Lauren Bertelson

Hannah Gorman
Danielle Honan
Kiera Jost
Enzyme Inquiry Lab

Abstract: The purpose of this lab was to investigate the effect of pH on the functionality of the
enzyme lactase, to create our own experiment and procedure, and to film a commercial for a
product that would improve the functionality of lactase.

Hypothesis: The lactaid will begin to denature at the higher (8,9) and lower (5,6) pHs, and
function best at the ph of the plain whole milk (7.5).

Predictions: If the pH of the solution is not 7.5 the enzyme will begin to denature and lose its
ability to breakdown lactose.

Controls: The same amount of whole milk, and the same amount of lactaid solution is used in
each cup.

Independent Variables: how well the lactaid functions

Dependent Variables: pH of milk

Procedure:
Fill 5 cups with 20 ml of whole milk.
Label the cups 5, 6, 7.5, 8, 9
Add vinegar to cup labeled 6 until the solution reaches a ph of 6.
Add vinegar to the cup labeled 5, and add until it reaches a pH of 5
Add club soda to the cup labeled 8 until the pH reaches 8
Add club soda to the cup labeled 9 until the pH reaches 9.
Add 10 ml of the Lactaid solution to each cup all at the same time and wait 5 min
After the 5 min is up test the solution with the glucose strip and record the glucose content

Results:

Glucose Content
pH (mg/dl)

5 300
 6 250

7.5 100

8 100

9 0

How are rates of enzyme activity in cells affected by pH?

The results from the experiment show that the enzymes begin to denature in solutions
with a higher pH. Lactaid functions best in a solution with a pH of 5. There was a higher amount
of glucose in the solution with a pH of 5 because the enzymes remained intact and did not
denature. When the enzymes are denatured, their shape is changed so that they cannot alter a
particular substance. This is the case with the solution that had a pH of 6, 7.5, 8, and 9. The
enzymes could not hold up the the more basic environment. So, they denatured and were not able
to break lactose into glucose and galactose, resulting in a lower glucose concentration in the
solutions.
We were unable to test the functionality of the enzyme lactase in a pH more acidic than 5,
though we believe that the pH of 5 is about optimal conditions for lactase to work.

Analysis:
1. What products resulted after the hydrolysis of lactose? How do you know this? What is your
proof?

When Lactose is hydrolyzed by the enzyme lactase, it splits into glucose and galactose. In
the experiment if there is a high glucose content that means that a lot of the lactose was broken
down by lactate.

2. Why did you test glucose and the lactase with the glucose test paper?

The resulting solution of the experiment was tested with a glucose strip to measure the
content of glucose. If there is a high amount of glucose that means a high amount of lactose was
broken down into the monomers glucose and galactose. If there was a low amount of glucose
that means only a small amount of lactose was broken down into glucose and galactose.

3. Does untreated milk contain glucose? What is your proof?

 Raw milk contain lactose, and lactose is composed of glucose and galactose. When
lactase hydrolyzes lactose, it produce the monomers glucose and galactose. Because raw milk
contains lactose, it also contains glucose.
 Works Cited

Introduction to Enzymes - Worthington Biochemical. (n.d.). Retrieved November 18,

2016, from http://www.worthington-biochem.com/introbiochem/Enzymes.pdf

Sifferlin, A. (2014, March 10). Study: Raw milk doesn't help lactose intolerance.
Retrieved November 18, 2016, from
http://time.com/17483/study-shows-once-and-for-all-that-raw-milk-doesnt-help-lactose-intoleran
ce/

Siskin, T. J. (n.d.). The Effects of PH and Temperature on Enzyme Activity of Lactase on

Lactose. Retrieved November 18, 2016, from
http://education.seattlepi.com/effects-ph-temperature-enzyme-activity-lactase-lactose-7034.html