Ryan loeber

Stem
11/7/17
Cheese Lab Write Up

Purpose:

The purpose of the first part of this lab was to find out which agent for curdling worked
the most effectively in making cheese out of milk. The four different agents we used were
Chymosin(FPC), Chymosin(NCB), buttermilk, and the bacteria in the milk itself (water). Judging
these methods was basic, all we had to do was base it on how long it took each agent to curdle.
The purpose of the second part of the lab was based on changing a variable from the
first lab and observe the effect the changed variable has on the cheese making process.
The purpose of the third part was to test the cheese we created to see exactly what was
in the cheese. The tests we made would reveal if it contained fat, protein, starch, and sugar.

Hypothesis:

Part 1. If we use FPC, NCB, buttermilk, and water to create cheese from milk, then the
FPC will curdle the quickest.

Part 2. If we add three times as much FPC to create cheese, then it will create more
curds at a faster pace compared to the initial amount.

Part 3. If we test the cheese we made for fat, starch, and sugar then it should reveal that it
contains fats and proteins.

Procedure: Biotechnology: Science for the New Millennium by Ellyn Daugherty

Part 1:
1. Label four 6ml tubes with the type of curdling agent and group number.
2. Use a large pipet to transfer 3ml of milk into 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 tube 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 37 C water bath or
place at body temperature 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 (mins) when the milk begins to curdle.
7. If the milk doesn't curdle in 30 minutes, check for curdling every hour.
8. In a data table similar to Data Table 1, record the time when the milk begins to curdle.
 9. Upon return to the lab, during the next work period, 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 the tube onto a labeled filter paper cone over a collection
vessel. Once all liquid has drained through, dry the filter paper with the curds overnight.
12. Weigh the dry cone with the dry curds. Subtract dry cone weight. Record the weight of
the curds in mg by multiplying the weight in grams x 1,000.
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.
Part 2:
1. Lable two 6ml tubes with the amount of FPC chymosin agent (3x and 1x)
2. Use a large pipet to transfer 3ml of milk into each 6ml tube
3. Use a small pipet to transfer the different amounts of FPC into each labeled tube.
4. Cap and invert the tubes three times and then transfer to 37 C water bath or place at
body temperature.
5. Set a timer and check for curdling every five minutes.
6. Record the time in minutes when the milk begins to curdle.
7. In a data table, record the time in minutes when the milk begins to curdle.
8. Upon return to the lab, during the next work period, determine the amount of curds
produced by each treatment.
9. For each treatment, weigh a paper cone and record the empty cone weight.
10. Transfer the entire contents of the tube onto a labeled filter paper cone over a collection
vessel. Once all liquid has drained through, dry the filter paper with the curds overnight.
11. Weigh the dry cone with the dry curds. Subtract dry cone weight. Record the weight of
the curds in mg by multiplying the weight in grams x 1,000.
12. Repeat with each treatment.
13. Create a data table that reports the rate of curd production (weight/time) by each
curdling agent.
14. Create a bar graph that shows the rate of curd production.
Part 3:
1. Monosaccharide Indicator Test
a. Test for glucose. In a test tube, mix 2 ml of a 2% glucose solution with 2 ml of
Benedicts solution. Heat for 2 minutes in a boiling hot water bath (100 ml of
water in a 250 ml beaker at 100 C) Record all color changes and the length of
time for each color to appear.
b. Test for negative control. In a test tube, mix 2 ml of deionized water with 2 ml of
Benedicts solution. Heat for 2 minutes in a boiling hot water bath (100 ml of
water in a 250 ml beaker at 100 C) Record all color changes and the length of
time for each color to appear.
c. Test for cheese. In a test tube, mix crushed up cheese powder with 2 ml of
Benedicts solution. Heat for 2 minutes in a boiling hot water bath (100 ml of
water in a 250 ml beaker at 100 C) Record all color changes and the length of
time for each color to appear.
2. Starch Indicator Test
 a. Test for starch. In a test tube, mix 2 ml of well mixed starch suspension with 0.25
ml of Lugols iodine. Gently swirl to mix. DO NOT HEAT. Record the color
change.
b. Test for negative control. In a test tube, mix 2 ml of deionized water with 0.25 ml
of Lugols iodine. Gently swirl to mix. DO NOT HEAT. Record the color change.
c. Test for cheese. In a test tube, mix crushed up cheese powder with 0.25 ml of
Lugols iodine. Gently swirl to mix. DO NOT HEAT. Record the color change.
3. Protein Indicator Test (CAUTION: Sodium hydroxide is a strong base, is caustic, and can
burn)
a. Test for protein. In a test tube, mix 2 ml of gelatin (protein) solution with 0.75 ml
of Biuret reagent. Record color change.
b. Test for negative control. In a test tube, mix 2 ml of deionized water with 0.75 ml
of Biuret reagent. Record color change.
c. Test for cheese. In a test tube, mix crushed up cheese powder with 0.75 ml of
Biuret reagent. Record color change.
4. Lipid Indicator Test
a. Test for Lipids. Place a drop of oil (100% fat) on a piece of brown paper bag. Let
it dry for 10 minutes. Hold up paper to light. Record how much light passes
through the spot (% of translucence).
b. Test for negative control. Place a drop of water on a piece of brown paper bag.
Let it dry for 10 minutes. Hold up paper to light. Record how much light passes
through the spot (% of translucence).
c. Test for cheese. Place a drop of water containing suspended cheese powder on
a piece of brown paper bag. Let it dry for 10 minutes. Hold up paper to light.
Record how much light passes through the spot (% of translucence).
d. Test for cheese. Mix 60 l of Sudan IV solution into 2 ml of water containing the
crushed up cheese powder. Red color means negative while an orange color
means a positive for lipids.

Data/Observations:

The data tables below show the results of what came out of the first part of the lab. The first
table contains data about what type of curdling agent were used in the process of creating the
cheese, the amount of time it took (minutes), the weight of the cheese prior to the curds being
separated from the whey, the weight of the dry curds, being the actual cheese, and the rate of
production of the curdling found by dividing the final dry weight by the time it took to create the
cheese. The second table shows similar data but for our second section, which instead of using
 different curdling agents my group used different amounts of the the curdling agents.

The final table below contains the data we found when we tested our cheese for four
components, protein, lipids, starch, and sugar.
 One observation I had was that even through clothing, The test tube and milk was able
to warm up to body temperature. When I took it out of my armpit I was able to feel warmth that
affected some of the agents quicker than others. Another observation I had was the different
smells of each curdling agent once it was fully curdled. Once the FPC Chymosin curdled it had
the worst smell, while the water had to scent and the buttermilk curdles had a better scent. One
other observation I had was the change in color during testing for positive and negative controls
based on what the cheese is composed of. Some reactions would turn colors various times
while others only switched once or none

Analysis
Part 1: The data collected from part 1 of the lab shows that Chymosin FPC was the quickest to
curdle out of the four agents. The bar graph is a good representation of this because the FPC
bar was considerably larger than the others. This proves the original hypothesis that FPC will
curdle the fastest. An error that may have occurred was the different temperatures that each test
tube was held at, given that the different teammates could have different clothing on etc.
Another error could have been an inaccurate measurement of curdling agent, as too little would
make the process slower than normal and too much would increase the process giving and
unreliable result. This experiment could be expanded to test different curdling agents giving
different results.

Part 2: The data collected in part 2 of the lab suggests that using 3x more of the curdling agents
resulted in a larger amount of cheese and a higher rate of production of the cheese. This proves
the hypothesis for part 2 correct, as more curds were produced. Errors that could have affected
this section of the lab include differences in temperature that the vials were kept at, which would
change the curd rates and amounts. Another error could have been the timing. Timing is a big
part of the equation. This experiment could lead to further investigations on improving the
curdling agent, and the relationship it has to other agents.
Part 3: The table containing the data from part 3s tests show what each test will look like if the
substance you test contains each molecule. It also shows which of the four molecules that the
cheese we created has. This data shows cheese has fats, proteins, and glucose, therefore
proving hypothesis 3. An error that may have been made would be to just use big flakes of
cheese mixed into the indicator making it difficult for the two to interact. This would cause the
cheese to crush and suspend in the water allowing it to mix with the indicator solution. This
experiment leads to further studies of different cheeses containing different molecules.

Conclusion
Overall the main takeaway from this lab is that making cheese is not easy. The
experiment consisted of using various curdling agents to make cheese. After the first experiment
of making cheese we ran two other experiments. The second was an experiment where we
changed a variable from the first experiment, the third was an experiment involving testing the
cheese to see was it is made of. The reason making cheese is hard and extensive is because
some agents take a long time to curdle the milk so it may take over 24 hours to show results. As
seen in the data table, NCB, water, and buttermilk all took over 24 hours to curdle. Given that
information, this is a poor way of making cheese. Further studies this experiment may lead to
would be creating different curdling agents are using different ways of heating the vial.
During the process of making cheese a large amount of the mass is lost, proving the
theory that cheese making is a long tough process. This process may be improved by using
different curdling agents that would act faster. More experiment that could be done based on
this process could be to find out which curdling agent is the most effective and make it in a big
batch.