Bioprocess Technology

1. E.coli colonies were seen on the plate.
2. The colonies were transferred into a shaker flask. (100ml of LB broth and Ampicillin).
3. Arabinose was added into the culture medium.
4. Parameters of the culture medium were adjusted for optimum cell growth.

Answers to further questions:

On media preparation:

Peptide and peptone(bacto-tryptone) - provide essential amino acid to growing bacteria.

Na+ ions (NaCl) - for transportation and maintining osmotic balance

Trace elements (Nitrogen, Sulphur, Magnesium) - provide a plethora of organism component helpful for bacteria growth.

Purpose of Ampicilin is to inhibits the growth of other bacteria thus preventing contamination

It is added only after autoclaving because Ampicilin will be inacivated at high temperature.

On equipment media:

Calibration of pH probe is to maintain the pH of media using buffers.

Hydrochloric is a strong acid. It dissociates in water affecting the optimum composition of the fermenter.

A peristaltic pump is a type of positive displacement pump used for pumping a variety of fluids. The fluids are pumped, just like the wavy-like motions of the oesophagus.

On seed preparation:

Arabinose is to provide the essential nutrients and carbon for bacterial to maintain the functions of the bacteria cells.

Gloves are worn when transferring E.coli into the flask containing LB medium, thus preventing contamination of any other bacteria found on hands. pGLO transformed E. coli is kept in the freezer to prevent any bacteria from multiplying. Bacteria cannot survive or multiply under extreme low temperature.

Step-wise scale-up is performed instead of transferring directly into the fermenter to ensure all the contaminants in the fermenter are being removed thus preventing contamination.