL)
5mM nitrophenol phosphate
( mL )

2.

1
1.0

2
1.0

3
1.0

4
1.0

5
1.0

0.2

0.2

0.2

0.2

0.2

A controlled test tube was prepared by adding 10mL buffer solution,

0.2 mL substrate and 2 mL of water , soak it in the 37C water.

3.

The contents were added to test tube and were put it in the Water as

shown below

( 1 in 5C, 2 in 29C, 3 in 37C, 4 in 55C and 5

in 70C ).
4.

0.2mL of alkaline phosphate was added in the test tube after 5 minutes

except for control

group

based on the times set. It was kept in the

water for precisely 15 minutes.

5.

8mL 0.02 M NaOH was added after 15 minutes quickly based on time

shown above.
6.

Optical reading was took at 410nM.

Enzyme activity of every test tube is determined using the calibration

graph. Graph of

enzyme activity vs. temperature was plotted.

Result
Test tube

Amount of p-nitrophenol
produced

0.054

0.062

0.061

0.067

0.070

0.009