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