Bi 6 Biotechnology for Everyone, Laboratory

Fermentation

Yogurt is a fermented milk product produced by adding a "starter" of active yogurt containing
a mixed culture of Lactobacillus bulgaricus and Streptococcus thermophilus. These bacteria produce
lactic acid during fermentation of lactose. The lactic acid lowers the pH, makes the milk tart, causes
the milk protein to thicken and acts as a preservative since pathogenic (disease-causing) bacteria
cannot grow in acid conditions (with lids intact, this yogurt will keep at least a month or two in the
refrigerator). The partial digestion of the milk when these bacteria ferment milk makes yogurt easily
digestible. In addition, these bacteria will help settle gastrointestinal tract upset including that which
follows oral antibiotic therapy by replenishing non-pathogenic flora of the gastrointestinal tract.

Several factors are crucial for successful yogurt making:

 good sterile technique (i.e., proper sterilization and cooling of milk, proper cleansing and heat
treatment of glassware and 
 keeping out unwanted bacteria)
 proper incubation temperature (Lactobacilli and Streptococcus thermophilus are thermophilic
bacteria, meaning they prefer 
 elevated temperatures for growth. At such temperatures (50°C,
in this case) pathogenic or putrefactive bacteria are inhibited. However, even these
thermophilic bacteria are killed if exposed to temperatures over 55°C (130°F) and do not grow
well below 37°C (98°F))
 protection of the starter from contamination (Do not open the starter until you are ready to
make the next batch)

Materials:
1 liter of fresh milk (not non-fat milk)

Yogurt starter (Nestle plain yogurt)

disposable spoon

Transparent microwavable containers (small, around 100 - 250 ml, 1 per group)

Procedure:
1. Make sure that the microwavable container is clean. Clean it using detergent and water. Rinse
thoroughly.
2. Pour boiled water on the containers and allow the water to stay for 5 minutes. This will kill
most bacteria that may 
 contaminate the yogurt. Repeat this step for the disposable spoons.
3. Label the containers with laboratory section and group number.
4. Pour the milk on the cleaned containers and put inside the microwave for 2 minutes. Cool the
milk to temperatures 
 between 40 to 60°C.
5. Add a teaspoon of yogurt starter to the milk and stir. Set aside in a warm room for at least 7
hours until set. Do not disturb.
6. To know whether the yogurt has set, tilt slightly the container to see if the yogurt retains its
shape. If it does, place it in the 
 refrigerator. This may serve as starter culture for another
batch of yogurt. Do not open it unless the materials to make 
 another batch of yogurt are
ready.
7. To enjoy the yogurt, add sugar or salt. Yogurt could also be substituted for sour cream in dips
and other recipes.

Department of Biology, School of Science and Engineering

Ateneo de Manila University
Bi 6 Biotechnology for Everyone, Laboratory

Exercise 5. Fermentation

Name:___________________________________________ Date: _______________________

Group No.: _______________________________________ Instructor: ___________________

Guide Questions:
1. What are the health benefits of yogurt? Cite at least two original scientific papers that support
the health benefits of yogurt.
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2. Give a situation when your own body cells perform fermentation. Which body cells can
perform fermentation?
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3. It is said to be possible for lactose intolerant individuals to eat yogurt. Why?

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Department of Biology, School of Science and Engineering

Ateneo de Manila University