TITLE: The Impact of the Enzymes Pectinase and Cellulase on Apple Juice Production
from Apple Sauce
PROBLEM: What is the effect of the enzymes Pectinase, Cellulase or a combination of the
two on the production of apple juice from apple sauce stirred for 8 minutes after 10 minutes
of filtering through filter paper?
VARIABLES:
Independent: the type of enzyme used (Pectinase, Cellulase or a combination of the two)
Dependent: the amount of apple juice produced (mL)
Control: a sample of just apple sauce with no enzyme that is stirred and filtered in the same
way as all other trials
Constants: amount of apple sauce, number of drops of enzyme used per trial, the time
stirred, the method of stirring, the time filtered, the number of drops of water used on the
filter paper,
HYPOTHESIS: If Cellulase (independent variable) is used to produce apple juice from
apple sauce then more apple juice (dependent variable) will be released because Cellulase is
more expensive and therefore this enzyme should speed the reaction more (reasoning).
MATERIALS:
apple sauce
Pectinase enzyme
Cellulase enzyme
funnels
filter paper
lab aprons
splash-proof goggles
paper towels for cleanup
test tube rack
droppers
stirring rods
graduated cylinder
access to tap water and DI water
plastic cups
access to a clock with a second
access to a balance
blue scoops and spoons
test tubes
PROCEDURES:
1) Gather all materials listed above and follow appropriated safety measures.
2) Set up a test tube rack with four test tubes and four funnels placed in the top.
3) Fold four pieces filter paper twice in half and then create a cone with no holes. Place
this paper in each of the funnels and moisten with 15 drops of DI water.
4) Shake the container of apple sauce to ensure uniform mixture before opening. Then,
open the 113 gram container of apple sauce.
5) Using the blue scoop, take one full scoop out of the cup and put it into one of the
plastic stirring cups. To ensure a full cup, overfill the scoop and then scrap the top
of the top of the scoop with the straight end of a spoon.
6) Tape four plastic stirring cups down to the lab table.
7) Carefully remove all the apple sauce from the blue scoop (using the spoon if needed)
and put it into one of the plastic stirring cups.
8) Repeat steps 5-7 until four cups have equal volumes of apple sauce (and a mass of
roughly 28 grams).
9) Label each cup: one control, one Pectinase, one Cellulase and one both enzymes.
10) Into the Pectinase cup drop four drops of room temperature Pectinase enzyme.
11) Into the Cellulase cup drop four drops of room temperature Cellulase enzyme.
12) Into the both enzymes cup drop two drops of Pectinase and two drops of Cellulase
enzyme.
13) Into the control enzyme do not drop any enzyme.
14) Using a different stirring rod stir in each hand stir the two cups clockwise and have a
partner do the same.
15) After 8 minutes have elapsed stop stirring and remove any sauce on the stirring rods
by scrapping it back into the appropriate stirring cup.
16) Label each funnel: one control, one Pectinase, one Cellulase and one both enzymes.
17) Pour the entire contents of the appropriate stirring cup into the correspondingly
labeled funnel with filter paper.
18) Allow the apple juice to drain through the filter for 10 minutes.
19) At the end of 10 minutes remove the funnel from the test tube and then pour the
contents of each test tube individually into a clean graduated cylinder and record the
volume (in mL) of apple juice to the correct precision.
20) Record data in a data table.
RESULTS: The main finding of this laboratory investigation was that 8 minutes of stirring
the enzyme Pectinase in with apple sauce produced a higher average volume of apple juice
after 10 minutes of filtering than any of the other trials.
DATA TABLE:
A Data Table of the Type of Enzyme (Pectinase, Cellulase or Both) vs.
the Volume of Apple Juice Produced (mL) from Stirring with Apple Sauce with Cost Analysis Information
Expense
Volume of
(cents) Per
Apple Juice
mL of Apple
Cost of
Expense
Produced
Juice
Enzyme
Per Trial
(mL)
Enzyme Type
Drops of Enzyme
(cents/mL)
(cents)
Produced
Control
2.1
0
0
0
0.00
Pectinase
7.4
4
5
1
0.14
Cellulase
5.2
4
10
2
0.38
Both
6.1 2 Pectinase & 2 Cellulase
7.5
1.5
0.25
GRAPH:
Control
.
Pectinase
Cellulase
Pectinase &
Cellulase
ANALYSIS:
a. The relationship between type of enzyme and the amount of apple juice produced
after stirring and filtering was that Pectinase performed better than the control, the
Cellulase trial and the Pectinase/ Cellulase combination trial.
b. Enzymes are proteins that act as catalysts. For this particular enzyme catalyzed
reaction there was probably more pectin (the substrate for Pectinase) that was
preventing the release of apple juice from the apple sauce. Therefore, when four
drops of Pectinase was used the most possible juice was released because there were
more available Pectinase enzyme molecules to act on the pectin that was holding up
the release of the apple juice.
c. Since only one trial was conducted due to material and time constraints there wasn’t
the opportunity for outliers.
d. This experiment could have been improved by completing repeat trials for each setup. This experiment also could have been improved by using a clean spatula to
ensure all the apple sauce was removed from the scoop when being transferred into
the stirring cup and also to ensure that all the apple sauce was removed from the
stirring cup when being transferred to the funnel for filtering.
CONCLUSION:
a. What is the effect of the enzymes Pectinase, Cellulase or a combination of the two
on the production of apple juice from apple sauce stirred for 8 minutes after 10
minutes of filtering through filter paper? If Cellulase (independent variable) is used
to produce apple juice from apple sauce then more apple juice (dependent variable)
will be released because Cellulase is more expensive and therefore this enzyme
should speed the reaction more (reasoning).
b. The data support the conclusion that Pectinase has the most effect on the
production of apple juice when stirred with apple sauce. During the Pectinase trial
7.4 mL of apple juice were produced and in each of the other trials less apple juice
was released. Therefore, the original hypothesis was not supported by this data.
Considering cost analysis, Pectinase was the cheaper of the two enzymes used and
produced the most apple juice (with a significant improvement over the control
trial), therefore it makes sense that Pectinase is the most cost effective enzyme
choice, yielding a milliliter of apple juice for 14 cents.
c. The reliability of this data is not full because it was difficult to transfer all of the
apple sauce from the blue scoop to the stirring cup and from the stirring cup to the
filtering funnel. Since some of the apple sauce and presumably some of the enzyme
was lost during transfer the potential amount of juice to be released was decreased
during each transfer. Since the amount of sauce and enzyme lost in each transfer for
each trial is not known the effect of the loss could vary from one trail to the next.
d. The lack of reliability discussed above contributed to an overall lack of validity for
the conclusions from this experiment. Additional trials and better transferring
methods would significantly improve the reliability and subsequently the validity of
the conclusions from this experiment.
e. Future investigations with these conditions could consider the effect of different
amounts of enzyme or different amounts of stirring. The knowledge gained from
this experiment can be utilized in any other situation where food is being produced
and enzymes are being considered to help speed the production of the food product.