Production and Assay of Amylase by Bacillus Subtilis

Objectives
Extraction of commercially important enzymes from natural sources; development of enzyme assays;
quantification of enzyme activity and specific activity.
Bacillus Subtilis is known for amylase enzyme producer and starch or other types of sugar is commonly
used as the source of carbon during its fermentation. Therefore, this experiment is to extract the amylase
enzyme produced and to quantify the enzyme activity. In order to achieve the objectives of this
experiment, you have to:
1. Find the composition of fermentation medium containing starch to cultivate Bacillus Subtilis.
2. Find the best fermentation condition, i.e. temperature, shaking frequency, pH, and duration of
fermentation.
3. Find the method to extract the amylase enzyme.
4. Find the method to measure the enzyme activity.
5. Find the method to measure the specific activity.
Hint:
You need to prepare calibration curves. What is calibration curve and how is it used?

Questions:
Why starch is used in the fermentation medium?
What is the enzyme activity of amylase enzyme produced?