Lactase Enzyme Lab p.

Lactase Enzyme Lab

By Kevin Mulligan

Biology 12
January 23, 2013

Lab partners:
Marcella Mondin,
Erika Berg,
Brennan Hagerty
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Abstract
As learned in the Biology 12 course, each substrate has a corresponding
enzyme that it specifically fits with in order to execute the chemical reaction.
Much akin to how a key fits into a specific lock, this “substrate-enzyme complex”
can be modelled by the “Lock and Key Model” (see Figure 1). Observing the
accuracy of this model/theory, five solutions – with mixes of milk, enzyme
solution, denatured enzyme solution, water, and sugar – were made and tested
with glucose strips. The solutions that caused the testing strips to change color
were recorded as having a positive glucose result; the solutions that yielded no
color change were recorded as a negative glucose result. Holding true to the Lock
and Key Model, only the solution with the enzyme lactase and the milk (lactose)
showed signs of a positive glucose result.

Introduction
Lactose is a disaccharide, or sugar, found in dairy products; the presence of
lactose is strongly associated with milk in particular. Lactose can be broken down
into galactose and glucose in a chemical reaction within the body by the enzyme
lactase. Lactose-intolerant people have an insufficient amount of lactase in their
body and, therefore, cannot break down lactose properly for usage.
Sucrose – also a disaccharide – is similar to lactose; it is broken down into
fructose and glucose; however, lactase cannot break down sucrose due to the
sucrose’s shape not fitting in the enzyme.
In this lab, lactase breaking down lactose into galactose and glucose will be
observed. When an enzyme is denatured by adding heat, the results will also be
observed.

Method
Our group prepared three minor solutions prior to the making of the five
main solutions that were to be tested. The first, an enzyme solution, was made by
dissolving one lactase tablet (Lactaid) in a 250 mL beaker filled with 175 mL of
water. The second minor solution was made of 5 grams of table sugar dissolved in
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100 mL of water. The third minor solution was created by adding 20 mL of the
enzyme solution into a pyrex test tube and boiling it on a hot plate for 30 minutes
in a 250 mL beaker filled with 175 mL of water. After we let the third minor
solution cool to room temperature (the 30 minutes of boiling denatured the
enzyme), we began to make the five main solutions to be tested, each in a pyrex
test tube of its own.
We labelled the five solutions A through E. Solution A contained 2 mL of
skim milk (Foremost Dairies) and 1 mL of the enzyme solution. After two minutes,
we tested the solution for glucose using glucose testing strips (Diastix). Solution B
contained 2 mL of skim milk and 1 mL of water; again, after two minutes, we
tested the solution. Solution C included 2 mL of skim milk and 1 mL of the
denatured enzyme solution, Solution D included 2 mL of the sucrose solution and
1 mL of the enzyme solution, and Solution E included 2 mL of the sucrose solution
and 1 mL of water; each of these sat for the two minutes before we tested them.

Results
Solution A yielded a positive response to the glucose test strips; the test
strip’s indicating square turned green which meant that 6 mmol/L were present in
the solution. Solutions B through E all yielded a negative response; the test strip
did not change color, as no solo glucose could be detected.

Discussion
The breaking down of lactose (and only lactose) was successfully carried
out by the enzyme lactase and observed. A diagram of the lactase and lactose
reaction can be found in Figure 2. As affirmed by the Lock and Key Model, only
lactose and not sucrose could be broken down by lactase. Although there is
glucose present in sucrose, the lactase enzyme did not recognize the shape of the
fructose/glucose substrate and could not break it down further for the glucose to
be tested. Because the lactase successfully broke down lactose into galactose and
glucose, the glucose in Solution A could be tested. When the enzyme solution was
boiled in the procedure, the enzyme was denatured by the excessive heat and
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rendered useless; it could no longer break apart the lactose, because its shape no
long resembled the “lock” for the lactose to fit into.
Lowering the pH of the solution surround the enzyme can indeed affect the
enzyme; however, the change must be semi-drastic. For example, the enzyme
lactase is found in an environment within our bodies that has a higher pH than
that of the environment that is the stomach. This is because each enzyme has an
optimal pH; this is the pH that allows the enzyme to work at its full capacity.
Moving lactase to the stomach does alter the pH, but the pH of the stomach acid
is close enough to lactase’s optimal pH that the enzyme can still function. If
lactase was placed in an environment with a pH lower than the stomach- well,
that is quite the change in pH. This could possibly render the enzyme useless, and
it would not be able to function in this less-than-optimal pH.
The reaction in this lab is a hydrolysis reaction. A hydrolysis reaction
involves breaking down a larger molecule into smaller ones; in this lab, lactose
(the large molecule) was broken down into glucose and galactose (smaller
molecules). This can be further edified by the diagram of the lactose and lactase
reaction, again, located in Figure 2.

Appendices

Figure 1. “The Lock and Key Model”

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Figure 2. “The Lactase and Lactose Diagram”