Situation: Sarah makes homemade cheese as a way of saving money.

She uses the enzyme rennin to

coagulate the milk and then strains the whey out, leaving the curds behind. She then uses the curds to
make her homemade cheese. Sarah’s friend, Jane, suggested that she add a bit of acid to the mixture of
milk and rennin as a means of getting even more curds from the milk. Jane suggests that this may
increase the activity of the enzyme rennin. Plan and design an experiment to test the reliability of Jane’s
suggestion.

Hypothesis: the enzyme rennin works best in an acidic environment.

Aim: to determine if the enzyme rennin works best in an acidic environment compared to a non-acidic
environment.

Apparatus/Materials: 1 litre of milk (1000ml), junket rennet tablet box (containing 10 tablets that are
made from the enzyme rennin), 10 500ml beakers, cheesecloth, beam scale, measuring cylinder, 1 large
bowl.

Method:

1) Obtain 1 litre of milk.

2) Strain all the milk to remove any curds that may have been there before by pouring it into the cheese
cloth and fold the top of the cloth to surround the milk inside. Squeeze the cheesecloth for 5 minutes
over a sink. Then pour the milk into a large bowl.

3) Label the 10 beakers A1, A2, B1, B2, C1, C2, D1, D2, E1 and E2 respectively.

4) Add 100ml of milk to each beaker by using the measuring cylinder

5) Prepare 6 buffered milk solutions with a pH of 3 in A1 and A2, 4 in B1 and B2, 7 in C1 and C2, 9 in D1
and D2, respectively for each pair.

6) Add 1 tablet of rennet tablet, which contains enzyme rennin, to each of the 10 beakers respectively.

7) Immediately set the solutions down in the same room and let it rest for 2 hours.

8) After the 2 hours is up, strain out the curds in beaker A1 by pouring the milk solution in the
cheesecloth and squeezing for 5 mins to get all the moisture out, leaving the curds wrapped in the cloth.
This is done over a sink because the milk would spill.

9) Weigh the curds collected in the cheesecloth on the beam scale and record it.

10) Repeat steps 8 and 9 for all the remaining beakers and record each weight respectively.

11) Make a table showing the beakers, pH level used for each beaker, mass of curd/g collected from
each beaker and the average mass/g from the pair of beakers.
Controlled variables: amount of milk in each beaker, the temperature of the milk solutions when left to
rest for 2 hours and the amount of rennin used.

Manipulated variables: pH of milk.

Responding variables: weight of curds collected from each beaker.

Results: Table showing productivity of enzyme rennin at different pH values.

Beaker pH Mass of curd/g Average Mass/g

A1 3
A2 3
B1 4
B2 5
C1 7
C2 7

D1 9

D2 9

E1 N/A

E2 N/A

Expected results: The enzyme rennin is found in the stomach in some animals and its optimum pH level
is 3.4. Therefore it means that a pH level close to 3.4 would show more activity of the enzyme rennin
and that of a higher pH level. After pH level 7 (neutral) it rapidly loses its productivity. The enzyme
rennin is a protein-digesting enzyme and it causes curdling in milk by transfroming caseinogen into
insoluble casein.
Assumption: Rennin will cause the milk to curdle. The amount of curds was a result of the activity of the
rennin.

Precaution:

1. Avoid error due to parallax when using the measuring cylinder by measuring at eye level.

2. Take care when pouring the milk into the beaker so no spillage takes place.

Source of error:

1. Some curds may have passed through the cheesecloth because it was too small and this could
have caused inaccurate results.

2. When squeezing the cheese cloth it may not have been the same pressure applied throughout
the entire of the 5 minutes.

3. When pouring the milk measured in the measuring cylinder to the beaker, droplets could have
remained in the measuring cylinder.

Limitation:

1. Some of the curdling may have been caused by the acidic pH because that also causes curdling
in milk.

2. Small amounts of curdles may have seeped through the cheese cloth resulting in inaccuracy in
results.