J.D.

TYTLER SCHOOL
CERTIFICATE
I Have The Pleasure To Certify That Khushboo
Nangia Student Of 12th–C, J.D.TytlerSchool
Has Persuaded His Work And Prepared The
Desertion On The Topic
“To Study The Factors Affecting Rate Of
Fermentation”
Under My Supervision And Guidance, This
Is Being Submitted For The Complete
Fulfilment Of The Requirement For Senior
Secondary Certificate.
Supervisor: Principal:
Dr.AzharAslam Khan.
(DepartmentOfChemistry)
Mrs.NeenaAndrews
(J.D.TytlerSchoolR‐
BlockNewRajinderNagarNewDelhi
ACKNOWLEDGEMENT
I take this opportunity of expressing my
deep gratitude and sincere regards to Dr.
Azhar Aslam Khan (Faculty of chemistry)
for his keen and personal interest,
guidance and moral support throughout
the entire accomplishment of work.
I thank our principal Ms. Neena Andrew
to provide us the permission and
appropriate equipment to complete this
project.
My sincere thanks to our lab assistant and
my friends for their extended cooperation
and assistance.

Yours sincerely,
Khushboo Nangia
 INDEX
• ACTIVITY OBJECTIVE
• THEORY
• REQUIREMENTS
• PROCEDURE
a) Substrate concentration
experiment
b) Temperature experiment
c) pH experiment
d) Substrate quality experiment
e) Yeast concentration experiment
• OBSERVATION TABLE
• RESULT
• PRECAUTIONS
• BIBLIOGRAPHY
ACTIVITY OBJECTIVE
To study the various factors affecting rate of
fermentation in yeast. These factors include pH,
temperature, nutrient availability, and the
concentration of available nutrients. By
determining which factors affect the yeast
activity, these variables can be controlled in the
fermentation process. This experiment will
illustrate that the growth of yeast is affected by
pH, temperature, and nutrient level and that one
natural by-product of this fermentation process is
carbon dioxide.
THEORY- FERMENTATION
Fermentation is a metabolic process that produces
chemical changes in organic substrates through
the action of enzymes. In biochemistry, it is
narrowly defined as the extraction of energy from
carbohydrates in the absence of respiration. In the
context of food production, it may more broadly
refer to any process in which the activity of
microorganisms brings about a desirable change
to a foodstuff or beverage. The science of
fermentation is known as zymology.
The term precision fermentation refers to
fermentation using precision biology (the latter of
which referring to the coming together of modern
information technologies (i.e. machine learning,
artificial intelligence, cloud‐based computing), and
modern biotechnologies as genetic engineering,
synthetic biology, metabolic engineering, systems
biology, bioinformatics, and computational
biology). It is a process that allows us to program
micro‐organisms to produce almost any complex
organic molecule. Precision fermentation is
particularly efficient for producing protein as well.
By producing it this way (in tightly‐controlled
environments, at optimized temperature), a
feedstock efficiency of 40%‐80% can be attained.
 Below are some definitions of fermentation. They
range from informal, general usages to more scientific
definitions.
1. Preservation methods for food via microorganisms .
2. Any process that produces alcoholic beverages or
acidic dairy products.
3. Any large‐scale microbial process occurring with or
without air.
4. Any energy‐releasing metabolic process that takes
place only under anaerobic conditions.
5. Any metabolic process that releases energy from a
sugar or other organic molecule, does not require
oxygen or an electron transport system, and uses an
organic molecule as the final electron acceptor .


The fermentation process of LAB starters can be
influenced by several factors such as temperature, pH,
strain capability, growth medium, inhibitors,
bacteriophage, incubation period, heat treatment of
milk, etc.
REQUIREMENTS
1. 125‐mL Erlenmeyer flasks or small
2. (8‐oz) glass soft‐ drink bottles
3. Balloons,7.8‐cm(7 inch)size
4. Table sugar(sucrose)
5. Brown sugar, jaggery , and glucose
6. pH paper
7. Wax pencil or marker
8. Large bottle or packages of rapid‐
rise yeast
9. Vinegar
10. Ammonia
11. Clock or Stopwatch
12. Warm water bath
13. Triple‐beam balances or scales
14. 100‐mL graduated cylinders
15. Thermometers
16. Incubator
 PROCEDURE
1-Substrate concentration experiment
Label flasks A through D. Add 80 mL of tap
water(neutral pH only)to each flask and
place the flasks in the following conditions:
Flask A - 0g sucrose
Flask B - 5g sucrose
Flask C - 10g sucrose
Flask D - 20g sucrose
Add 5g of rapid-rise yeast to each flask and
stir. Then place a balloon on each flask and
seal it securely with masking tape.
Periodically stir the contents by spinning the
flask slowly.
2-Temperature experiment
Label flasks A through D. Add 80 mL of tap
water(neutral pH only)to each flask and place
the flasks in the following conditions:
Flask E ~ in ice bath (0˚C)
Flask F- at room temperature (22˚C)
Flask G -in 40˚C water bath
Flask H- in 80˚C water bath
Dissolve 5g of sucrose in each flask. Add 5g of
rapid-rise yeast to each flask and stir. Then
place a balloon on each flask and seal it
securely with masking tape. Periodically stir
the contents by spinning the flask slowly.
3-pH experiment
Label flasks I through M. Add 80 mL of tap
water (neutral pH only) to each flask and add
vinegar or ammonia to adjust the pH as shown
below. Use pH paper to verify the pH.
Flask I- pH 2.
Flask J-pH 5.
Flask K- pH 7
Flask L- pH 9
Flask M -pH 11
Dissolve 5g of sucrose in each flask and warm the
solutions to 37˚C and add 5g rapid-rise yeast to
each solutions and stir. Then place a balloon on
each flask and seal it securely with masking
tape. Periodically stir the contents by spinning
the flask slowly.
4- Substrate quality experiment
Label flask N through Q . Add 80 mL of tap
water (neutral pH only) at 37˚C to each
flasks and dissolve 5g of each of the following
sugars:
Flask M - Sucrose
Flask N -Jaggery
Flask O - Brown sugar
Flask P-Glucose
Add 5g of rapid-rise yeast to each solution
and stir .Then place a balloon on each flask
and seal it securely with masking tape
Periodically stir the contents by spinning
the flask slowly.
5- Yeast concentration
experiment
Label flask R through V . Add 80 mL of tap
water (neutral pH only) at 37˚C to each flasks
and add 5g of sucrose in each of the following
flasks and add below mentioned amount of yeast :
Flask R~ 0g yeast
Flask S ~ 2g yeast
Flask T ~ 4g yeast
Flask U ~ 6g yeast
Flask V ~ 8g yeast
Then place a balloon on each flask and seal it
securely with masking tape Periodically stir the
contents by spinning the flask slowly.
OBSERVATION TABLE
SUBSTRATE EXPERIMENT
FLASK CONDITIONS FERMENTATION GAS
OBSERVED PRODUCED

A 37˚C +NO NONE LITTLE OR

SUROSE NONE

B 37˚C +5G SOME INITIALLY SOME

SUCROSE
C 37˚C +10G LITTLE LITTLE
SUCROSE
D 37˚C+20G THE MOST MOST
SUCROSE

(Observations after 20 minutes)

TEMPERATURE EXPERIMENT
FLASK CONDITIONS FERMENTATION GAS
OBSERVED PRODUCED

E SUCROSE + NONE LITTLE OR

ICE BATH NONE
F SUCROSE + LITTLE LITTLE
ROOM TEMP
G SUCROSE+ MOST SOME
40˚C
H SUCROSE + LITTLE THE MOST
80˚C

(Observations after 20 minutes)

 PH EXPERIMENT
FLASK CONDITIONS FERMENTATION GAS
OBSERVED PRODUCED

I 37˚C + PH 2 NONE LITTLE OR

NONE
J 37˚C + PH 5 SOME MOST

K 37˚C + PH 7 MOST SOME

L 37˚C + PH 9 LITTLE LITTLE OR

NONE
M 37˚C + PH 11 LITTLE LITTLE OR
NONE

(Observations after 20 minutes)

 NUTRIENT QUALITY
EXPERIMENT
FLASK CONDITIONS FERMENTATION GAS
OBSERVED PRODUCED

N 37˚C + SOME SOME

SUCROSE

O 37˚C + LITTLE LITTLE

JAGGERY

P 37˚C + BROWN LITTLE LITTLE

SUGAR

Q 37˚C + MOST MOST

GLUCOSE

(Observations after 20 minutes)

 YEAST CONCENTRATION
EXPERIMENT
FLASK CONDITIONS FERMENTATION GAS
OBSERVED PRODUCED

P 37˚C + 0G NONE LITTLE OR

YEAST NONE
R 37˚C + 2G LITTLE LITTLE
YEAST
S 37˚C + 4G SOME SOME
YEAST
T 37˚C + 6G SOME SOME
YEAST
U 37˚C + 8G THE MOST THE MOST
YEAST

(Observations after 20 minutes)

 IN THE SUBSTRATE QUANTITY EXPERIMENT,
FLASK D SHOWS THE MOST GROWTH AS
RATE OF FERMENTATION IS DIRECTLY
PRPORTIONAL TO THE SUBSTRATE BUT AS
THE AMOUNT INCREASES DUE TO SHORTAGE
OF YEAST RATE BECOMES CONSTANT.
 IN THE TEMPERATURE EXPERIMENT, FLASK
G SHOWS MOST GROWTH AS 40C IS THE
OPTIMUM TEMPERATURE FOR YEAST
GROWTH.
 IN THE PH EXPERIMENT FLASK K SHOWS
MOST GROWTH AS PH RANGE OF 6.5-7 IS
OPTIMUM FOR FERMENTATION IN YEAST.
 IN THE NUTRIENT QUALITY FLASK N AND Q
SHOWS BEST GROWTH AS BOTH ARE
COMPLEX SUGARS BUT GLUCOSE BEING
MONOSACCHARIDE SHOWS MORE
FERMENTATION IN LESS TIME.
 IN THE YEAST CONCENTRATION
EXPERIMENT FLASK R SHOWS MOST
GROWTH AS RATE OF FERMENTATION
INCREASES WITH INCREASE IN YEAST BUT
AS THE AMOUNT INCREASES DUE TO
LIMITING AMOUNT OF NUTRIENTS GROWTH
BECOMES CONSTANT.
CONCLUSION/RESULT

The Optimum Condition For

Yeast Fermentation Are As
Follows:
1) Absence Of Oxygen.
2) Temperature Range Of 20˚C
to 40 ˚C.
3) pH Range Of 6 to 7.5 .
4) Suitable amount of substrate
5) Sugar Contents Of Less Than
40% .
PRECAUTIONS
▪ Be careful when attaching balloons to
the flasks. Must carefully tape the
balloon securely to the flask to avoid any
leaks. Also, must remove the balloons
very carefully, as the foam, which will
rise into the balloons, will be expelled
under great pressure on removal. It is
suggested that the balloons remain
attached to the flasks until the next class
period to allow the carbon dioxide gas to
escape overnight.
▪ It is important that the tap water be
warmed to about 40˚C for each test prior
to adding the sugar and yeast. Keep
enough water available at this
temperature for all tests
BIBLIOGRAPHY
1.https://en.wikipedia.org/
wiki/fermentation

2.http://www.math.unl.edu/
~jump/Center1/Labs/What
%20Affects%20Yeast%20G
rowth.pdf

3.Google images