CHEESE LAB P1

Part 1

Tak Maga
10.24.2017
STEM BIOLOGY AND BIOTECHNOLOGY
PURPOSE

The purpose of this part of the lab was to identify the most efficient way to produce cheese.

HYPOTHESIS

If the four agents Chymosin (FPC), Rennin (NCB), Buttermilk, and Water were tested according to the
procedure, then the FPC agent would have the largest effect on cheese production

PROCEDURE

1. Label the four 6ml test tube 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 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

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 agent.

DATA AND OBSERVATIONS

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 Curdling Time Weight of cone Weight of curds
Agent (min) and curds (g) Weight of cone (g) (g) Rate (mg/min)

Buttermilk 1440 1.79 1.11 1.05 2.01

Chymosin
(FPC) 5 2.32 0.92 1.40 280.67

Rennin (NBC) 490 1.47 0.93 0.79 15.87

Water 2160 1.71 1.07 1.2 0.53

Figure 1: The class averages of our results.

Curdling Time Weight of cone Weight of curds

Agent (min) and curds (g) Weight of cone (g) Rate (mg/min)

FPC wet 5 2.86 0.36 2.5 500

FPC dry 0.91 0.5 0.4 80

NCB wet 26 1.28 0.3 0.98 37.69

NCB dry 0.68 0.59 0.09 3.461

Buttermilk wet 1440 2.5 0.36 2.14 1.486

Buttermilk dry 1.23 1.19 0.04 0.027

Water wet 1440 NA NA NA NA

Water dry 1.34 1.17 0.17 0.118

Figure 2: Our Group Data, both before and after drying.

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 1. The FPC agent produced an almost solidified result in the time frame of 5 minutes

2. The NBC took a longer amount of time, and produced not as solidified results

3. The Buttermilk and Water took an extra day to curdle.

4. The amount of curds left over after drying appeared to be much less than what was left before.

5. The curds were firmly attached to the filter paper.

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ANALYSIS

Figure 3: The curdling time in minutes of the four agents tested in our group.

Figure 4: The weight of curds produced by the four agents, both before and after drying in our group.

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 Figure 5: The rate of production of each of the tested agents after drying in our group.

Figure 6: The rate of production of each of the tested agents after drying in our class.

By analyzing the graphs, it can be seen the ranking of the agents in terms of efficiency. The efficiency

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of an agent can be identified in two factors: Time taken to produce results, and the amount of curds
produced at the end of the process. By dividing these two, it is possible to obtain a curdling rate. As
seen in Figures 5 and 6, the agent with the highest rate was clearly the FPC. Therefore, it is possible to
answer our purpose. The most efficient curdling agent is FPC.

With this result, it is possible to check our hypothesis. It was correct to say that FPC was the most
efficient, therefore proving our hypothesis.

The procedure of the lab provided the means to execute this lab, but also included some issues that may
have affected results. First of all, each group member took charge of some individual cheese curdling
agent. This meant that the cheeses could be followed throughout the lab. However, that also meant that
each stage of the lab was performed by a different person. As there was a lack of communication,
different methods were used to execute each step of the procedure (using different papers for weighing
and filtering for example). This may have lead to a flaw in our data.

In order to remedy these issues with the lab, there are several things that could be done. First of all, the
procedure could be rewritten to include more specific. This could mean that a specific filter paper
would be identified. Another improvement would be to clarify whether the wet weight should be
recorded. This was an item that not every group recorded, and therefore was something that could have
been clarified and reviewed.

Having done this lab, and having investigated some of the agents that are used to produce cheese. This
leads to a further investigation. The time taken to incubate the cheese was, in some cases, longer than
what was anticipated. Therefore, it is conceivable that a higher temperature would lead to a higher
production rate.

CONCLUSION

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These data provide conclusive results. The FPC (Chymosin) agent was the most effective out of the
four that were tested. There are two components to efficacy; time and amount produced. The procedure
tested both. The agents were tested in similar samples and incubated. The time was recorded when the
milk curdled, and the sample weighed before and after the resultants were dried. Through this process,
the time taken to curdle the milk and the amount of cheese produced are measured. In the former
category, shown in Figures 1 and 2, it is starkly clear that the FPC agent produced results in 5 minutes,
compared to other agents ranging from 26 minutes to 24 hours. These data provide evidence that the
FPC is the fastest acting agent. The second component to efficacy is the amount of cheese produced. In
Figure 4, it is shown that the FPC produced 2.5g of curds, compared to the closest competitor
(Buttermilk) at 2.14g. This also defines FPC as the agent that produces the most curds in the
circumstances. Combining these two factors yields us with the result that FPC is the most efficient
agent tested. This is backed up with the results in Figures 5 and 6, both of which put FPC as the most
efficient.

CHEESE LAB P2
Part 2

8
 Tak Maga
10.24.2017
STEM BIOLOGY AND BIOTECHNOLOGY

PURPOSE

The purpose of this part of the lab was to find out whether a higher temperature would lead to a higher
cheese production rate.

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HYPOTHESIS

If the cheese were curdled under a higher temperature, than the cheese production rate would be higher
than under normal temperature.

PROCEDURE
1. Label 2 6ml tubes with curdling process and group number
2. Use large pipet to transfer 3 ml of milk to each tube
3. Use small pipet to transfer the entire contents of each FPC tube into each milk vial.
4. Cap tubes and invert them 3 times, transfer one of them into 95 degree (Celsius) water and keep
the other at body temperature for incubation
5. Check for curdling every 30 seconds for the one in the water and every 5 minutes for the one at
body temperature by gently inverting the tube and examining for curds
6. Record the time in minutes when the milk begins to curdle/solidify
7. In a data table similar to table 1, record time in minutes when it began to curdle or solidify
8. Determine amount produced by each treatment
9. Weigh an empty paper cone and record its weight
10. Transfer the entire contents of the tube on top of the paper and put it over a collection vessel.
Let it dry overnight.
11. Weigh the dry curds on the paper. Subtract the weight of the dry cone. Record the weight of the
curds in mg by multiplying the mass in grams by 1000
12. Repeat for both tubes
13. Record rate of curd production (mg/minutes) in a data table and bar graph

DATA AND OBSERVATIONS

Weight of cone and

FPC Curding Time (min) curds (g) Weight of cone Weight of curds (g)

Heated wet 0.5 3.15 0.38 2.77

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NH wet 5 3.08 0.72 2.35

Heated dry 0.5 1.13 0.38 0.75

NH dry 5 1.47 0.72 0.75

Figure 7: The results of heated and unheated FPC curdling

1. The water began to bubble for a considerable amount of time before we inserted our sample.
2. The control sample was fully curdled when first checked.
3. The higher temperature sample was curdling when first checked, and was removed from heat.
4. Curdling may have continued after removal from heat, as the cheese was almost fully curdled
when being weighed for the higher temperature sample.

ANALYSIS

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 Figure 8: The graph of the results of heated and unheated FPC curdling

From these data, it is possible to first and foremost confirm our hypothesis. As seen in Figure 8, the
time taken to curdle the curds was much less when an increased temperature was applied. After drying,
the amount of curds produced was the same, as seen in Figure 7 (0.75g). From this, we can establish
that cheese curdling occurring under a higher temperature results in a higher rate of production. This
matches with our hypothesis.

There are several errors in the lab that may or may not have occurred. The first was that the temperature
of the water bath may have varied, or was not at the appropriate temperature for the time it took the
cheese to curdle. This was as a thermometer was not placed into the water during the run of the lab.
This, in all probability, did not affect the lab, due to the short amount of time the tube of milk and FPC
was placed in the water bath.

The second possible error may have occurred when we wrote the procedure. We had hypothesized that
the heated milk would curdle at a much higher rate. While this turned out to be true, we adapted the
time at which we checked for curdling to our hypothesis. This meant that we checked the vial in the
water bath at an interval of 30 seconds, compared to 5 minutes with the control. This meant that the
time for the vial in the bath was much more accurate than the control. This may have skewed the data,
but that is not very likely.

In order to improve the lab, it is possible to fix these two specific errors. For the former, it is a simple

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fix. By placing a thermometer within the water bath, we could have obtained an accurate reading of the
temperature. This would have fixed the problem. For the latter problem, by having both readings at an
interval of 30 seconds would remedy that problem.

After concluding this lab, there are other avenues available for research. These mostly involve other
ways to affect production. Some options that are more obvious include a higher temperature and adding
more Chymosin to a similar mixture of milk. All of these would be relatively easy to test: in fact, other
groups in the class experimented with such methods.

CONCLUSION

These data provide a conclusive result. As the temperature for the incubation increased, the time rate of
production of the cheese also increases. This was the result of a procedure that started with two
identical samples of milk laced with the FPC agent. Then, the vials were incubated; one in normal
conditions, and one in a boiling water bath. As seen in Figure 8, the curdling time for the heated sample
was much less than the time for the control. What this means is that the first factor of efficiency (being
mass produced and time taken) is fulfilled. The second factor of efficiency is the mass produced. As
seen in Figure 7, the amount of cheese is the same. This means that the two processes both produced the
same result. However, the time taken by the heated vial was less, and therefore was more efficient. Both
these factors, time and amount produced, are favoring the efficacy of the higher temperature process.
From this, the conclusion drawn is that incubation under a higher temperature will produce cheese more
efficiently.

CHEESE LAB P3
Part 3

13
 Tak Maga
10.24.2017
STEM BIOLOGY AND BIOTECHNOLOGY

PURPOSE

The purpose of this part of the lab was to identify the different macromolecules that were in our cheese.

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HYPOTHESIS

If our cheese sample was tested for four macromolecules (glucose, starch, protein, and lipid), then
glucose, protein, and lipid molecules would be found in the sample.

PROCEDURE

1. Monosaccharide/Glucose
a. Obtain a vial, and place into it a cheese sample that is of the approximate volume of 5
mL.
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 1.5 mL 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 AND OBSERVATIONS

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 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 bag Red Pink Clear

test

Figure 9: Description of positive and negative controls as found by our class

1. Monosaccharide
a. The color changed from blue to a green color.
2. Polysaccharide
a. The color of the cheese did not react in a standard pattern for all groups in the class.
b. A portion of the class found that the color went to a very dark brown/black color.
c. The remainder of the class found a more reddish color.
3. Protein
a. The color of the vial changed from a violet color to a purple color.
4. Fat
a. The color of the cheese using the Sudan IV test was orange
b. The color of the melted cheese was more translucent.

ANALYSIS

MACROMOLECULES FOUND (YES OR NO)

Monosaccharides YES

Polysaccharides/Starch SOME FOUND

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 Protein YES

Lipids YES

Figure 10: Interpretation of the results of the procedure.

What these results mean is not conclusive in its entirety. Using data from control tests, it was possible
to attempt to interpret each of the tests in terms of their results. As seen in Figure 9, four
macromolecules were tested. For the first item, the color of the vial turned a cloudy green. This was
evidence of a reaction, and therefore indicated the presence of monosaccharides in the cheese sample.

For the second item, glucose, the data was not quite as clear. The groups in the class doing the lab did
not produce results that matched each other. Therefore, no conclusion can be drawn from those results.

The third item, protein, was conclusive. The fact that the vial turned a different color was indicative to
the presence of that macromolecule. This result was similar across all tests done both in our lab group
and class.

The fourth item in which we tested the cheese was for the presence of lipids (fats). This was done using
two methods. The first was done using the Sudan IV test. This was another standard color change test.
In this, our class found that there was fat in our sample. This was also corroborated by our second test,
in which the translucency of melted cheese on a piece of paper was tested. In this, we found that the
paper was more translucent, indicating that there was fat in the cheese.

In terms of the hypothesis, it was proved to be accurate in predicting the presence of all three items
(glucose, protein, and fat), and the data was unclear for the remaining macromolecule (starch), which
was predicted to be absent in the sample.

This lab, in terms of procedure, appears to be fairly straightforward. However, problems arose in the
interpretations of results. Even after during the control tests, it was not extremely evident whether a
certain macromolecule was present in the sample. This was exemplified in the starch test (Figure 9), in
which there were different results in the class. Other calls on the remaining molecules were not made
with absolute certainty.

A specific path to improve the lab is not evident. A possibility is to do the positive and negative
controls along with the sample, in order to provide a specific comparison in which to judge the
categorization of the sample. This, however, could lead to other problems with materials and procedure
that would have to be remedied.

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These tests were limited, as they only tested for the presence of the macromolecule, and not the amount.
Further tests might be to identify the percentages of the components of the sample cheese, and the other
molecules that are a portion of the makeup of the cheese. Such tests are, in all probability, beyond the
reach of our class, and would not lead to much benefit.

CONCLUSION

From these data and analysis, it is possible to come to the conclusion that our sample cheese contained
glucose, protein, and fat. Results were obtained through a procedure that started with finding positive
and negative control results for four macromolecules: glucose, starch, protein, and fat. By testing the
cheese sample, it was compared to the controls to identify the components of our cheese. The first result
was that of glucose. The color changed when tested, aligning with our positive control. This positively
identifies it as a component in the cheese (Figure 10). For the second set of results, the positive control
was very similar to the sample when tested for protein. This means that protein was present in our
sample (Figure 10). For the last macromolecule, both the paper test and the Sudan IV test found that the
results corroborated with the positive control. This indicates that lipids (fats) were present in our cheese
sample.

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