CHEESE LAB REPORT

Kelsey McIvor
November 1, 2017
Matthews STEM 1

PURPOSE
This lab was conducted to find the best way to make cheese after mixing milk with
agents of chymosin (FPC), rennin (NCB), buttermilk and water.

HYPOTHESIS
Part One: If chymosin (FPC) is mixed with three milliliters of milk in a six milliliter tube,
then the process of making cheese will be the fastest and most efficient.

Part Two: If the amount of milk is increased from three to five milliliters, then chymosin
(FPC) will still be the best agent for making cheese in terms of time and efficiency.

Part Three: If the cheese produced by the lab is made correctly, then the macromolecules
of proteins, lipids and glucose are in this newly created food.

PROCEDURE- PART ONE

1. Label four 6ml tubes 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, natural bovine chymosin 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 37C 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
solidified.
7. If the milk has not curdled in 30 minutes, check for curdling every hour.
8. In a data table similar to the Data Table 1, 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

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collection vessel. Once all liquid has drained through, dry the filter paper with curds
overnight.
12. Weight the dry cone with dry curds. Subtract the dry cone weight. Record the weight
of the curds in mg by multiplying the mass in grams by 1000.
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 that shows the Rate of Curd Production (weight/time) by each
Curdling Agent.

PROCEDURE- PART TWO

1. Label four 6ml tubes with the type of curdling agent and group number.
2. Use a large pipet to transfer 5 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, natural bovine chymosin 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 37C 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
solidified.
7. If the milk has not curdled in 30 minutes, check for curdling every hour.
8. In a data table similar to the Data Table 1, 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 curds
overnight.
12. Weight the dry cone with dry curds. Subtract the dry cone weight. Record the weight
of the curds in mg by multiplying the mass in grams by 1000.
13. Repeat with each treatment.

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

PROCEDURE- PART THREE

1. 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. 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.

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DATA

Part One
CURDLING CURDLING WEIGHT (g.) WEIGHT OF WEIGHT OF RATE
AGENT TIME OF CONE & CONE (g.) CURDS (g.) (mg./min)
(min.) CURDS
Chymosin 7 3.06 0.38 2.68 382.85
(FPC)
Chymosin 20 1.38 0.4 0.98 49
(NCB)
Buttermilk 35 1.5 0.38 1.12 32
Water 46 2.49 0.38 2.11 45.87

Figure One: This data table is a representation of our groups results with each curdling
agent, both before and after drying.

CURDLING CURDLING WEIGHT (g.) WEIGHT OF WEIGHT OF RATE

AGENT TIME OF CONE & CONE (g.) CURDS (g.) (mg./min)
(min.) CURDS
Chymosin
(FPC) 5.651128472 2.59 1.1175 1.22625 178.3193202

Chymosin
(NCB) 1237.142857 1.8 0.78 0.8028571429 7.434636905

Buttermilk 1440 1.95125 0.98125 0.83875 0.5859375

Water 1620 2.4325 1.07 1.12 0.7698784722

Figure Two: This data table shows the class averages of all results.

OBSERVATIONS

1. FPC curdled the quickest and most efficiently in terms of rate (mg./min.) and amount
of byproduct, taking seven minutes to almost entirely solidify.

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2. NCB took much longer than FPC to curdle, but was the second quickest to slightly
solidify.

3. Buttermilk and water had results that differed from group to group, but were the two
slowest agents to curdle with more curds than FPC.

ANALYSIS

Class Average Rate (mg./min.) of Agents

Group Results for Curdling Time and Weight of Curds

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DISCUSSION

The data tables and graphs for the original lab of making cheese are analyzed
based off each, individual agent's efficiency in terms of the time the mixture took to
curdle and the amount of byproduct left. Clearly, FPC took the least time to curdle and
left the most byproduct, the other three agents not close in comparison. However, water
had a considerable amount of byproduct after curdling and NCB had a little higher rate
of milligrams per minute than water and buttermilk.

My hypothesis that if chymosin is mixed with three milliliters of milk in a six

milliliter tube, then the process of making cheese will be the fastest and most efficient
was proven to be correct. I believe this is the case due to FPCs purpose within a young
animal to coagulate and curdle milk within the stomach for an easier digestion process
according to several studies on the common use of this specific agent in the process of
curdling cheese.

Errors that may have affected the labs results can be traced back to the lack of
clarity within the procedure. Each member incubated an agent without multiple trials
to rotate the products among the group and may have led to results directly to the
differences among individuals, including body temperature, consistency, and different
interpretations of the directions. One of the group members also accidentally spilt the
tube and all its contents onto the table, losing an entire agent and those particular
results. This forced us to take numbers from another group and may have been
completely different than the curdling process would have been. To improve this lab, I
would recommend each person work on all four agents as opposed to one, as well as
editing the procedure as a class to ensure that all possible misunderstandings would no
longer be an issue. These results do lead me to further investigation on the effect of
temperature on the curdling time, along with other variables that could be manipulated
to change the rate.

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Part Two

DATA

CURDLING CURDLING WEIGHT (g.) WEIGHT OF WEIGHT OF RATE

AGENT TIME OF CONE & CONE (g.) CURDS (g.) (mg./min)
(min.) CURDS
Chymosin 24 4.57 0.44 4.13 172.08
(FPC)
Chymosin 1,440 4.55 0.4 4.15 118
(NCB)
Buttermilk 30 5.1 0.4 4.7 156.67
Water 50 4.05 0.38 3.67 73.4

Data table of results with 5ml of milk instead of 3

OBSERVATIONS

1. FPC still was the most efficient in curdling time with a total of 24 minutes, but was
closely followed by buttermilk at 30 minutes and water at 50 minutes. The first
twenty minutes of incubation led to no progress, but the FPC and NBC suddenly
began to solidify soon after.
2. All four agents took longer to solidify, but left more curds in the tube.
3. NCB had 0.02 grams more than FPC in weight of the curds. All four agents were
extremely close in production of byproduct with less than a gram in difference.

ANALYSIS/DISCUSSION

The change of the amount of milk from three to five milliliters led to a rather
significant change in results. Rather than FPC being the only agent truly efficient, all four
proved to be successful in time and byproduct amount. However, my hypothesis of if the
amount of milk is increased from three to five milliliters, then chymosin (FPC) will still
be the best agent for making cheese in terms of time and efficiency proved to be right in
terms of rate (mg./min.) and overall efficiency due to the agents purpose within animals.

Data table of curdling rate of all four agents

The errors for this procedure were more obvious this time around due to simple
mistakes. For one, the pipet we used at first was faulty and broke while drawing out FPC,
leading to the loss of some product. The errors from our first procedure also shone
through in this new lab with the procedure, variables dependent on the individual in
control of that particular agent, and lack of clarity within the procedure.

To improve this part of the lab, I would again edit the specifics of the procedure
and check each pipet before use to guarantee a successful transition from the test tube.
These simple fixes may completely change the results of this experiment. For further
investigation, I would be interested in continuing to change the amount of milk to find
the optimal value for making cheese.

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Part Three

DATA/OBSERVATIONS
Standard Indicator Description Description
(molecule of Positive of Negative
tested) Control Control
Glucose Benedicts Blue-yellow- Stayed blue
Solution orange
Starch Lugols Dark Red after 20
iodine brown/black seconds
Protein Biuret Dark Turned blue
reagent blue/purple
Fat Sudan IV Red/orange Clear

Observations within data table showing the positive and negative controls

1. Starch, or the polysaccharides, depended on the group, most finding a red color
while some ended with a dark brown.
2. The color of protein varied in shades of purple.

ANALYSIS/DISCUSSION

MACROMOLECULES FOUND (YES OR NO)

Monosaccharides YES

Polysaccharides NO

Protein YES

Lipids YES

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 The tables above show the results of the experiment and the clear results, the
cheese containing three of the four macromolecules as shown when mixed into the
cheese our group made. My hypothesis of if the cheese produced by the lab is made
correctly, then the macromolecules of proteins, lipids and glucose are in this newly
created food proved to be correct. I believe this is the case because of the chemical
elements behind the process of the cheese curdling, the presence of lipids in milk, and
the addition of each agent.
Errors we made during this experiment was the lack of cheese to test on, resulting
in a slightly less amount than required by procedure. Also, the cheese that we did use
was peeled off the filter, leaving remnants of paper within the cheese pieces and
somewhat contaminating the product. To improve this lab, I would recommend access to
higher quality equipment specifically meant for these sorts of experiments, as well as
more time to undergo the process. For further investigation, I would be interested in
researching the other molecular components of the cheese and how each component
interacts within the cheese.

CONCLUSION
CLAIM: From these three experiments, our group was able to reach the conclusion
making cheese takes the quickest time to curdle and creates the most byproduct when
chymosin (FPC) is the agent mixed with three and five milliliters of milk to curdle that
will later contain the macromolecules of lipids, proteins and glucose.
LEAD IN: The first and second part of this lab was spent testing the effects of mixing the
agents of chymosin (FPC), chymosin (NCB), buttermilk and water with milk to find the
most effective product to make cheese with, while changing the amount of milk from
three to five milliliters to test this variable of the lab. The third and final step was
studying the macromolecules within the cheese we made using Benedicts solution,
Lugols iodine, Biuret reagent and Sudan IV.
EVIDENCE: In the first two steps, FPC was found to be the most effective in the rate of
milligrams per minute curdled, but was surpassed by NCB in the second part of the
experiment in terms of how much byproduct was produced by 0.02 grams. Refer to the
tables and graph in the analysis section to see further detail on the labs evidence.
Changing the milk amount from three to five milliliters truly had an effect on all four
agents and increased the efficiency of all products, particularly buttermilk and NCB.
ANALYSIS: In part one, FPC is significantly higher in the rate of milligrams per minute of
the curdling process than all other three agents. However, the four agents were all
relatively close in efficiency with the added two milliliters of milk. The third part of the
project showed that the only macromolecule not in cheese is polysaccharides, the
product having protein, monosaccharides, and lipids.
REPEAT: All evidence has been stated above and in the analysis section of each part of
the lab.

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