Effect of

pH on
urease

Introduction

Acid is changing the indicator to red and

Base is changing indicator to blue. pH is
effective to every reaction of the enzyme.
Every enzyme have different range of pH that
can have the best reaction. In this
experiment, we will test how pH affects the
urease enzyme. Which is the best range of pH
to make the reaction happened fastest. We
think that the best range of pH to make the
urease enzyme reaction happed is around 7.

Prediction (Pro 3)
The urease enzyme reacts with urea,
the best pH is 7.

Procedure 1

Results
Name of
tubes

Color of
Reddish
cabbage
indicator

Dropping
(2 drops )

Color after
dropping

Tube A

purple

Citric acid

pink

Tube B

purple

NaCO3

blue

Tube C

purple

Distilled water

purple

Results

Questions

Why is it important to have the three

reference tubes?
Answer: Each tube use to drop three difference
substance. Then compare the color of them
after dropping for know the pH of each tube.

Can you predict what color the solution

will be if the urease enzyme convert
urea to ammonia?
Answer: Dark green or blue because ammonia
is base, which has ph > 7.

Procedure 2:

Fill reddish cabbage indicator 3 ml.

in three tube

Soya
suspension
2ml.
distilled
water
2ml.

Urea 2ml.

Results

Results
Name of Color of
Adding
tubes
indicator substrates
(each substrate
= 2 ml.)

Color
when fills
substrate
s

Color after
adding 5
minute

Tube A

purple

Soya suspension
and 10% urea

blue

green blue

Tube B

purple

10% urea

purple

purple

Tube C

purple

Soya suspension
and distilled water

Light
purple

Light purple

Questions

What are the purposes of procedure 2?

Answer: To test that enzyme is react with the
urea, but not react with distilled water.

Why do we need to add an enzyme

into a test tube without urea even
though we can predict that no enzyme
activity will occur?
Answer: To prove that enzyme is not react
with distilled water.

Procedure 3:

4
drops
2
drops

Drops 3
baking
soda

Drop
acetic
acids

8
drops
6
drops

Results
Name of
tube

Adding
citric
acid

Adding
baking
soda

Color after
vortex

pH by pH
paper
strips

3 ml

Blue

2 drops

3 ml

Purple

4 drops

3 ml

Purple pink

8 drops

3 ml

Pink

16 drops

3 ml

Light pink

Results of pH by using
pH paper strips
Name of tube

pH

Adding 2 ml. of urea solution and

2 ml. of soya suspension.
A

4
drops
2
drops

Drops 3
baking
soda

Drop
acetic
acids

8
drops
6
drops

Results when add 2 ml. of urea

solution.

Test pH by Vernier

After adding 2 ml. of urea solution and 2

ml. of soya suspension.
Measurement by Vernier

Name of
tube

color

Green
blue

Blue

Purple

Pink

Red Pink

pH

10.23

7.58

6.44

4.38

4.42

Question
How does pH affect the urease activity? How do
you know?
Ans. pH 7 is the best state for urease to react with urea.
We saw in the test tubes that the tube which had pH
around 7, the color is changing faster than other, when we
left it about 3 min.

From your results, what pH would you use and

avoid if you have to conduct an experiment
using urease enzyme and why?
Ans. I will use pH 7 to conduct the experiment using
urease because pH 7 is the best state to make a reaction.
pH that I will avoid is 1 because it is a very strong acid and
the reaction might happen very slowly.

Question
Because basic pH will also change the color of the
indicator to blue/green which is very similar when
ammonia has been produced by the urease
enzyme, how can we make sure that the color
change is actually due to the urease activity? Is it
possible to test this?
Ans. Yes, prove by doing an experiment by put the
urea reacts with the ureas first. Then fill the indicator
to see the change.

Conclusion
On this experiment, we have learned about the effect of pH to
enzyme. In this experiment we figure out what is the best range of
pH of the urease to create reaction. The best range of pH to affects
to reaction is around 7 same as our prediction. We can see from the
result that the first test tube which had pH around 7, the color is
changing more and faster from others while leaving for 3 min. We
also ad errors because we dont know how to use the probe to
measure the pH properly. There were 2 tubes that we fill the acid in
different amount, the one that had more acid should had less pH,
but the probe is show up that tube was acidic more than the
another one. Next time we will fix it.

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