MEMBER

1.Panasaya
2.Nattawadee
3.Somprathana
4.Chawalnrath

Charoenthitiwong
Lotharukpong
Phuangphairoj
Wongdeshanan

Introduction
This lab is about Factors Affecting Enzyme Lab. The learning objectives of this experiment is to
study how pH effect of enzyme activity. Ammonia has high pH which can be detected by using a simple pH
indicator so we use red cabbage indicator.

Hypothesis
Based on what we have learned, we know that all of enzyme have their own optimal pH so we
think that if the pH is too high or to low enzyme will occur slowly and then denature.

RESULT
Procedure 1 result

Procedure 1 answer

Procedure 2 result

Procedure 2 answer

Investigation of pH result
Investigation of pH result answer
Conclusion

Procedure 1 result

Color

Indicator+
dH2O

Indicator+
Baking soda

Indicator+
Critic acid

Purple

Blue

Pink

Procedure 1 answer
Why is it important to have the three reference tubes?
We need to compare the reaction of each sample. In critic acid we
can see the color changing from purple to red. In bases, we will
use baking soda to change the color from purple to blue. In
neutral solutions, we used distilled water and observe no change.

Can you predict what color of the solution will be if the

urease enzyme convert urea to ammonia?
In this procedure the positive result have to be blue because
when urease enzyme was hydrolyze urea to ammonia that change
the pH was increase.

Procedure 2 result

Color

Indicator+
Urea + Soya
suspension

Indicator+
Urea

Indicator+
dH2O+ Soya
suspension

Blue

Purple

Purple

Procedure 2 answer
What are the purpose of procedure2?
We want to know that the enzyme is working or not. The test
tube that contain both Enzyme and substrate will show positive
result which is blue.

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?
Because if enzyme reaction with urea the color will be change but if no
enzyme the color will still normal so mean that the enzyme will react
with urea only. This test tube cannot change to blue.

Investigation of pH result

Tube 1
Tube 2
Tube 3
Tube 4
Tube 5

Before
1.5
3
3.5
5.5
8

After
5.8
6.59
9
10.45
10.67

+
4.3
3.95
5.5
4.95
2.67

Investigation of pH result answer

How does pH affect the urease activity? How do you know?
Urease is an enzyme its one type of protein so when we add a lot of citric acid the pH will be to low so Enzyme
will be denatured , there will be no color change and will keep its pink color (you can see it from the picture that it has
dregs). If the pH is not to low or too high, enzyme will work. As you can see from tube 1,2,4,5, there is no color change that
much.

NEXT

From your results, what pH would you use and avoid if you have to conduct and experiment using urease enzyme and why?

Adjusting the pH to neutral to low basic.

(7-8)

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?

We can smell of ammonia if it has been produced and pH will increase because ammonia is basic.
We can also check the ammonia by place the litmus paper at the tips of test tube the red litmus paper will be
changed into blue if it has an ammonia.

Conclusion

CONCLUSION
According from the experiment, we added equally
indicators into five tubes. Then, added citric solution and sodium
bicarbonate solution in each tube to make five differences pH. In
first and second tube, pH is lower, and they turned into pink
because we added many drops of citric solution. Proteins from
enzyme that were denatured by acid were precipitate in these
two tubes. We added many drops of sodium bicarbonate
solution into fifth and forth tube as well, so their pH is higher and
turned into green. Third tube has the most optimal pH, which
are accurately working with enzyme. Thus, the result strongly
agrees with our prediction that if pH in enzyme were too high or
too low, enzyme would not work efficiency. Therefore, we can
summarize that pH has an effect with enzyme.

THANK YOU