OBJECTIVES
Generating large quantities of a microbial product is not an easy task. We learnt that many considerations must be taken; type of bacteria, type of aeration, type of agitation, temperature, pH and monitoring processes amongst others. Care must be taken at every step as otherwise; the entire batch might be contaminated. Fermentation itself is quite useful to grow large quantities of cells and harvesting the product. Its ideas are simple, but putting it into practice is anything but easy.
Overall objective:
->To isolate green fluorescence protein (GFP) from E. coli
Experiment 1: Familiarisation with the Bioreactor and its Operation
->To get familiarized with the parts and components of microbial and mammalian bioreactors
->To be introduced to the basic operation procedure of a bioreactor
Experiment 2: Equipment, Media and Seed Culture Preparation
->To describe the steps to prepare a bioreactor
->To prepare the media for seed culture and scale-up fermentation
->To prepare seed culture for scale-up fermentation
Experiment 3: Inoculation, Fermentation and Monitoring
->To carry out scale-up fermentation to increase yield of desired protein product (GFP in this case)
->To monitor cell growth and product formation through manual sampling and computer data logging
Experiment 4: Isolation and Purification of Product
->To carry out sonication to lyse cells and isolate cell content from cell debris
->To carry out gel permeation chromatography to purify the GFP
Labels: Objectives