Team:Rice University/STRATEGY

Pathway Design for Resveratrol Biosynthesis

TAL monomer.

• Tyrosine Ammonia Lyase (aka TAL, BBa_K122010) - TAL catalyzes the conversion of L-tyrosine to p-coumaric acid and ammonia. TAL also exhibits Phenylalanine Ammonia Lyase (PAL) activity, converting L-phenylalanine to trans-cinnamic acid and ammonia. Our work has focused on using Rhodotorula glutinis TAL because its ratio of TAL to PAL activity is high compared with other TAL homologs. In addition, previous studies have shown that this enzyme can be expressed as a functional protein in Saccharomyces cerevisiae and Escherichia coli. While the p-coumaric acid produced by TAL will serve as a substrate for resveratrol biosynthesis, the trans-cinnamic acid is expected to add a "floral" and "honey-like" bouquet to the beer.

Peanut STS monomer bound to resveratrol.

• 4-coumarate CoA ligase :: Stilbene Synthase Fusion Protein (aka 4CL:STS, BBa_K122005) - Completes the resveratrol biosynthetic pathway by catalyzing the conversion of p-coumaric acid to resveratrol, with a stoichiometry of 2:1. The 4CL:STS fusion protein was selected because it has been shown to more efficiently produce resveratrol than coexpression of the proteins separately (possibly due to substrate channeling).

Circuit Design

Cellular Chassis

Fermentation Apparatus