Cheese Lab

By: Holly Jossart

Purpose:

The reason we performed the cheese lab was to understand macromolecules through

cheese and which ones are in cheese, to learn how cheese is made, and what variables

help cheese produce the fastest.

Hypothesis:

Part 1 If we test the fermenting agents FPC, NCB, Buttermilk, and water, then FPC will

be the most efficient.

Part 2 If we test different fermenting agent amounts to the milk, then there will not be a

time difference in how long the milk takes to curdle.

Part 3 If we test our cheese for starch, protein, lipids, and glucose, then the cheese will

contain protein, lipids, and glucose.

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 100 l 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.
Part 2
1. Label the four 6ml with the amount 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 100, 200, 300, and 400 l of fermentation
produced chymosin (FPC).
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.
Part 3
- Monosaccharide/Glucose
1. Obtain a vial, and place a cheese sample into it, that is of the approximate
volume of 5mL.
2. Into this vial, pipet 5 mL of Benedict's solution. Mix well.
3. Heat for 2 minutes in a boiling hot water bath (100 mL of water in a 250-mL
beaker at 100 degrees celsius)
4. Record all color changes
-Polysaccharide/Starch
1. In a test tube, mix 5 mL of cheese sample with 0.625 mL of Lugols iodine.
2. Gently swirl to mix. Do not heat.
3. Record all color changes.
-Protein
1. Place 4 mL of a cheese sample in a test tube.
2. Add 1.5mL of Biuret reagent to the test tube.
3. Mix well.
4. Record the color change after 30 seconds.
-Paper Test
1. Melt the cheese in a test tube by inserting the vial in a heated water bath.
2. Pour the melted cheese onto a piece of paper.
3. After waiting for the cheese to dry and disperse, hold the paper to light.
4. Record the percentage of translucence.
-Sudan IV Test
 1. Add 120 microliters of Sudan IV solution to a 4 mL cheese sample.
2. Gently mix. Record color change.
Data

Part 1

Agent Curdling Weight of Weight of Weight of Rate

Time (min) Cone & Cone (g) Curds (g) (mg/min)
Curds (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

- The FPC agent curdled in 30 minutes, while the other three curdled in around 24

hours (overnight).

- Every sample had a white coloration and smelled of sour milk.

- FPC was the most solidified compared to the others agents.

Part 2

Amount of Curdling Weight of Weight of Weight of Rate

FPC (l) Time (min) Cone & Cone (g) Curds (g) (mg/min)
Curds (g)

100 3 1.39 1.15 0.24 80

200 3 1.37 1.15 0.22 73.33

300 3 1.45 1.15 0.3 100

400 3 1.37 1.15 0.22 80

- The amount change of the FPC agent did not affect the curdling time of the milk.
 - The weight of the curds and cone and curds varied slightly.

Part 3

Indicator Testing (before testing cheese)

Standard Indicator Description of Positive Description of

Control Negative Control

glucose Benedicts Solution 20s.- turning orange light blue

45s.- spreading
60S. -completely orange

starch Lugols Iodine dark brown light orange/brown

protein Biuret Reagent dark blue light blue

fat Sudan IV or paper reddish pink clear

bag test
- protein, fat, and glucose had positive results, while starch did not have any

apparent changes.
 Indicator Testing (before testing cheese)

Glucose Starch

Positive (left), Negative (right) Positive (left), Negative (right)

Protein Fat

Positive (left), Negative (right) Negative (left), Positive (right)

 Indicator Testing (with cheese)

Standard Indicator Cheese type Results Control

used

glucose Benedicts FPC blue to green Positive

Solution over 2 min.

starch Lugols Iodine FPC dark brown Negative

protein Biuret Reagent FPC dark blue Positive

fat Sudan IV or FPC clear Positive

paper bag test

Cheese Testing for Control

Order from left to right: Glucose, Protein, Starch, Fat

Analysis

Part 1

In the first part of the lab, making the cheese with 4 different agents, our table and

graph above prove that FPC is the most efficient curdling agent, timewise. From most to

least efficient, the agents are FPC, NCB, buttermilk, and water. The higher the rate is,

the faster the cheese curdled with each agent.

Our hypothesis was correct because FPC was the most efficient, according to our

testing and the resulting data.

Errors in the lab mainly were because of lab equipment and the way we took

measurements. The lack of racks and test tube stoppers caused problems with leaking

milk, changing the amounts of curdled milk we ended with. This caused an improper

measurement of the milk curds along with the fact that it was hard to remove all the
curdles from the test tubes. With losing bits of the samples in two different ways, our

measurements varied, meaning our data also varies where it shouldnt.

To improve the lab, more lab equipment should be used to ensure we dont lose parts of

the sample (stoppers and test tube racks) and a different method to removing the milk

curds should be considered.

Further testing encouraged by this lab could be to find how different quantities of milk or

the agents can affect the curdling time and if FPC is always the most efficient even with

different amounts.

Part 2

From the graph and table, 300 l is the most efficient amount of FPC to milk ratio. But,

because the other results vary much differently, this was probably due to human error.
Our hypothesis was incorrect because there were different time differences based on

our data. But, we do not know 100% if our hypothesis is wrong because there was

human error in our data for the 300 l.

A major error was with the 300 l test. Although we dont know the exact error, we know

there was one that gave us inaccurate data. The error may have been right from the

start and we may have added too much or too little FPC. It also could be because of

spilling or incorrect measurements. Because we incubated in our armpits, we may have

lost some of the sample while holding it.

The lab could be improved by using a real incubation chamber rather than our armpits.

Although our armpits are all supposed to be room temperature, it can vary if someone is

cold, sick, or moving more than others. And because we used four different armpits, the

chance of human error is much higher.

For future investigations, we could look into how different temperatures affect the

curdling. If we make it colder or warmer, will the curdling time increase, decrease, or

stay the same. We also could see if less amounts of FPC will affect how the milk

curdles, knowing that more does not change anything.

Part 3

Based off the second data table for part 3, our cheese tested positive for glucose,

protein, and fat, and negative for starch.

Our hypothesis was correct according to our testing and resulting data.

For this lab, there were no apparent errors as the data we found was accurate.
One thing that could be improved though, would be a different method for testing the

cheese. While we smashed the cheese as much as we could, it would have been easier

to mix everything if we had grinded the cheese into a fine powder. We did not have the

equipment to do so, but with a mortar and pestle, we could have.

Further testing inspired by this lab could be to see what other food have in them. For

example, seeing if bread has glucose, protein, fats, or starch.

Conclusion

Cheese contains glucose, proteins, and fats and can be produced most efficiently by

300 l of FPC for 3 ml of milk. In this experiment, we started off by testing which

curdling agent was the most effective in making cheese. We then tested how different

amounts of that agent affected the cheese curdling process, and finally, we finished by

testing what macromolecules are in cheese.

From our data, we know that 100 l of FPC is the most efficient way to curdle milk. This

is because it curdled the fastest, only taking 5.65 minutes while the other agents took

around 24 hours to curdle. The reason for this is because FPC is engineered to curdle

milk fast. It is the same thing as NCB, but NCB is natural, while FPC is altered.

Buttermilk and water are usually not used for curdling milk over FPC.

Based on the second part of the lab, we know that the amount of FPC used in relation

to the amount of milk is irrelevant. In our data, each different amount of FPC took 3

minutes to curdle. And although the 300 l of FPC was an outlier in the data (much

higher rate, 100 mg/min) we are excluding it because we believe there was an error in

the process. Otherwise, the rates for 100, 200, and 400 l were all around 80 mg/min.
Variation can occur because of inaccurate measuring. Although the amount of FPC

does not matter, 100 l is the most efficient because it uses the least amount of FPC.

Finally, we found that cheese contains protein, glucose, and fat. Our data proves it

using multiple tests that activate color change when a certain macromolecule is present.

The reason for there being these macromolecules is because when the milk curdles to

cheese, strains of the macromolecules form. For example, we know that the cheese has

glucose in it because it went through a test using Benedicts solution and the color

changed from blue to green, meaning it was glucose positive.