Food Biotechnology - Bacillus Cultures - Biosurfactant Using Bacillus Subtilis - Essay Writing Biotechnology Assignment Help

Assignment Task:

AIM
The aim of these experiments is to use Bacillus cultures to produce biosurfactant in fermentation. Students will be provided with samples taken from different time points during the fermentation. 

Production of biosurfactant using Bacillus subtilis 

EQUIPMENT
Each group should have:
• 1 ml nutrient broth as control
• One piece of parafilm (roughly10 cm in length)
• 10 X empty Petri dishes
• 3 X universal tubes
• 3 X Cuvette
• 200 ml distilled water
• 200 ul crude oil
• 20 ul Triton X-100
• Access to a Gilson micropipette & tips (p1000 and p100)
• Access to centrifuge
• Access to spectrophotometer at wavelength 600 nm
• Access to pH metre (or sticks)

PROCEDURE
For all samples provided (No. 1 – 3 tubes), the following tests will be carried out (in triplicates):
1. Optical density @600nm: The OD600 (using nutrient broth as control to calibrate the spectrophotometer) using spectrophotometer. Follow the instructions available at the instrument.

2. The culture broth from the tubes will be centrifuged at 3000 rpm for 5 minutes. Decant the liquid (supernatant) in a tube and measure the pH.

3. Drop collapse assay: 35 ul of the supernatant will be pipetted as a drop onto parafilm, then the spreading of the drop on the parafilm surface will be measured after 15 minutes. Water will be used as the control.

4. Oil-spreading method: 20 ml distilled water will be added into Petri dishes, then 20 ul of crude oil will be added to the surface of the water. 10 ul of supernatant will then be added to the oil surface. If biosurfactant is present, the oil will be displaced with oil free clearing zone, and the diameter of this clearing zone will be recorded to indicate the surfactant activity. Triton X-100 was used as the positive control.

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