Objectives:

1. To analyze the effect of different glucose concentration on the growth of the yeast.
2. To study the kinetic parameters of the system which include the specific growth and
the doubling time of the yeast.
Theory
Saccharomyces Cerevisiae which is also known as the bakers yeast commonly used as
the leavening yeast in the bakery.

Methodology
A. Preparation of the Yeast Agar Plate
1. First, turn on the laminar air flow cabinet and set up your materials in a sterile
environment such as a laminar flow hood with a clean bench top.
2. Aseptically transfer a very small amount of sample to the edge of the agar plate
3. Sterilize the inoculating loop with a Bunsen Burner and allow the inoculating loop
to cool down and then drag the loop gently through first quadrant and continue to
streak the second quadrant of the plate in he sig-zag manner.
4. Sterilize again the loop and let it cool and then loop through second quadrant to
third quadrant using zig-zag method and repeat the steps for the fourth quadrant.
5. Step 5: Incubate your plate based on the needs of the bacterial species you are
trying to isolate and check for your colonies 24 hours later.
B. Preparation of the Fermentation Medium
1. Weight 0.30 g of malt extract, 0.50 g of peptone for a working volume of 100ml.
2. Weight another 1.0 g of glucose monohydrate for a working volume of 100 ml
and dissolve them in a 250 Erlenmeyer flask with distilled water.
3. Set the pH of the prepared media according to the desired pH.
4. Then, cotton-plugged and covered the flasks with aluminium foil to avoid
contamination.
5. Sterile the media in batch autoclave at 121 C for 15 min.
6. After sterilization, let the media to cool down at room temperature before
inoculation.
7. Repeat steps 1 to step 7 by increasing the glucose monohydrate to 2g and 3g.
C. Preparation for the Inoculum Culture
1. The laminar cabinet is setting up and turn on and all the apparatus is in the sterile
environment.
2. The fermentation media is divide into the 100ml (preparation for inoculum) and
150ml(experiment).
3. Sterile the inoculation loop in the laminar cabinet using Bunsen-Burner.
4. Then, take 1 loop of the yeast (1loop=2 touch) from the agar plate and place into
100ml of the fermentation media in the Erlenmeyer Flask .
5. Repeat Step 2 and Step 3 until 3 loop of the yeast is put into the fermentation
media.
6. Then, incubate the mixture overnight at 33 degree celcius and 150rpm.
7. Repeat all step C for the different 4% and 6% glucose concentration.

D. I. Effect of Different Glucose Concentration On the Batch Yeast Fermentation.

1. The laminar cabinet is setting up and all the apparatus is in the sterile experiment.
2. Measure the fermentation media at 100ml and the inoculum for 10ml using
suitable measuring cylinder.
3. Then, cotton plugged and cover the flask with aluminium foil.
4. Then, incubate the flask at 33 degree celcius at 150rpm .
5. Repeat step 1 to Step 4 for the 4% and 6% glucose concentration.
II. Measure the Cell Concentration
1. Take a 2 ml sample at 1 hour interval and measure the cell concentration by
turbidity measurement atoptical density-600nm in a cuvette.
2. Replug and cover the flask after sampling for each interval time before putting
back in incubator shaker.
3. Repeat the step 1 for every one hour for six times.
Expected Result
Time
1%glucose conc

Refernces

Turbidity(600nm)
4%glucose conc

6%glucose con