SynBERC:Drugs

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

Parts

• malic acid to malic semialdehyde converting enzyme

• 7-deoxyloganin synthetic enzymes (from glucose + GPP)

• vinblastine/vincristine transporter

Devices

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

Chassis

• Precursor supply
  • tryptophan
  • geranyl diphosphate (GPP)
  • malic acid

• Free glucose (PTS-independent) transport