CHEESE LAB

Jacob Richards

10.31.2017
STEM 3/4

PURPOSE

The purpose of the lab was to find the most efficient way of making cheese and study
macromolecules.

HYPOTHESIS

Part 1:

If we curdle the milk with Chymosin (FPC), Rennin (NCB), Buttermilk, and Water, then
the FPC agent should be the most efficient.

Part 2:

If we replace our heat source with boiling water, then we should see an increase in the
speed of curdling.

Part 3:

If we use our cheese samples to test for four macromolecules (starch, protein, glucose,
and lipids), then we should see the presence of glucose, proteins, and lipids.

PROCEDURE

Part 1:

1. Label the four 6ml with the type of curdling agent and group number.
2. Use a large pipet to transfer 3 ml of milk to each of the 6ml tubes.
3. Use a small pipet and transfer the entire contents of the tubes of fermentation
produced chymosin (FPC), natural bovine chymosin (NBC), or buttermilk to the
labeled tube containing the milk. For water, fill the small transfer pipet to the
bottom of the bulb and add to the labeled tube containing the milk. Use a different
pipet for each transfer to avoid cross contamination.
4. Cap the tubes and invert the tubes three times and then transfer to a 37 degree
celsius water bath or place at body temperature (i.e. armpit) for incubation)
5. Set a timer and check for curdling every 5 minutes, by gently inverting the tube
and examining for curds.
6. Record the time (in minutes) when the milk begins to curdle (small or large
lumps) or solidify.
7. If the milk has not curdled in 30 minutes, check for curdling every hour.
8. In a data table, record the time (in minutes) when the milk begins to curdle (small

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 or large lumps) or solidify.
9. Upon return to the lab, during the next work period (next day in most lab classes),
determine the amount of curds produced by each treatment
10. For each treatment, weigh a paper cone and record the empty cone weight.
11. Transfer the entire contents of a tube into a labeled filter paper cone over a
suitable collection vessel. Once all liquid has drained through, dry the filter paper
with the curds overnight.
12. Weight the dry cone with curds, Subtract the dry cone weight. Record the weight
of the curds.
13. Repeat with each treatment.
14. Create a data table that reports the Rate of Curd Production (weight/time) by each
Curdling Agent.
15. Create a bar graph the shows the Rate of Curd Production (weight/time) by each
Curdling agent.

Part 2:

1. Label the FPC with group number and name

2. Use a large pipet to transfer 3 ml of milk to each of the 6ml tubes.
3. Use a small pipet and transfer the entire contents of the tubes of fermentation
produced chymosin (FPC)
4. Cap the tubes and invert the tube three times and then transfer to a 100 degree
Celsius water bath
5. Set a timer and check for curdling every minute, by gently inverting the tube and
examining for curds.
6. Record the time (in minutes) when the milk begins to curdle (small or large
lumps) or solidify.
7. If the milk has not curdled in 30 minutes, check for curdling every hour.
8. In a data table, record the time (in minutes) when the milk begins to curdle (small
or large lumps) or solidify.
9. Upon return to the lab, during the next work period (next day in most lab classes),
determine the amount of curds produced by each treatment
10. Weigh a paper cone and record the empty cone weight.
11. Transfer the entire contents of a tube into a labeled filter paper cone over a
suitable collection vessel. Once all liquid has drained through, dry the filter paper
with the curds overnight.
12. Weight the dry cone with curds, Subtract the dry cone weight. Record the weight

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 of the curds.
13. Create a data table that reports the Rate of Curd Production (weight/time) by each
Curdling Agent.
14. Create a bar graph the shows the Rate of Curd Production (weight/time) by each
Curdling agent.

Part 3:

1. Monosaccharide/Glucose
a. Obtain a vial, and place into it a cheese sample that is of the approximate
volume of 5mL.
b. Into this vial, pipet 5 mL of Benedict's solution. Mix well.
c. Heat for 2 minutes in a boiling hot water bath (100 mL of water in a 250-mL
beaker at 100 degrees celsius)
d. Record all color changes
2. Polysaccharide/Starch
a. In a test tube, mix 5 mL of cheese sample with 0.625 mL of Lugols iodine.
b. Gently swirl to mix. Do not heat.
c. Record all color changes.
3. Protein
a. Place 4 mL of a cheese sample in a test tube.
b. Add 1.5mL of Biuret reagent to the test tube.
c. Mix well.
d. Record the color change after 30 seconds.
4. Lipid
a. Paper Test
i. Melt the cheese in a test tube by inserting the vial in a heated water
bath.
ii. Pour the melted cheese onto a piece of paper.
iii. After waiting for the cheese to dry and disperse, hold the paper to
light.
iv. Record the percentage of translucence.
b. Sudan IV Test
i. Add 120 microliters of Sudan IV solution to a 4 mL cheese sample.
ii. Gently mix.

DATA / OBSERVATIONS

Part 1:

Below are the averages of the entire class:

Curdling Time Weight of cone Weight of cone Weight of curds

Agent Rate (mg/min)
(min) and curds (g) (g) (g)

Chymosin
5.65 2.59 1.18 1.23 178.32
(FPC)

Rennin
1237.14 1.8 0.78 0.8 7.43
(NCB)

Buttermilk 1440 1.95 0.98 0.84 0.95

Water 1620 2.43 1.07 1.12 0.77

*Values rounded to the hundredths

Observations:

1. The FPC agent solidified within about 6 minutes for other groups, and 13 minutes
for ours
2. NCB, buttermilk, and water took an extra day to curdle
3. The curds had a white coloration and smelled like a lighter sour milk
4. The volume of curds decreased after drying

Part 2:

Below is the results for our 2nd procedure

Curdling Time Weight of cone Weight of cone Weight of curds

Agent Rate (mg/min)
(min) and curds (g) (g) (g)

Chymosin
(FPC)
0.30 4.44 1.13 3.34 11,133.33
(After
Boiling)
*Values rounded to the hundredths

Observations:

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 1. We noticed that placing the tube into the water had caused some of the water to
splash into the tube
2. The tube, once pulled out of water, had a high temperature
3. The consistency of the cheese was very similar to the cheese made in part 1

Part 3:

Standard Indicator Description of Positive Description of Negative

Control Control

Glucose Benedict's Solution Orange by 20s Light blue

Spreading by 45s
Completely Orange
by 60s

Starch Lugols Iodine Dark brown Light/orange brown

Protein Biuret Reagent Dark blue Light blue

Fat Sudan IV or paper Red Pink Clear

bag test

Observations:
1. The protein test was dark blue
2. The fat test gave a red-pinkish color
3. The Glucose test gave us an almost identical reaction to that of the positive control
4. The starch returned an orange/brown color

ANALYSIS

Part 1:

This graph shows how much more efficient FPC is as a curdling agent

As mentioned in the caption above, and showed by our data, FPC is clearly the fastest
curdling agent.

Our hypothesis is correct, as all of our evidence shows that FPC has an extremely fast
curdling time than the other agents.

There are many errors, but here are 3 examples. The first is the use of unsterile lab
equipment. There were no plugs available to cover our tubes, leading to cheese being
contaminated by our fingers, as we used those for pseudo-plugs. Another error was the
measuring tools that werent accurate. This is mostly because the scales got dirty, wet, or
had other impurities that may have skewed some peoples results, and in turn, skewing

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the average. The last one was quality of direction in terms of measuring mass. The
procedure didnt tell us how to take the mass, making different groups make up methods
that could have impaired their results.

There are 2 ways to improve the lab. The first being getting stoppers for the tubes, so that
milk isnt spilled or contaminated. Another improvement is that everyone should use the
same method of measuring mass so that the results can be as close as possible.

2 investigations arise. One is What is the fastest curdling agent? and the second is Does
heat affect curd production

Part 2:

This bar graph compares the FPC from part 1 to the FPC in part 2

The graph above and the data shows that my hypothesis is correct, in that the rate and
the time taken to curdle have increased after adding more heat to the milk

One source of error was the inability to place and take out the tube quickly, as the water
was boiling hot so it took a few tries to pull out the tube. Another source of error was
human error in terms of tracking time. Its very difficult to pull out to tube and get the
time perfectly, so that may lead to error

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One way to improve the lab would be getting tongs. It's easier to pull out the tube when
you arent burning your hands. If we werent burning our hands, then we could check
the tube more frequently.

This leads me to want to investigate how much can adding heat help, and is there a point
when there is too much heat?

Part 3:

Standard Presence

Glucose YES

Starch NO

Protein YES

Fat YES
Above is a data table showing which macromolecules are present in our cheese

My hypothesis was proven true, as all the macromolecule tests returned positive except
for the starch, which returned negative.

There doesnt appear to be any errors, as there were no major complications in results or
procedure.

Our group had powdered the cheese and mixed it with water in order to test it. This lab
would have been easier and faster if we had a mortar and pestle, or some other device
that crushes things in a consistent manner.

This part of the lab doesnt lead to any further investigations that we could produce. The
results we got were definitive and does not arise any questioning of those results.

CONCLUSION

Cheese contains glucose, protein, and fat and is made fastest when combined with
100 L of FPC per 3 mL of milk in a 100 degree celsius water bath. This experiment was
split into 3 parts. Part 1 was an experiment set to see if FPC was the fastest curdling
agent. We did this by taking 3 other curdling agents, combining all 4 with 100L of their
appropriate agent, heating up these agents, waited for them to curdle, dried the curds,
and recorded the mass. Part 2 was an adaptation of Part 1, where we just took that

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experiment and modified a variable to see the result. We asked the question How does
increasing heat change FPC curd production. We followed similar steps to Part 1, (See
Part 2 Procedure for more info) but mainly focused on changing the incubation method
to a 100 degree celsius water bath. Part 3 focused on testing to see which
macromolecules were in cheese. We first did negative and positive controls on our
solutions. We then powdered our cheese and dumped it into water, which we then tested
on all our solutions.

As shown above, we can see that FPC is consistently the fastest at curdling cheese.
This is due to the fact that FPC is genetically engineered to curdle faster than most agents,
so these results were predictable. This data also shows that boiling the FPC will increase
the number of curds you get

Standard Presence

Glucose YES

Starch NO

Protein YES

Fat YES

Cheese contains glucose, protein, and fat. This was showed by the positive results
we got on all our test except starch, were we got a negative outcome. This is due to the
chemical makeup of curds that stops it from forming starch.

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