CHEESE LAB REPORT

Part I: Cheese Production

Nihal Nazeem

November 1, 2017

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PURPOSE

To test multiple curdling agents: Water, Buttermilk, Renin (NCB), and Chymosin (FPC), to
understand how it affects cheese production.

HYPOTHESIS

If 1 vial of 100L of 1 mg/mL of Chymosin FPC (Fermentation Produced Chymosin) is

used for cheese production, then it will make the most cheese at the fastest rate of
mass/time.

MATERIALS

1. 12 mL of milk
2. 1 vial of 1 mg/mL Fermentation Produced Chymosin (FPC) (100L)
3. 1 vial of 1 mg/mL Natural Chymosin from Bovine (NCB) (100L)
4. 1 vial of 1 mg/mL Buttermilk (100L)
5. 1 vial of water (2mL)
6. 4 pieces of of filter paper
7. 4 empty 6mL tubes
8. 1 3 mL transfer pipet
9. 4 small transfer pipettes

PROCEDURE

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.

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 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 agent.

OBSERVATIONS:

The Milk used was in the open air

Caps were not used to close the tubes, thumbs were used instead.
The students armpits were used for incubation
Approximately 3 mL of milk was added
A dull-sour smell was experienced after the first inversion
Chymosin was completely solidified, but the Renin was semi-solidified with some
whey, as Buttermilk had
Cheesy smell after the curds dried overnight

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DATA

Class Averages:
Curdling Curdling Time Weight of Cone Weight of Cone Weight of
Agent (min) and Curds (g) (g) Curds (g) Rate (mg/min)
Chymosin
(FPC) 5.651128472 2.59 1.1175 1.22625 178.3193202
Renin (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

In-Group Data:
Curdling
Curdling Time Weight of Cone Weight of Weight of Curds Rate
Agent (min) and Curds (g) Cone (g) (g) (mg/min) Comments
Chymosin 229.235880
(FPC) 5:01 3.51 2.36 1.15 4
Renin
(NCB) NA NA NA NA NA Completely Spilt
0.79861111 Inaccurate Timing
Buttermilk 1440 3.53 2.38 1.15 11 and Wet Weight
0.79861111
Water 1440 4.21 3.06 1.15 11 wet weight

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ANALYSIS

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Looking at the first graph above, it is clear that Chymosin (FPC) was the most
efficient curdling agent when comparing the rate of cheese as the amount produced (in
mg) per minute. The way these cheese production methods work are as follows: Milk
may be left to age, exposed to air and naturally occurring bacteria; New batches of
cheese are started with specific cultures of selected bacteria, and these known bacteria
make enzymes that uses milk sugars (lactose) to produce lactic acid causing milk protein
(Casein) to denature (unwind) and fall out of the solution. These lumps of denatured
Casein are curds. Buttermilk, yogurt and other fermented products have a good culture
of Lactobacillus bacteria and can be used as a starter. New cultures can be started by
the addition of purified enzymes (such as Renin, from the cell lining of the stomachs of
cows) to milk. Renin is a type of protease made in nursing calves, which cleave the
Casein protein in milk into small fragments that settle out as curds. Genetic engineering
techniques are used to transfer the cows DNA code for Chymosin into fungus cells.
Fungus cells then read the cow DNA and synthesize the Chymosin enzyme, which
scientists call Fermentation Produced Chymosin (FPC). Cheese-makers can use the
genetically engineered chymosin enzyme to curdle milk. Chymosin (FPC) may have

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produced the most, as it is genetically engineered to produce the most, for a given
amount of time. By the results, the hypothesis that 1 vial of 100L of 1 mg/mL of
Chymosin FPC (Fermentation Produced Chymosin) used for cheese production, will
make the most cheese at the fastest rate of mass/time, is clearly supported, as it had
produced the most cheese per unit time. Some errors that this lab may have encountered
could be the use of thumbs as tube caps, leading to contamination and slight alteration of
the chemical reactions of the solutions. Another error that could skew results could be
attributed to improper sterile technique, where the pipette could have
cross-contaminated other solutions. In addition, the procedure was not clear on weighing
methods, whether to use the wet or dry weight or when to weigh, causing groups to have
different and non-standardized measurements, and the data could be skewed as some
groups also had no data to contribute. Some ways the lab could be improved could be by
setting a class standard on the degree of significant figures for data, using tube caps, so
that the solution is not spilled or contaminated, and a standardized weighing method.
Some further investigations that this leads to are about the effect of types of milk on each
curdling agents rate of cheese production, and the role of different temperatures in
curdling.

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CHEESE LAB REPORT

Part II: Cheese Production Manipulation

Nihal Nazeem

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PURPOSE

Understand the scientific process, to design an experiment that manipulates variables.

HYPOTHESIS

If 1 vial of 200L of 1 mg/mL of Chymosin FPC (Fermentation Produced Chymosin) is

compared to 1 vial of 100L of 1 mg/mL of Chymosin FPC, then the rate of curdling in
mg/min should be higher for the 200L of 1 mg/mL of Chymosin FPC .

MATERIALS

1. 12 mL of milk
2. 1 vial of 1 mg/mL Fermentation Produced Chymosin (FPC) (100L)
3. 1 vial of 1 mg/mL Natural Chymosin from Bovine (NCB) (100L)
4. 1 vial of 1 mg/mL Buttermilk (100L)
5. 1 vial of water (2mL)
6. 4 pieces of of filter paper
7. 4 empty 6mL tubes
8. 1 3 mL transfer pipet
9. 4 small transfer pipettes

PROCEDURE

1. Label the two 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 100 Chymosin
(FPC) and 200 Chymosin (FPC) 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

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 celsius water bath or place at body temperature (i.e. armpit) for incubation)
5. Set a timer and check for curdling every 2 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 dry 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.

OBSERVATIONS:

Faint cheesy smell

More liquid (whey) with some curds for the 100 Chymosin (FPC)
More solidification than 100 Chymosin (FPC), with little whey, for 200 Chymosin
(FPC)

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DATA

Cheese Production Rate Compared to Amount of Chymosin

Curdling Time Weight of Cone Weight of Cone Weight of Curds
Curdling Agent (min) and Curds (g) (g) (g) Rate (mg/min)
100 Chymosin (FPC) 6:17.23 2.29 1.16 1.13 3.136387039
200 Chymosin (FPC) 4.00.43 3.21 1.13 2.08 8.651418541

ANALYSIS

From the graph above it is clear that the 200L of 1 mg/mL of Chymosin FPC
produces more cheese per unit time, affirming the hypothesis. Some errors in conducting
this experiment might be using body temperature for incubation. Since human body has
an average, but has variances among different people, both solutions did not receive the
same amount of temperature, which could affect the curdling rate. This lab could be
improved by having more clearly recorded our results, as illegibility of the data could
skew the findings, making sure that weighing was under a standardized method. Some
further investigations that arise have to do with the amount of curds produced between
water and buttermilk, and whether the amount of milk affects the rate of curdling.

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 CHEESE LAB REPORT
Part III: Macromolecule Identification

Nihal Nazeem

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PURPOSE

To investigate what kinds of macromolecules are in cheese.

HYPOTHESIS

In string cheese monosaccharides, proteins, and lips will be present, as polysaccharides,

will not be present.

PROCEDURE

1. Benedict Solution (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 C)
d. Record all color changes
2. Lugols Iodine (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. Biuret Reagent (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. Lipids:
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.

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 b. Sudan IV Test
i. Add 120 microliters of Sudan IV solution to a 4 mL cheese sample.
ii. Gently mix.

DATA

Macromolecule Indicator Description of Positive Description of Negative Control

Control

Glucose Benedict's Solution Blue at the start Light blue

Yellow by 20s
Green by 33s
Completely Orange
by 60s
Red develops from
96 -120 seconds

Starch Lugols Iodine Black -ish Red- Dark Orange

Protein Biuret Reagent Purple Light blue

Fat I. Sudan IV I. Orange I. Red

II. Paper Towel II. Translucent II. Opaque
Test

ANALYSIS
String Cheese Macromolecules Tested
Macromolecule Presence Result Description

Glucose YES Change to green at top at

120 seconds with yellow
below, and some blue.

Starch NO Dark Orange; Brownish

Protein YES Violet; light-purple

Fat NO Pink; Semi- translucent

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 From the above table, String Cheese has glucose, and protein, without starch and
fat, disproving my hypothesis as I had included fat to be an item that is present in string
cheese. This could be as string cheese could have been made with/to have, a lower fat
content. Some errors in this lab could stem from the lack of the usage of Chymosin (FPC)
cheese. Since the sample was trashed by another class, we did the lab on a sample of
string cheese. Another issue was using 97 C rather than 100 C, for the control, and using
inconsistent temperatures, as the operation of the hot plate was lacking, for the actual
test, which could also have skewed results. The lab could have been improved by
including a separate procedure for making a cheese-water solution so that we can test
for cheese in a more standardized way, and using tongs to remove and place the tubes, so
that liquid isnt spilled as a reaction of heat and the human finger. In addition, testing
more samples to confirm our result could improve the lab, by providing accuracy. Some
further investigation this leads to deals with trying to understand what other substances
and organisms are in cheese, and how they affect what substances are present in cheese.

CONCLUSION

Cheese contains some macromolecules such as monosaccharides (glucose), and

protein (Casein), but not Polysaccharides (starch) and Lipids (fats), with Chymosin (FPC)
curdling agent-cheese being the fastest rate of amount of curds per unit time in
(mg/min). This was concluded by testing 100L of water, Buttermilk, Renin, and
Chymosin with with respective vials of 3 mL milk, and testing for macromolecules by
using a string cheese solution against the Benedicts Solution (monosaccharides), Lugols
Iodine (polysaccharides), Biuret Reagent (protein), the Paper Towel Test, and Sudan IV
(lipids). Chymosin came out to be the curdling agent with the highest curdling rate,
compared to the rest of the curdling agents when aggregated into the class average. In
the class average, Water had about 0.77 mg of curd per minute, as Buttermilk had 0.59
mg/min, Renin (NCB) had 7.4 mg/min, as Chymosin had 178.3mg/min. Since the highest
value is 178.3 mg/min, as the values of 0.77 mg/min, 0.59 mg/min, and 7.4 mg/min are all
comparatively lower, the highest number is linked to Chymosin (FPC). Thus Chymosin
has the highest rate of curdling per minute. When determining the macromolecules
present in cheese, the previously mentioned tests were used. When testing for the
control for the tests the positive controls are as follows: The Benedicts Solution had color
transformation from blue at the start to yellow by 20s, green by 33s, completely orange
by 60s, and the development of Red from 96 -120 seconds; Lugols Iodine was Black-ish;

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Biuret Reagent was purple; Sudan IV was orange, as the Paper Towel Test was
translucent. For negative controls: The Benedicts Solution had no color transformation
and remained to be blue; Lugols Iodine was red to dark-orange; Biuret Reagent was light
blue; Sudan IV was red, as the Paper Towel Test was opaque. When testing with a string
cheese solution, that was dissolved in boiling water, it had changed to green at top at 120
seconds with yellow below, and some blue for the Benedicts Solution; Lugol's Iodine was
dark orange to brownish; the Biuret Reagent was a violet; Sudan IV was pink, as the
paper Towel Test was semi-translucent. Since there was change in color for the
Benedicts Solution, that tested for glucose, the result was positive, meaning that the
string cheese contained monosaccharides. A dark orange color for Lugols Iodine, was
the negative control color, which matched the results, meaning there was no starch or
polysaccharides in string cheese. Since Violet was closer to purple positive control for the
Biuret Reagent that tested for protein, string cheese has protein. Finally, since Pink is
closer to the negative red for Sudan IV, and was semi-translucent for the paper towel
test, it had a very low amount of fat, but generally had no fat.

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