IGEM:IMPERIAL/2009/Feedback & Debriefs/Feedback 23 7

Thursday 24th July 2009 - Presentation to Advisors

 * Applications:PKU
 * 2 possible enzymes: PAL and PAH.
 * Issues with feasability (0.6g of phenylamine can be degraded to 10mg of purified enzyme per litre of culture)
 * Another enzyme found that doesnt convert to tyrosine, instead to a non-toxic compound.
 * Phosphorilate enzymes, expresses phosphokinase in e.coli, making it more stable in the gut and protease resistant.
 * Advisor comments:
 * Initially unsure about the method.
 * Focus on the "big picture": First identify the aminotransferase enzymes, then cofactors,etc.
 * Drill down the amino acid synthesis pathway: (could include modelling exercise, find catalitic aborption rate: level of absorption and size)
 * Dipeptide/peptide/amino acid absorption issues. Find experiments.
 * Focus on a simple encapsulation system
 * Dont focus on too many diseases, more generic definition
 * Modularity is KEY


 * Encapsulation process:
 * Alg8 can be polymerised in vitro?
 * Minimal system: 1 polymer and then think of transport
 * 2 populations of cells? 1 will lyse, 1 won't
 * Advisor comments:
 * Have a hybrid system, express acetate and add Ca to alginate?
 * After cloning unsure that the system will express
 * Does E.coli have a system for transporting sugars out? (colonic acid and trealose found a knockout strain)
 * Got 1 gene in E.coli which is present(regulator)
 * Turn on the regulator: Turns on the pathway and start forming colonic acid: Negative outer layer and add Ca to it.
 * 2nd pathway sounds easier than alginate...
 * Try avoid blue light?
 * Make sure that we have literature to back up pathway 2 (colonaic acid).
 * How long does the process take?
 * Is there an electron microscopy study?
 * Bacteria remain alive?
 * How do we get rid of outer layer?
 * Stops producing mechanism for removal.
 * Biobric alginate pathway(1): Can E.coli export poly-mannuronic acid? Is there an easy way to make it?
 * Can we add Ca to poly-mannuronic acid?
 * Colonaic acid may nucleate Ca
 * Needs more research


 * Others
 * Timer: Link modules 1 and 2. Make it tunable?
 * What is feasable? PKU and cellulase - Need to research into PKU
 * Keep modules flexible: Need more background on PKU (peptide absorption)

Next meeting: 3:30pm on Friday 24th July