AP Biology: 4th Period

George Padilla (did help with the lab report)

Enzyme Lab:
Lactase reactions

Introduction
During this lab we tested the effects that temperature can have on enzymes. This was an
interesting thing to test because of the heat specificity that enzymes tend to have, if too hot it
can cause denaturation. But, we focused more on what would react with lactase, lactase being
an enzyme has a specific active site, a good analogy of this would be a lock and a key. Without
the proper key the enzyme can not carry out the proper reaction that it was designed for.

Enzymes: What are they?

Enzymes are essential to not only our lives, but lives of plants, protozoans, and more. They
are molecules that act as catalysts and help complex reactions occur, without enzymes some
chemical reactions would take too long, or even not occur at all. What enzymes do to accelerate
or catalyze chemical reactions is that they take molecules called a substrates, and convert it into
molecules called products; and sends its products all over the cell. After finishing this process
they can be reused to do others just the same. However, if the enzyme is exposed to
unfavorable conditions ( temperature, salinity, and pH change) it will cause the enzyme to
become denatured, after becoming denatured the enzyme can not be reused.

Denaturation
Denaturation of an enzyme is the process in which enzymes (and other proteins, as well as
nucleic acids) lose their original quaternary, tertiary, and secondary structure. This process
usually leaves the enzyme as unusable. A enzyme can be denatured by these three things, pH
change, Temperature change, and change in salinity. One or more of these three causes the
deformation of an enzyme, the shape of an enzyme is crucial to its function. If the shape of the
whole enzyme is changed so is the active site, meaning the substrate would no longer fit in its
designated site. No active site for the substrate to go in equals no chemical reaction.

Hypothesis
If lactase was placed in a solution with a pH of 6, and with the temperature of 36C the enzyme
would work perfectly since this environment is favorable for the enzyme. But, if the solution were
to be heated up to 100C then cooled to 5C the enzyme would not work because it denatures
at 53C~60C. Also, lactase would not be able to make glucose from sucrose, because lactase
can only break lactose into glucose and galactose. While sucrose is designed to break down
sucrose into glucose and fructose.

Materials
1.
2.
3.
4.

Test tubes
Lactose (milk)
Test tube racks
Lactase

Neutral pH heated and cooled down (boiled to 100 C for 15 minutes)

Neutral pH solution at room temperature (24

Enzyme in neutral pH solution cooled down to

C)
4C
4. Graduated Cylinders
5. Pipettes
6. Pencil/Pen
7. Paper
8. Glucose strips
9. Timer
10. Tweezers

Procedure
Lactase

Part one: Sucrose and

1. First we tested the sucrose and lactase. Gather five milliliters of both sucrose
and lactase, three tubes of sucrose solutions, and three tubes of lactase solution in total.
After sorting out solutions in different tubes mix each lactase tube with a sucrose tube,
then wait a minute.
2. After waiting a minute get a glucose strip, and use the tweezers to hold the end
of the glucose strip without the colorful box. After getting the right side we dipped each
strip into the mixture and held it for about twenty seconds before removing it.
3. Check to make sure and see whether or not the tip of the glucose strip changed
color, this told us if there was glucose present.
4.

Record data needed from this part of the lab.

Part two: Lactose and Lactase

1. To start this part of the lab we gathered five milliliters of each type of lactase
solution (the three stated in materials*) in three tubes, and five milliliters of milk (lactose)
in three tubes as well.
2. After sorting out the amounts of solutions in each tube we mixed two tubes, one
being a lactase solution, while the other being a lactose solution. After mixing we waited
about a minute to allow for possible interactions to insue.
3. Once mixed we put a glucose strip in the mixture to test for glucose within the
solution. Once in we waited 20 seconds.
4.

Gather data, graph

Data
Sucrose and Lactase:

Change in glucose
levels when added
sucrose

Lactase room
temperature

Lactase chilled
to 4 C

Lactase boiled to
100 C then
cooled

No Change

No change

No change

Lactase room
temperature

Lactase chilled
to 4 C

Lactase boiled to
100 C then
cooled

Change

Change

No change

Lactose and Lactase:

Change in glucose
levels when added
lactose

Control Group:

All lactase solutions

Change in glucose levels when added water

No change

Conclusion
In the lab, we had three different results for all lactase solutions. First, we tested the effect of
temperature on the enzyme lactase. At room temperature lactase reacted with the lactose
solution, we know this because the glucose strips revealed a change in glucose levels.
However, when we tested the heated lactase solution there was no change at all, this is

because of the denaturation of the enzyme. We also added lactase to water as a control group,
of course no glucose was found. Next, we mixed sucrose solutions with lactase solutions, all
three mixtures came back with no glucose changes. This is due to the fact that lactase can only
break down lactose into glucose and galactose. In order for sucrose to be broken down into
glucose the enzyme sucrase is needed which breaks down sucrose into glucose and fructose.
The lab showed the effects of denaturation, and enzyme reaction through the mixture of
different solutions. Our lab gave us real examples of these two things, which allowed me at least
to comprehend it even more. This lab was an interesting one.