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  • Is well-studied and well-characterized; lots of past work in E. coli
  • Past iGEM teams
    • Nevada 2011: "A Cooperative Relationship Between Cyanobacteria and E.Coli for Production of Biofuels"
    • WISE 2011
  • difference between diesel and gasoline
    • diesel freezes at a lower temperature than gasoline (used in Canadian cars)
    • diesel doesn't need a spark; just need compression
    • making gasoline is more complicated; more complicated mixture
    • diesel is more efficient, but less powerful 
  • What is biodiesel:
    • main component is sulfur ester
    • fatty acid methyl esters (FAME)
    • fatty acid ethyl esters (FAEE)
    • pathway:
      • glycolysis -> pyruvate -> acetyl coA -> Fatty acid synthesis -> fatty acid esters
      • glycolysis -> pyruvate -> acetaldehyde -> ethanol
  • How to get biodiesel out of cell?
    • need to lyse cells?
    • engineer bacteria to excrete it and then skim it off the top of the reactor? -E. coli exporters
  • Bioenergy landscape
  • Which genes? Where do we get them?
    • Are they in the iGEM kit?
  • How to measure?
    • "The fatty acid and ethanol assays were quantitated by generating standard curves of absorbance vs substrate concentration using known quantities of palmitic acid or ethanol standards. "

...

  • Group 1: cellulose -> intermediate via ?
    • growth conditions
    • measure byproducts and intermediate metabolites
    • growth dynamics
    • effects of pH, temperature, feed stock, inhibitory byproducts
    • Ubq ligase + Ab -> knockdown
    • genetic manipulations (e.g. can we express GFP?)
  • Group 2: intermediate -> biodiesel via E. coli
    • effects of pH, temperature, intermediates from bacteria #1, inhibitory byproducts
    • measure biodiesel (make an assay?)
    • Biodiesel transporter: measure internal and external FAEE
    • Which genes turn intermediates into biodiesel? construction of plasmids? regulation of genes?

Need to know for next week:

  • Which intermediates? Which pathways of cellulose production?