Optimization of Protein Overproduction by Recombinant Microorganisms

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Three undergraduate students are to be supported by this supplementary request. Their resumes are included with this request. The proposed work involves three undergraduate research projects which are as follows: 1. Experimental Study of Plasmid Loss The undergraduate student will run continuous fermentations with bacteria containing plasmids encoded to overproduce a protein. Samples will be taken and analyzed in order to correlate plasmid loss with cell growth and protein production rate. Results will be compared with a mathematical model. 2. Modeling and Control of Invertase Overproduction The undergraduate student will perform shake flask experiments and scale-up experiments with a 10-15 liter fermentor in order to test and further develop a mathematical model of the secretion of overproduced invertase by yeast. 3. Selective Recycle via Flocculation and Sedimentation The undergraduate student will run a continuous recycle bioreactor configured to maintain a slower growing flocculent strain in spite of competition from a faster-growing non- flocculent strain. He or she will also perform coflocculation studies with mixed cultures and measure the degree of separation of the two strains that can be achieved by sedimentation in a inclined settler. Each of the three undergraduate students will be supervised by and meet at least biweekly with one of the co-PIs on the NSF grant. In addition, a graduate student supported by the NSF grant will be assigned as a mentor to each undergraduate student in order to assist the undergraduate with the day-to-day aspects of carrying out each project.