Part:BBa_K5443013

The Enoyl-CoA Hydratase/Aldolase (ECH) enzyme from Pseudomonas putida, ECH-Ppu, catalyses the fifth and final step in our vanillin biosynthesis pathway. ECH-Ppu converts feruloyl-CoA to vanillin by performing a two-step reaction: it hydrates the double bond of feruloyl-CoA, then cleaves the resulting thioester via a retro-aldol reaction (Fig. 1). The ECH enzyme can also potentially use caffeoyl-CoA and 4-coumaroyl-CoA as substrates.

We tested two alternate FCS enzymes for this pathway step; this enzyme from Pseudomonas, and also a homolog from Streptomyces (BBa_K5443014).

This part builds on the work of earlier iGEM teams, who contributed ECH Parts as follows: BBa_K1129002.

Figure 1. ECH-Ppu-catalysed conversion of feruloyl-CoA to vanillin.
This reaction represents the final step in our vanillin synthesis pathway. The ECH-Ppu enzyme converts feruloyl-CoA into vanillin.