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The document summarizes an experiment on factors that affect enzyme activity using salivary amylase. Tests were conducted on the effect of pH and temperature on amylase activity. For the pH test, starch solutions with varying pH levels were incubated and their starch hydrolysis measured over time. Amylase activity was highest within the optimum pH range of 5.6-6.9. The temperature test showed increasing temperature from room to body temperature increased amylase activity, while higher temperatures at boiling point decreased it. In summary, the key factors affecting enzyme activity are temperature, pH, enzyme and substrate concentrations.

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Danica Millesca
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4K views4 pages

Written Report

The document summarizes an experiment on factors that affect enzyme activity using salivary amylase. Tests were conducted on the effect of pH and temperature on amylase activity. For the pH test, starch solutions with varying pH levels were incubated and their starch hydrolysis measured over time. Amylase activity was highest within the optimum pH range of 5.6-6.9. The temperature test showed increasing temperature from room to body temperature increased amylase activity, while higher temperatures at boiling point decreased it. In summary, the key factors affecting enzyme activity are temperature, pH, enzyme and substrate concentrations.

Uploaded by

Danica Millesca
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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PARTIDO STATE UNIVERSITY

College of Education
A/Y 2016- 2017
_____________________________________________________________________________________
_____________________________

EXPIREMENT 4
CHEMISTRY103
GROUP 5
Millesca, Danica Lynn L.
Bermudo, Roselyn B.
Caudilla, Airene P.
Pasa, Emone Rose B.
Cresencio, Siovy S.
Abundabar, Lyka Marie V.

Mix 5 ml of the filtrate and 5 mL of 6% H202 in a test tube. Immediately insert a glowing wooden splint.
What gas is indicated present?

The present of Carbon dioxide.

B. The protein nature of enzymes


To 5 ml of the catalase extract, and add 5 mL of 3M NaOH. Mix the two solutions by tapping the
test tube against the palm of your hand. Add 5 drops of CuSo4 solution to the mixture. Observe and
record the color.

the solution CuSo4 colonized(dark blue)

when adding CuSo4 it becomes lightly coffee brown

there is precipitation

II. FACTORS THAT AFFECT ENZYME ACTIVITY


A. Preparation of enzyme. Extract 10-15 mL of saliva from a student donor and use it to
test for the effect of pH and temperature.
B. Effect of pH
Label 3 test tubes and mix the substances indicated below.
Tube
Number
1
2
3

1% starch
solution
10 ml
10 ml
10 ml

0.1 M NaCl

Acid/Base/Water added

1 ml
1 ml
1 ml

1ml 0.05 N HCI


1 ml distilled water
1 ml 0.05 N NaOH

Saliva
2 ml
2 ml
2 ml

Mix well by shaking each test tube, and place in water bath maintained at body temperature (37C).
Recording the time at 3-minute intervals, test for the presence of starch using 0.001 N iodine solution
.Record the time needed for the blue color of starch with iodine to fail to appear. This means starch has
been completely hydrolyzed to glucose.
Time
(minutes)
3
6
9
12
15

Color of saliva extract with iodine


Test tube #1
-With precipitate
-Has color light brown
-more precipitate
-bubbles appearing
-much precipitate after 9 minutes
-much bubbles
-increasing of precipitate
-increasing of bubbles
-increasing precipitation

Test tube #2
Clear brown

Test tube#3
Lighten (water)

Much clear after 6 mins.

clearer

Much lighten after 6


mins.
Much lighten after 9
mins.
Lighten

Clearer brown

Lighten as water

Much clear after mins.

18

-much bubbles appear


-light brown
-have precipitation
-much bubbles

Clearer brown

Lighten

In what test tube is there evidence of unhydrolyzed starch after 18 minutes?


Observation :

Test tube #1 and test #2


There evidence of unhydrolyzed starch after 18 minutes. In the test #1 and #2 because the starch
has not been completely hydrolyzed to glucose

What is the optimum pH of salivary amalyzed?


The optimum pH of salivary amalyzed is ranges from 5.6 to 6.9.
Conclusion:

We therefore conclude that one of the factors affect the rate at which enzymatic reaction proceed
was the temperature.

C.Effect of the temperature


Label 3 test tube, add 10 mL of 1% starch suspension and add 1 mL of 0.1 N NaCl solution. Immerse tube
#1 in a beaker of water at room temperature , tube#2 in a water bath 37%C,at test tube #3 in a boiling
water bath. Using a pipette, add exactly at 2 mL of saliva into each test tube .Mix well and monitor in a
manner similar to part -B. Note the length of time it takes before the starch is completely hydrolyzed.
Record the time needed before the blue color with iodine no longer appears.
Time(minutes)

Color of saliva extract with iodine


Test tube #1
Test tube #2
Test tube #3
(room temperature)
Water at 37C
(boiling water bath )

Dark brown

Light brown

darker brown

12

-Much darker brown, with


precipitation
-More precipitate than test tube
#2
Darker

darker brown with


precipitation
Much darker brown with
precipitation

15

Darker

Light brown disappearing


of precipitate
Light

18

It looks like brown coffee

Light

QUESTION and ANSWER

-Dark brown at the top


-thick bubbles
-light brown
Lighten brown

-light brown
-less bubbles
After 15mins. There is a
residue at the bottom and its
color is black
After 18mins. It becomes
light brown

1. Write the equation for the reaction catalyzed by catalase.


H2O
2H2O+ O2
2. How does an enzyme alter the rate of a chemical reaction?
The rate of chemical reaction increases as the substrate concentration increases. Enzymes can
greatly speed up the rate of a reaction.
3. What is the optimum pH and temperature for salivary amylase?

Its optimum pH ranges from 5.6 to 6.9 and the optimum temperature of salivary amylase ranges
from 320C to 37oC.

4. What affects do a.)increased pH and b.)increased temperature have on the activity of salivary amylase?
a.)Increased pH
From the optimum conditions, an increase in pH will increase the number of OH- ions, and these
will affect the charge areas on the tertiary structure of the protein (remember that enzymes are
protein). This will cause a conformational (shape) change in the protein (enzyme), and therefore
denatures it, as the active site is no longer complimentary to the substrate. This will lead to fewer
Enzyme- Substrate complexes per second when using a lot of enzymes, and will decreased the
rate of enzyme reaction.
b.)Increased temperature
The rate of an salivary amylase reaction increases as the temperature is raised. A ten degree
Centigrade rise in temperature will increase the activity of most enzymes by 50 100%.
5. In summary, what factors affect enzyme activity?

Several factors affect the rate at which enzymatic reactions proceed- temperature, pH, enzyme
concentration, substrate concentration, and the presence of any inhibitors or activators.

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