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Objective: Microbial synthesis of natural and unnatural organic compounds

This testbed is broken into two parts:

  1. Microbial synthesis of terpene indole alkaloids (TIAs), plant natural products
  2. de novo design of biosynthetic pathways


  • malic acid to malic semialdehyde converting enzyme
  • 7-deoxyloganin synthetic enzymes (from glucose + GPP)
  • vinblastine/vincristine transporter


Precursor Amplifiers

  • Over-expression of rate-limiting enzyme(s) for precursor formation
  • Simple expression device (promoter, RBS, ORF (protein coding), terminator)
  • Amp needs: tryptophan, geranyl diphoshpate (GPP), malic acid

Multi-cistronic/Multi-genic Biosynthetic Devices

  • Multiple ORFs with individual promoter/terminator pairs or as multi-cistronic operons

Metabolite Sensors

Needed to rapidly evaluate success in achieving synthesis goals.

  • secologanin
  • strictosidine
  • 3,4-dihydroxbutyric acid

Metabolite-controlled Biosynthetic Devices

  • Includes parts in multi-cistronic/multi-genic device
  • Metabolite-responsive expression (transcriptional control -- same switches can be used in sensors and control devices)
    • secolaganin
    • strictosidine
    • 3,4-dihydroxybutyric acid

Extracellular/Surface-associated Biosynthetic Device

  • Surface-anchoring protein part
  • promoter, RBS, ORF, terminator


  • Precursor supply
    • tryptophan
    • geranyl diphosphate (GPP)
    • malic acid
  • Free glucose (PTS-independent) transport