Enzyme Lab Report

Enzymes are proteins made up of amino acids used to convert substrates to products. Enzymes
and their active sites are kind of like an interlocking puzzle pieces, only one enzyme fits with its
complementing active site. There are over 2000 known enzymes, and all have a specific
functions. The functions of these enzymes range from speeding up chemical reactions to
breaking down food so that it can be converted to usable energy for a cell. Since enzymes are
substrate specific there is no threat of an unknown reaction or side effect. Enzymes are catalysts.
This means that they make chemical reactions quicker than they would otherwise. In some cases
the essential reactions enabled by enzymes would not happen at all without the help of
enzymes. .Enzymes are not live organisms and do not become part of the final product they
catalyze. They will continue to perform necessary functions and then move on to the next
assignment until it is eventually dissolved by another enzyme.
In order to address the questions about the influences that affect the function of enzymes,
specifically PH, temperature and enzyme concentration we conducted lab investigating how
these factors help or deter the enzyme to break down hydrogen peroxide. The information
obtained will directly correlate with enzymes and how their cellular environment are directly
related in the process of substrate conversions. My hypothesis are based on the following three
variables;
PH Acidic and basic are two extremes that describe a chemical property chemicals. Mixing acids
and bases can cancel out or neutralize their extreme effects. A substance that is neither acidic nor
basic is neutral. The pH scale measures how acidic or basic a substance is. The pH scale ranges
from 0 to 14. A pH of 7 is neutral. A pH less than 7 is acidic. A pH greater than 7 is basic.
Temperature is another environmental variable that enzymes are affected by. We learned that
diffusion is The spontaneous tendency of a substance to move down its concentration gradient
from a more concentrated to a less concentrated area. During diffusion the more heat the faster
the molecules move, and the colder temperatures they are exposed to results in slowing of
movement until it ceases altogether.
Concentrate gradient The concept of osmosis and water concludes that concentrated gradient
conditions are neutralized by water infiltration. Water is a molecule that acts as a neutral party,
anytime there is a high concentration water will move to break up the gradient area.
Hypothesis 1: With the lower Ph , more acidic,
Hypothesis 2: Temperatures ranging from freezing to will result in none to slow enzyme
function. At room temperature or body temperature the enzyme will react at optimum
performance and as temperatures increase there will be no change in enzyme function process.
Hypothesis 3: As enzyme concentration lessens so will the speed at which the enzyme catalyzes.
With higher concentration enzymes will catalyze at optimum levels until it eventually levels out.

Methods:
Effects of pH on enzyme activity
1. Obtain four test tubes and label each test tube. Labels: pH 2, pH 5, pH 7 and pH 12
2. Fill each test tube with 3ml of fresh potato juice enzyme solution
3. Add 2 ml of buffer solution as follows
a. pH 2 buffer solution to the test tube labeled pH 2
b. pH 5 buffer solution to the test tube labeled pH 5
c. pH 7 buffer solution to the test tube labeled pH 7
d. pH 12 buffer solution to the test tube labeled pH 12
4. Mix gently and let sit for 5 minutes
5. At the end of 5 minutes add 1 ml of hydrogen peroxide substrate to each tube
6. Mix gently and let sit for 5 minutes
7. After 5 minutes measure the amount of foam using a millimeter ruler
8. Recorded my results in a data table.

Effects of temperature on enzyme activity

1. Obtain four test tubes and label each test tube. Labels: Ice, Room Temp, 45 C, 70C
2. Fill each test tube with 5ml of fresh potato juice enzyme solution
3. Make and ice bath by placing 50ml of ice in a 100 ml beaker and covering it with water.
4. Make a room temperature bath by adding 50ml of tap water to a 100 ml beaker.
5. Place the labeled test tubes as follows;

a. Test tube labeled ice in the ice bath

b. Test tube labeled room temp in the room temperature water bath
c. Test tube labeled 45 C in the test tube rack in the 45C water bath
d. Test tube labeled 70C in the test tube rack in the 45C water bath
6. Let sit for 5 minutes in the water baths
7. At the end of the 5 minutes leave the labeled test tube in their water baths and add 1 ml of
hydrogen peroxide substrate to each tube.
8. Mix gently, let sit for 5 minutes
9. After 5 minutes measure the amount of foam using a millimeter ruler
10. Recorded my results in a data table.
Effects of substrate concentration on enzyme activity.
1. Obtain 4 test tubes and label each test tube. Labels: 0ml, 1ml, 3ml, and 5ml
2. Fill each test tube with 3ml of fresh potato juice enzyme solution
3. Add hydrogen peroxide substrate to each tube as follows;
a. Test tube labeled 0ml, add 0ml of hydrogen peroxide
b. Test tube labeled 1ml, add 1ml of hydrogen peroxide
c. Test tube labeled 3ml, add 3ml of hydrogen peroxide
d. Test tube labeled 5ml, add 5ml of hydrogen peroxide

4. Mix gently and let sit for 2 minutes

5. After 2 minutes measure the amount of foam using a millimeter ruler
6. Record results in a data table
7. Repeat experiment

Effects of enzyme concentration on enzyme activity

1. Obtain 4 test tubes and label each test tube. Labels: 0ml, 1ml, 3ml, and 5ml
2. Add potato juice enzyme to each tube as follows;
a. Test tube labeled 0ml, add 0ml of potato juice enzyme
b. Test tube labeled 1ml, add 1ml of potato juice enzyme
c. Test tube labeled 3ml, add 3ml potato juice enzyme
d. Test tube labeled 5ml, add 5ml potato juice enzyme
3. Add 3ml of hydrogen peroxide substrate solution to each labeled test tube
4. Mix gently and let sit for 2 minutes
5. After 2minutes measure the amount of foam using a millimeter ruler
6. Record results in a data table
7. Repeat the experiment.
Results.
EFFECTS OF PH ON POTATO JUICE ENZYME ACTIVITY (fig 1)
pH
Group A (ml of
foam produced

pH 2
0

pH 5
0.5

pH 7
4.5

pH 12
3.5

Group B (ml of
1.5
1
2
0.5
foam produced)
AVERAGE
.75
.75
3.25
2
EFFECTS OF TEMPERATURE ON POTATO JUICE ENZYME ACTIVITY(fig 2)
pH
Group A (ml of
foam produced

Ice
0

Room Temp
0.5

45 C
4.5

70 C
3.5

Group B (ml of

1.5

0.5

foam produced)
AVERAGE

.75

.75

3.25

EFFECTS OF SUBSTRATE CONCENTRATION ON POTATO JUICE ENZYME ACTIVITY

(fig 3)
Substrate Amt
ml of foam
produced

0 ml
0

1ml
0.7

3ml
2.5

5ml
2.7

ml of foam
0
4
1.5
11.5
produced
AVERAGE
0
2.35
2
7.1
EFFECTS OF ENZYME CONCENTRATION ON POTATO JUICE ENZYME ACTIVITY
(fig 4)
Substrate Amt
ml of foam
produced

0 ml
0

1ml
5

3ml
12.5

5ml
10.9

ml of foam
produced
AVERAGE

7.5

8.75

9.2

As seen in the charts above (fig.1 and fig 2):

Enzymes perform at optimal levels when in a neutral pH environment or around 45C. The more
acidic the less catalyzing. The more alkaline a solution is the closer to optimal performance The
colder the less movement and higher heat caused more movement. The foam bubbles are used to
measure the oxygen being given off.
(As seen in fig 3 and 4)
The general result shows that the higher the concentration of substrate or enzyme resulted in
more oxygen produced.

Discussion and Conclusion.

From our data that was collected, it can be concluded that the higher the
concentration of potato or substrate, the quicker oxygen is released from the
hydrogen peroxide and potato. This was expected from our hypothesis. This
could be because there is more surface area for the enzymes to work at. We
also know that heat plays a factor in enzymes performance as proven in our
data from the lab. I think the results may have been a bit inaccurate from
group to group because of the difference in times. Perhaps measuring gases
emitted would have proven a bit more accurate.
Enzymes are also proteins that are folded into complex shapes that allow
smaller molecules to fit into them. The place where these substrate
molecules fit is called the active site. If the shape of the enzyme changes, its
active site may no longer work.
Enzymes are the workers of the body. When you eat, enzymes break down
the food into tiny particles which can be converted into energy in the body.
Enzymes are biological catalysts and are necessary for life to occur. This is
due in part to the fact that enzymes can only function in certain pH,
temperature, enzyme concentration and substrate concentration levels
which took part to react with the abiotic factor oxygen which affected the
Catalase enzyme. From labs and research it is known that when levels rise or
fall below these levels, it will affect the enzyme, and in turn the organism. At
a moderate level the enzyme will work properly, and at extreme or high
levels of acidity or heat the enzyme will break down, and the reaction will
never occur

Memorie Brown
Robert Golman
February 13, 2015
Biology 101