RENNET LAB EXTENSION

AP BIOLOGY 4B
Connor Chen, Daniel Xiao, Joon Guzman, Julien Dintino, Matheus Siquiera
Background: An enzyme’s ability to catalyze substrates is highly dependent on temperature. To test how
temperature affects enzyme activity in this experiment, we will be using rennet, an organic extract that
primarily contains the enzyme chymosin/rennin. Rennin coagulates protein (casein) in milk through a
catalysis reaction. Prior to designing this experiment, we recorded the qualitative effects of rennet on
milk. When added to dairy milk with a temperature within the optimal range of rennet, it cleaves the
casein enzymes, making them bind to each other. This process forms a gel of casein protein that separates
from the milk whey (liquid portion of milk). By changing the temperature of the rennet’s environment, we
will record how temperature affects enzyme activity through the measuring the mass of curds produced
during coagulation.

Qualitative Data Table:

Substance: Milk Only Milk +Acid Milk + Rennet Milk + Rennet + Acid

Observations: The consistency The resultant was The liquid was The resultant was curd, a
of the milk slightly viscous, moderately relatively solid substance
stayed the same, with a few viscous, much resembling cottage
as it remained clumps of solids thicker in cheese.
fully liquid. formed. consistency than
that of acid+milk.

Research Question: How does changing the temperature of milk affect the enzyme activity, measured
through the mass of milk curds produced.

Independent Variable: Temperature of milk (20C, 30C, 40C, 50C, 60C).

Dependent Variable: Mass of milk curds (g)

Controlled Variables:
Controlled Variables Controlled Variable 1: Controlled Variable 2: Controlled Variable 3:
Amount (volume) of Type of milk Time stirred
rennet solution added
Reasoning The volume/amount of The type of milk used A controlled mixing
the rennet is controlled in this experiment can time ensures that milk
because if the rennet cause varying results as and rennet samples are
enzyme varies among some milks such as under or over mixed.
different groups the rate whole milk are mostly The quantitative
of reaction could be consisted of saturated observation of the milk
 lower of greater for fats that can cause may vary if mixing
depending on the different kinetic speeds time is uncontrolled
saturation of rennet. of the enzymes and for.
Controlling rennet substrates. Chemical
solution ensures that differences in plant and
the number of enzymes dairy milks also
available for catalysis. respond differently to
the rennet enzyme. One
last key factor of the
differing milks is the
changes in pH
Method This variable is This variable is This variable is
controlled by using a controlled by using the controlled by using a
graduated cylinder. same type of milk from handheld blender and a
This lab appliance will the same bottle of milk timer. The handheld
enable experimenters to throughout the blender can mix the
measure the rennet experiment. This solution at a constant
solution in a accurate prevents other factors rate, and the timer
fashion. in the milk, such as the ensures that all
density of sugar, fat solutions are blended
content, or casein for a constant amount
presence from of time. This ensures
impacting the that different blending
experimental results. times will be
minimized and the
experimental results
will be kept constant.

Materials:
-1.5 liters of full fat milk
-Stirring stick
-Scale (in grams)
-Beaker
-Rennet solution
-Hot plate
-Strainer
-Bowl/petri dish
-Thermometer
-Timer/stopwatch
-Safety goggles
-Gloves

Methodology:
1. Find a climate-controlled room and fill a beaker with 100ml of room-temperature (20ºC) full fat
milk.
2. Add 5 drops of rennet solution to the beaker.
3. Stir the solution for 15 seconds and begin timing.
 4. 5 minutes after the rennet is added, pour the contents of the beaker through a strainer into a bowl
or petri dish, separating the curds from the whey.
5. After discarding the milk, place the curds from the strainer on a scale.
6. Measure the mass of the curds, and enter the data into your table.
7. Wash out the beaker, petri dish/bowl, and strainer thoroughly.
8. Repeat steps 1-7 but heat the milk to 30, 40, 50, and 60 degrees Celsius by placing the beaker on
a hot plate before adding the rennet.
9. Repeat steps 1-8 two more times so you have a total of 15 data points (3 for each temperature).3

Hypothesis: As temperature increases, the mass of the milk curds produced by adding rennet to milk will
increase. However, after around 40ºC, increasing the temperature will decrease mass of the milk curds
produced.

Predicted Data:
Effect of Temperature on Mass of Milk Curds

Mass of milk Temperature (oC)

curds (g) 20 (control) 30 40 50 60
Trial 1 5.1 9.6 20.0 4.5 2.1
Trial 2 4.4 10.0 19.6 3.6 1.8
Trial 3 4.4 9.6 22.1 5.0 2.2
Average 4.6 9.7 20.6 4.4 2.0

Predicted Graph: