1. 6/5/08Caltech iGEM Meeting #8 Presentation by Robert Ovadia Gut Microbes Lactose Intolerance

2. 6/5/08Caltech iGEM Meeting #8 Presentation by Robert Ovadia Introduction 2 From last week, we had three possible ways to tackle lactose intolerance: 1.Convert H2 or methane into less harmful gases 2.Make a super sugar metabolizing machine 3.Engineer a strain to NOT convert glucose + galactose into H2 and methane. Our Goals

3. 6/5/08Caltech iGEM Meeting #8 Presentation by Robert Ovadia Goal #2 3 Super Sugar Metabolizing Machine In order for our strain of E. coli to consume lactose, we must force it. In one of our earlier meetings, Doug spoke about his Healthbots, which included the phosphoenolpyruvate:sugar phosphotransferase system (PTS).

4. 6/5/08Caltech iGEM Meeting #8 Presentation by Robert Ovadia Goal #2 4 Super Sugar Metabolizing Machine To review, some bacteria have a prevelance for certain sugars, mainly glucose, fructose and sucrose (PTS sugars). To save energy, they will not consume non PTS sugars (lactose, maltose, arabinose), if PTS sugars are present (known as Carbon Catabolite Repression). We must toggle the PTS by inhibiting the blocking of LacY.

5. 6/5/08Caltech iGEM Meeting #8 Presentation by Robert Ovadia Goal #2 5 Super Sugar Metabolizing Machine

6. 6/5/08Caltech iGEM Meeting #8 Presentation by Robert Ovadia Goal #2 6 Super Sugar Metabolizing Machine To have our strain uptake lactose, we could introduce specific mutations in LacY. This will hopefully prevent the binding of EIIA(GLUC), allowing our strain to uptake lactose. In addition, we can overexpress LacY to help the process.

7. 6/5/08Caltech iGEM Meeting #8 Presentation by Robert Ovadia Goal #2 7 Super Sugar Metabolizing Machine

8. 6/5/08Caltech iGEM Meeting #8 Presentation by Robert Ovadia Reference 1.Hoischen C, Levin J, Pitaknarongphorn S, Reizer J, and Saier MH Jr. Involvement of the central loop of the lactose permease of Escherichia coli in its allosteric regulation by the glucose-specific enzyme IIA of the phosphoenolpyruvate-dependent phosphotransferase system. J Bacteriol 1996 Oct; 178(20) 6082-6. pmid:8830713. PubMed HubMed [Hoischen1996] 2.Deutscher, J., Francke, C. and Postma, P.W. 2006. How Phosphotransferase System-Related Protein Phosphorylation Regulates Carbohydrate Metabolism in Bactertia. Microbial and Molecular Biology Reviews. 2006: 939-1031