Alcohol Fermentation

What is the function of yeast?

Ans: Yeast plays a role as a leavening agent that is used in doughs or batters that cause a foaming action which
lightens and softens the mixture. Yeast breaks down sugars and converts it into carbon dioxide and alcohol that gives
volume to the bread.
Chemical equation
Ans: C6H12O6 (aq) ————> 2C2H5OH (aq) + 2CO2(g) + 210kJ
Word equation
Ans: Glucose (aq) ————> Ethanol (aq) + Carbon dioxide (g) + Energy
Possible experiment
1.

Procedure: -
1. Prepare two conical flasks and label them as P and Q. Add the following solution into each conical flask:
P: Room temperature 100ml distilled water + 30ml 5% glucose solution + 10g yeast
Q: 40°C (Manipulated variable) 100ml distilled water + 30ml 5% glucose solution + 10g yeast
2. Place a layer of paraffin oil on the surface of the solution and sealed a balloon at the mouth of both conical flasks
with sellotape. Make sure they’re sealed properly to prevent bursting of balloon. (Precautionary steps)
3. Measure (Responding variable) the diameter of the balloon with a thread first and mark it. Then, measure the thread
on a ruler. Record and tabulate the date.
4. Throughout the experiment, fix (constant variable) the volume of the distilled water, volume and concentration of
glucose solution and mass of yeast. To prevent parallax error, measure the length of the thread, volume of distilled
water and glucose solution by placing the eye vertically above the mark being read or read the bottom of the meniscus
for concave liquid surface.
Observation: - (State value & unit, choose the greatest or lowest one)
The diameter of the balloon at 20th minute for conical flask Q is 16.5cm.
Inference: -
The fermentation of yeast is higher at optimum temperature.

Variables Ways to handle variable

Manipulated variable: - Use different temperature for each solution in conical
Temperature of the solution flask P and Q

Responding variable: - Measure the diameter of the balloon using a thread first
Diameter of the balloon and mark it. Then, measure the thread on a ruler.

Fixed variable: - Fixed the volume of the glucose solution as 30ml and its
Volume and concentration of glucose solution concentration as 5%.

Hypothesis: -
The balloon sealed at conical flask Q has higher diameter compared to the balloon sealed at conical flask P.
Graph: - (The graph title must be written as “<Y-axis label> versus <X- axis label>”. )

Prediction question: - (Must state value and unit and explanation)

Repeated with 20g yeast
Diameter of balloon is bigger than 16.5cm that is 18.2. This is because the increase of mass of yeast will increase the
enzyme zymase/ rate of yeast fermentation.
2.

Procedure: -
1. Prepare two boiling tube and label them as A and B. Add the following solution into each boiling tube:
A: 20ml 5% glucose solution that is boiled and cooled down + 5ml yeast suspension (Manipulated variable)
B: 20ml 5% glucose solution
2. For both boiling tube, place a layer of paraffin oil on the surface of the solution and covered with a perforated cork
stopper with a delivery tube inserted where its end is inserted to a test tube filled with 2ml of limewater. Make sure the
end of the delivery tube is submerged into the limewater. (Precautionary steps)
3. Record the initial temperature and left the set up for X time. After that, record the changes (Responding variable) to
the temperature and the change in the appearance of limewater as well as the smell emitted from the test tubes.
Tabulate the data.
4. Throughout the experiment, fix (constant variable) glucose solution make sure it is cooled down. To prevent
parallax error, measure the temperature by having the eye perpendicular to the point of the scale
3.

Procedure: -
1. Prepare two boiling tube and label them as A and B. Add the following solution into each boiling tube:
A: 15ml 5% glucose solution that is boiled and cooled down (Manipulated variable) + 2g yeast
B: 15ml 10% glucose solution that is boiled and cooled down + 2g yeast
2. For both boiling tube, place a layer of paraffin oil on the surface of the solution and covered with a perforated cork
stopper with a delivery tube inserted where its end is inserted to a test tube filled with 2ml of limewater. Make sure the
end of the delivery tube is submerged into the limewater. (Precautionary steps)
3. Start the stop watch and record (Responding variable) the number of air bubbles released in x time by using
stopwatch. Tabulate the data.
4. In this experiment, stir the mixture using a rod slowly until mix and avoid over stirring and fix the mass of the yeast
(constant variable).
Correction
More air bubble release. In the presence of oxygen yeast carries out aerobic respiration.
C6H12O6 (aq) ————> 2C2H5OH (aq) + 2CO2(g) + 210kJ
C6H12O6 (aq) + 6O2 (g) ————> 6CO2(g) + 6H2O+ 210kJ
Problem Statement:
What is the effect of volume of glucose on change of diameter and height of dough?

Aim:
To study the effect of volume of glucose to change of diameter and height of dough.

Inference:
More glucose used in fermentation to produce more carbon dioxide to rise the diameter and height of bread dough.

Variables:
Manipulated variable, MV : Volume of glucose
Responding variable, RV : Change of diameter and height of dough
Constant variable, CV : Mass of wheat flour and yeast

Hypothesis:
The higher the volume of glucose, the higher the rate of fermentation.

Procedure:
1. Measure 100g of wheat flour in a beaker.
2. Put 1g of yeast in beaker and add 10ml distilled water. Stir mixture with glass rod, leave mixture for 10
minutes.
3. Add mixture of yeast into 100g wheat flour and add sufficient water to make bread dough in basin.
4. Measure initial diameter and height of bread dough, coat dough with oil. Record data.
5. Cover bread dough with cloth.
6. After 15 minutes, measure the final diameter and height of bread dough. Record data.
7. Calculate change of diameter and height of dough.
8. Calculate the rate of change of diameter and height of bread dough. Record data.
9. Repeat experiment by using 10ml glucose solution and 20ml glucose solution in making the bread dough.

Tabulation of data:
By diameter and height
Volume of Diameter of bread dough Height of bread dough Rate of Rate of
glucose change of change of
solution Initial (cm) Final (cm) Initial (cm) Final (cm) diameter height
(ml) (𝑐𝑐𝑐𝑐 𝑚𝑚𝑚𝑚𝑚𝑚−1 ) (𝑐𝑐𝑐𝑐 𝑚𝑚𝑚𝑚𝑚𝑚−1 )
0 7 10 2.5 3 0.20 0.03
10 8 11 2.5 3.5 0.20 0.07
20 7 12.5 3 5 0.37 0.13

Bread dough solution in measuring cylinder

Volume of glucose
Volume of bread dough solution (c𝑚𝑚3 ) Rate of change of
solution (ml)
volume (c𝑚𝑚3 𝑚𝑚𝑚𝑚𝑚𝑚−1 )
Initial Final

Observation:
For bread dough with 20ml glucose, the initial diameter is 7, the final diameter is 12.5 and the initial height is 3, the
final height is 5.

Operational definition:
Define, State quantity of MV CV RV (choose any experiment)
(Fermentation the incomplete breakdown of glucose in limited or no oxygen condition to release carbon dioxide,
ethanol and 210KJ energy/ Cellular respiration is the oxidation of glucose through several stages to release energy/
Aerobic respiration is the complete breakdown of glucose in presence of oxygen to release carbon dioxide, water and
2892KJ energy/ Glucose is the main substrate for cellular respiration/ Glycolysis is the breakdown of glucose ) when
20ml of glucose solution is added into dough with 1g of yeast, water and100g wheat flour to increase the diameter of
dough from 7 to 12.5 and height from 3 to 5.

Extra questions:
1. Experiment is repeated by exposing bread dough with 10ml glucose in the air after kneading. Predict and
explain the rate of change of height of bread dough after 15 minutes.
Rate of change of height of bread dough after 15 minutes lower than 0.07 that is 0.03. Exposed to air cause
evaporation of water vapour in dough, dough outer surface harden constrain the expansion of dough./ Water is
necessary for yeast fermentation and reproduction in producing more carbon dioxide to raise the bread.
Problem Statement:
Which bread dough have highest change of diameter and height?

Aim:
To study the effect of presence of yeast to change of diameter and height of dough.

Inference:
Presence of yeast carry out fermentation, producing carbon dioxide to raise the bread dough, changing the diameter
and height of dough.

Variables:
Manipulated variable, MV : Presence of yeast
Responding variable, RV : Change of diameter and height of dough
Constant variable, CV : Mass of wheat flour

Hypothesis:
Dough with presence of yeast has highest change of diameter and height compared to dough without yeast.

Procedure:
1. Measure 100g of wheat flour in a beaker.
2. Put 1g of yeast in beaker and add 10ml distilled water. Stir mixture with glass rod, leave mixture for 10
minutes.
3. Add mixture of yeast into 100g wheat flour and add sufficient water to make bread dough in basin.
4. Measure initial diameter and height of bread dough, coat dough with oil. Record data.
5. Cover bread dough with cloth.
6. After 15 minutes, measure the final diameter and height of bread dough. Record data.
7. Calculate change of diameter and height of dough.
8. Calculate the rate of change of diameter and height of bread dough. Record data.
9. Repeat experiment by using distilled water only in making the bread dough.

Tabulation of data:
Temperature Height of bread dough Rate of Rate of
Presence of change of change of
yeast Initial (cm) Final (cm) Initial (cm) Final (cm) diameter height
(𝑐𝑐𝑐𝑐 𝑚𝑚𝑚𝑚𝑚𝑚−1 ) (𝑐𝑐𝑐𝑐 𝑚𝑚𝑚𝑚𝑚𝑚−1 )
Present 7 10 2.5 3 0.30 0.05
Absent 7 7 4.5 4.5 0 0

Observation:
Bread dough with yeast have increase in diameter from 7 to 10, height from 2.5 to 3 while bread dough without yeast
have remained the same diameter 7 and height 4.5.

Operational definition:
(Fermentation the incomplete breakdown of glucose in limited or no oxygen condition to release carbon dioxide,
ethanol and 210KJ energy/ Cellular respiration is the oxidation of glucose through several stages to release energy/
Aerobic respiration is the complete breakdown of glucose in presence of oxygen to release carbon dioxide, water and
2892KJ energy/ Glucose is the main substrate for cellular respiration/ Glycolysis is the breakdown of glucose ) when
1g yeast is added into 100g wheat flour and water to increase the diameter of bread dough from 7 to 10 and height
from 2.5 to 3.

Extra questions:
1. Experiment is repeated by adding 10ml boiling distilled water to the yeast. Predict and explain the rate of
height change for the bread dough after 15 minutes.
Rate of height change for bread dough after 15 minutes lower than 0.05 that is 0. Boiled water, extreme
temperature denatures the zymase, zymase unable carry out cellular respiration release carbon dioxide to raise
the bread dough.
Procedure:
1. Put 1 g of wheat flour in a test tube and add 5mL of distilled water.
2. Conduct iodine test and Benedict's test for wheat flour solution.
3. Repeat the experiment by using 5mL 5% glucose solution and potato.

Variables:
Manipulated variable, MV : Type of food sample
Responding variable, RV : Colour change in iodine and benedict test
Constant variable, CV : Volume of iodine and benedict solution

Inference:
Potato and wheat solution contain starch, glucose solution absent in starch. Wheat and glucose contain reducing sugar,
potato absent in reducing sugar.

Tabulation of data:
Type of food sample
Wheat flour solution
5ml 5% glucose solution
Potato
Colour change for iodine test Colour change for Benedict’s
test
Yellow to dark blue Blue to orange
Yellow remains unchanged Blue to brick red
Yellow to dark blue Blue to orange

Benedict test:
1. 2ml benedict solution + 2ml sample (ratio 1:1)
2. Boil water
3. Water bath 100 °C for 5 min

Iodine
1. 2 drop iodine + 2 drop sample (ratio 1:1)