Enzyme (Lactase) Inquiry

Group Members names: Talley Kyle, Maegan Sours, Mikayla Dudek

Problem: How does pH affect the enzyme lactase?
Research Hypotheses: (include at least 2 with a prediction)
How does the pH affect the shape of the enzyme and since shape is key to
function, how does that affect how lactose breaks down glucose and
galactose?
We think that the higher the pH the lower the glucose level will be.
We think that Fat-free milk will have better results (Glucose levels) than the
whole milk.
Procedure:
1. set up eight test tubes containing positive and negative controls
testtube #1: glucose (positive)
testtube #2: whole milk (control)
testtube #3: fat free milk (control) ..
testtube #4: lactase (negative) ..
testtube #5: lactase and whole milk (test)..
testtube #6: lactase and fat-free milk (test)..
Have 5 test tubes of #5 and #6:
Testtube 6- Lactase and whole milk
1ml L 1ml M
Testtube 6a- Lactase and whole milk and HCl
1ml L 1ml M .5ml HCl
Testtube 6b- Lactase and whole milk and Vinegar
1ml L 1ml M .5ml Vinegar
Testtube 6c- Lactase and Whole milk and NaOH
1ml L 1ml M .5ml NaOH
Testtube 6d- Lactase and While milk Baking soda

1ml L 1ml M .5ml Baking soda

Testtube 7- Lactase and fat-free milk

1ml L 1ml FM
Testtube 7a- Lactase and fat-free milk and HCl
1ml L 1ml FM .5ml HCl
Testtube 7b- Lactase and fat-free milk and vinegar
1ml L 1ml FM .5ml Vinegar
Testtube 7c- Lactase and fat-free milk and NaOH
1ml L 1ml FM .5ml NaOH
Testtube 7d- Lactase and fat-free milk and baking soda
1ml L 1ml FM .5ml Baking Soda
~add the enzyme before 2 minute timer
2. After 3 minutes test the test tubes pH (mol/L) level with glucose paper. Put
data into Data Table.
Data Table for Testing HCl, Vinegar, NaOH, Baking Soda:
Lactase
and
Whole
Milk
pH
Glucose
Lactase
and
Fatfree
milk
pH
Glucose

Control

HCl

Vinegar

NaOH

Baking
Soda

6.5
150
Control

6.0
200
HCl

7.5
200
Vinegar

8.5
400
NaOH

6.0
0
Baking
Soda

6.7
50

6.0
350

6.1
240

7.3
300

7.8
25 (really
denature
d)

Data Table for Controls:

Glucose

Control

pH
Glucose
Whole Milk
pH
Glucose
Fat Free Milk
pH
Glucose
Lactase
pH
Glucose

6.5
0
Control
6.8
200
Control
6.8
250
Control
6.5
0
https://youtu.be/1gFGU4mTQ9c

-- Whole milk

experiment
https://youtu.be/tT1Oiwo85Gw

experiment

**Control**

-- Fat-free milk

**Fat-free Milk**
Graphs:
https://docs.google.com/a/bvsd.org/spreadsheets/d/1_H8xEdHJ_QDyF
OWFh1Qo-nl8FBI8Q3L9raVbjb4G688/edit?usp=sharing

Analyse Data:
In our lactase and fat-free milk experiment, our prediction was correct.
Our data table displays that the higher pH has lower glucose level. Our pH
was 7.8 and our glucose was 25. This means that the glucose denatured,
explaining why the levels were so low. What was odd to see what a pH of 6.7
and a glucose of 50. What was also odd was seeing the a higher pH have a
lower glucose level than the lower pH. In the lactase and whole milk test, the

highest pH (8.5), had a glucose level of 400. That means that the glucose
was not denatured and seemed to thrive in that environment. Overall in the
fat-free test the acidic situations did better. This is a positive outcome
because the stomach is acidic.

In the lactase and Whole milk solution, the control pH was 6.5. When HCl and
Baking soda was added, the pH level decreased and for the Vinegar and
NaOH, the pH level increased.
In the lactase and Fat-free milk solution, the control was 6.7. When HCl and
Vinegar were added, the pH level decreased but when the NaOH and Baking
soda was added, the pH level increased.
These are interesting results because it looks like the enzyme worked better
in the whole milk solution.
What was expected is that the drastic change in pH would change the shape
of the lactase enzyme making it harder to break down the milk. The
comparative graph helps demonstrate the difference between the pH
environments. What is expected would be the higher pH denatures the
enzyme so the glucose doesn't do very well in that environment.

Conclusion/Recommendations: (Can be bulleted)

Enzyme Conditions: We know that enzymes bond to substances based on
their shape selectivity. Enzymes make reactions occur quickly so the body
can function regularly. In order to do that, there must be a reaction pathway
and low activation energy. Extreme conditions of pH compromise that rapid
ability and therefore, the enzyme cant break down the substance fast
enough when compromised. If the environment is too acidic or too basic, the
enzyme can be inhibited. pH changes the enzyme shape so too acidic or too
basic slows down the reaction. The pH of the stomach is 1.5-3.5. The optimal
conditions for enzymes are less than 5 or higher than 9.
Suboptimal Enzyme Function: Human blood stays around the pH level of
(7.35-7.45). Below or Above means symptoms and disease. If the blood pH is
too high or too low, cells stop functioning and people die. A high fever can
denature, or change shape of an enzyme. That causes less activity for the
enzyme to catalyze reactions in the body. When the environment of the
enzymes are compromised, there are consequences altering the function of
enzymes.
Error: In our experiment in testing the glucose and pH levels affect the
lactase enzyme had too small of a pH range, so next time, we will have to
support a larger range in order to get better results. To get a larger range, we

would but more of the Acid or Base into the solution. Also, there is error in
reading the Reagent Strips. That error could change the perspective of the
environments and change our conclusions.
Biomolecules: Chemistry essentially is the foundation of biology because
everything is made up of subatomic particles. The atomic building blocks and
interactions make biological molecules But are grouped into organic
categories: Carbohydrates, Lipids, Proteins and Nucleic acids. So if
Biomolecules are just larger, organic, groups of molecules, then the same
rules for molecules must apply.
Rates of enzyme activity: The lactase enzyme functions best within
limited ranges of both temperature and pH in its given environment, making
it dependent on both factors for it to perform this essential reaction. Because
the enzyme is dependent on pH and temperature to function efficiently, it is
essential for the environment (in our stomachs and digestive systems) to
have the correct ranges. If lactase is rendered nonfunctional because of
temperature or pH extremes, the breakdown of lactose stops. When the
temperature and pH levels are rendered inefficient, then the result is lactose
intolerance in an organism because the lactase enzyme is not under the
correct environment.
Enzyme Speed: Enzymes are biological catalysts that lower the activation energy for
reactions. By lowering the activation level, the actual reaction can occur more quickly
because it takes less energy to engage the reaction. Reactions more commonly than not
display a physical change, indicating the speed of the reaction, if no physical change occurs,
measuring the pH of a solution after a certain amount of time can indicate how quickly
enzymes have served to speed up the reaction.

Bibliography
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Shmoop Editorial Team. (2008, November 11). Biomolecules and the Chemistry of Life.
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