Title: 6.2 Lab: Running The Bioreactor
Course: ATEP - Biotechnology Module B > Section: Section 6 Test and Evaluate

6.2.1 Inoculating Your Bioreactor

A pure sample of your microorganism should be obtained from your instructor. The cells should be actively growing before putting them in the bioreactor so that they can readily metabolize the medium.

Generate an SOP to inoculate your bioreactor. Below is a sample of how inoculation may be performed. Use this as a starting point and expand upon it when relevant. You may need to return to your KSB charts from lesson 2.

1. Wear gloves.
2. Wipe down your bench using diluted bleach.
3. Pour distilled water into a flask.
4. Add medium ingredient to the distilled water. What are your medium ingredients? (See your KSBs.)
5. Bring solution to a boil. How will you bring it to a boil?
6. Test the pH of the solution. Add HCl until the pH you desire is reached. What is the desired pH? (See your KSBs.)
7. Boil on low for 15 minutes.
8. Cap the flask to cool slightly. With what will you cap the flask?
9. Sterilize a funnel. How can you sterilize the funnel?
10. Working quickly, remove the cap bottle, place the funnel in the bottle and pour the broth into the bioreactor.
11. Cap your bioreactor and begin aeration. This creates a positive pressure that makes it difficult for microbes to enter.
12. Open the cap again, insert the funnel and add the active microorganism. How much should you add?
13. Replace the cap and tighten.
14. Immediately perform an initial protease assay. How will you do this?

6.2.2 Maintaining Your Bioreactor and Collecting Data

Over the next few days, you will need to:

  1. Maintain optimal growth.
  2. Assay for protease.
  3. Check for contamination.
  4. Adjust variables, such as pH, aeration, oxygen levels and food sources as needed to maximize product.
  5. Refer to your SOPs as needed.

Be sure to document all data, observations and any SOP changes within your assignment.