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

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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