GROUP No.
___ SECTION
Date:____
___________________________________________________________________
TITLE:
FERMENTATION
INTRODUCTION: Fermentation is a core biological process in which microorganisms, such as yeast and
bacteria, convert sugars into simpler compounds like alcohol, gases, or acids under anaerobic conditions.
In modern biology, fermentation is studied not only for its historical role in food preservation and alcohol
production but also for its relevance in biofuel production, metabolic engineering, and industrial
biotechnology. This experiment introduces engineering students to metabolic pathways, enzyme
action, and bioprocess parameters using a simple fruit fermentation model
PROBLEM/OBJECTIVE:
> Determine how sugar concentration and temperature affect the rate and extent of fruit fermentation
> To be able to produce fermented product with varying indepdendent variables
THEORY: Fermentation occurs when microorganisms like yeast convert sugars (glucose, fructose,
sucrose) into ethanol and carbon dioxide. The process is sensitive to environmental conditions such as:
Sugar Concentration and Temperature.
In fruit fermentation, the natural sugars and nutrients serve as substrates for yeast, leading to the
production of ethanol and CO₂. By adjusting the concentration of sugar and incubation temperature, we
can simulate a controlled bioprocess and monitor fermentation activity.
MATERIALS AND SET-UP
Materials:
Sugar and Yeast Fermentation Experiment Anaerobic Fermentation with
Balloons
You may refer to this video link
Ripe fruit juice (e.g., apple, mango, or pineapple) – 500 mL
Active dry yeast – 1 packet (~7g)
Table sugar (sucrose)
Warm water and Ambient Temperature water (you could use cold)
4-liter plastic bottles or glass jars with balloon tops or fermentation airlocks
Thermometer
Measuring cups and spoon
Labels and marker
pH strips (optional)
Digital kitchen scale
Timer or clock
METHODOLOGY: (step by step procedure done by students with image)
1. Prepare Fruit Juice:
Use 500 mL of Blended filtered or strained juice in each container.
2. Add Sugar:
>To each jar, add sugar according to your variable: (if you don’t have weighing scale you may use
tablespoon and ratio with the jar, take note of how many you used and observe proper measurement)
0 g (control)
25 g (5% w/v)
50 g (10% w/v)
And another 25 g 5% for ambient temperature water (could use cold water)
Stir until dissolved.
3. Activate Yeast:
In a separate cup, dissolve 7g of yeast in 50 mL warm water (~35°C) and let sit for 10 minutes until
foamy. Prepare this three times with warm water then prepare a fourth with ambient temperature water
(could use cold water)
4. Combine:
Pour yeast mixture into each juice + sugar mix and stir.
5. Seal Container:
Stretch a balloon over the mouth of the bottle to trap gases (or use an airlock if available).
6. Incubate:
Place half the samples at room temperature (~25°C), and half in a warm place (~35°C).
Label containers with sugar level and temperature condition.
7. Monitor:
Observe balloon inflation daily.
8. Record smell, balloon size, and any visible changes (e.g., bubbling).
Every day for 4 days after 24 hrs
DATA AND RESULTS: (with computations/ simulations)
QUESTIONS and ANALYSIS OF DATA:
1. What metabolic pathway is responsible for fermentation in yeast?
2. How does increasing sugar concentration affect fermentation rate and product yield?
3. What is the optimum temperature for yeast fermentation and why?
4. How could pH changes during fermentation affect microbial activity?
5. What challenges would scale-up introduce in a real bioreactor setting?
6. What safety precautions must be considered in large-scale fermentation?
7. How would you design a low-cost sensor to monitor CO₂ production in real time?
8. How do fruit type and composition influence the fermentation process?
9. Can the resulting ethanol content be estimated based on gas production? How?
10. What are the implications of microbial fermentation in renewable energy and sustainable
engineering?
CONCLUSION: Group answer includes the answers to the problems
(Include a brief statement as a group that will conclude the experiments, and small
concluding statements for the student)
Experiment No. 3 Fermentation
Fermentation is a metabolic process that converts sugars to acids, gases, or alcohol in the
absence of oxygen. It is a crucial biochemical reaction carried out by microorganisms such as
yeast and bacteria. One of the most well-known fermentation processes is alcoholic fermentation,
where yeast (Saccharomyces cerevisiae) converts glucose into ethanol and carbon dioxide. This
process is widely utilized in the production of alcoholic beverages, bread, and biofuels.
The study of fermentation dates back thousands of years, with ancient civilizations using it to
produce beer, wine, and leavened bread. In modern times, fermentation has become a
cornerstone of the biotechnology industry, contributing to the production of a variety of
products, including antibiotics, enzymes, and vitamins.
During fermentation, yeast cells metabolize sugar in the absence of oxygen, leading to the
production of ethanol and carbon dioxide. The general reaction for alcoholic fermentation is:
C6H12O6→2C2H5OH+2CO2
In this experiment, students will explore the fundamentals of fermentation by observing the
activity of yeast in a sugar solution. By measuring the production of carbon dioxide, an
observable by-product of fermentation, students will gain insight into the dynamics of this
metabolic process. They will also investigate how various factors, such as temperature and sugar
concentration, affect the rate of fermentation.
Materials
Fruits for fermentation – ginger , apple , pine apple, banana ANY, good for 3 samples
(Peeled na preferably, calamansi , lime lemon )
Wine yeast: 1/2 packet (2-3 grams).
Distilled water enough to fill 3 glass bottles
Sugar: Optional, to increase alcohol content (up to 100 grams).
Airlock: Straw, plastic and rubber band to seal and have a hole for CO2 to escape
Fermentation vessel: A clean pasteurize bottle , 250 , 500 or 1 liter any. 3 pieces preferably
same volumes
Aluminum foil – to ensure the sample is in a dark environment/ and away from sunlight.
OBJECTIVE:
The students under the experiment will
Understand the Fermentation Process:
Develop Practical Laboratory Skills:
Analyze and Interpret Experimental Data:
Understand Applications of Fermentation:
METHODOLOGY
Sanitize Equipment:
May be done at home, Pour boiling water on the glass, pasteurize the container for 3-5
minutes.
Prepare Must:
Slice the fruit, skin peeled, and put it on the container
Pour Distilled water to hydrate the fruits
If desired, dissolve up to 100 grams of sugar in the container to increase the potential
alcohol content.
Add Yeast:
Hydrate the wine yeast according to the package instructions, measure the amount of
yeast in ratio to the volume of the sample added then add it to the fruit juice
Now fill the container into the desired amount and get ready to seal
Attach Airlock:
Seal the container with a plastic lid, punch a hole for a straw, the straw can be CUT to a
smaller size,
Primary Fermentation:
Place the vessel in a dark, cool place (around 20°C or 68°F) and let it ferment for about 1
week. Bubbles in the airlock will indicate active fermentation.
Secondary Fermentation and Aging:
Once bubbling slows significantly, transfer the wine to a clean vessel (anybottle) to leave
behind sediment. Attach the airlock and let it age
Sampling
Observe the sample during initial preparation, 1 week of fermentation and the 1 week of
primary fermentation fermentation, get a sample of the wine every 3 days and observe its
quality, Color smell, description,
Do this for all three trials, Three trials may differ in fermentation factors - volume –
sugar – yeast – temperature - (kayo na bahala preferably hindi lahat ng grupo ay
magkakaparehas)
Questions , Data and Analysis (in these experiment sharing of data is encourage)
1. How did the temperature of the environment affect the rate of fermentation? If the
experiment is repeated at different temperatures, what differences are observed in the
rate of carbon dioxide production?
2. How does the amount of yeast used affect the rate of fermentation? If you were to
double the amount of yeast, what changes would you expect in the carbon dioxide
production?
3. How does the length of time the yeast is allowed to ferment affect the total amount of
carbon dioxide produced? What is the relationship between fermentation time and gas
production?
4. Describe any visible changes in the fermentation mixture over time. What signs indicate
active fermentation, and how do these signs change as the fermentation progresses?
5. How can the results of this experiment be applied to industrial fermentation processes?
What factors need to be controlled to optimize fermentation in commercial settings?
6. If the fermentation were allowed to continue for a longer period (e.g., several weeks),
what changes would you expect in the rate of carbon dioxide production and the
characteristics of the fermentation mixture?
