1. The Flavour of Vanilla; Satisfying Demand Through Bioconversion M. Douglas Baumann

2. Vanilla; Plain but Pricey A ubiquitous flavour in N. American and European (esp. French) food and drink Also added to make-up, perfume, pharmaceuticals and tobacco But vanilla is the world’s 2 nd most expensive spice!

3. Worldwide Demand for Vanilla Total demand for vanilla extract is ~2000 tonnes/a. Only 50 t/a. is natural vanilla Remaining demand is met by synthetic and pure vanillin $$$

4. The Original; Natural Vanilla Vanilla pods are harvested from Vanilla plantifolia, an orchid The orchid grows as a vine, taking ~3a. to reach maturity Most labour intensive commercial crop in the world 1,800 USD/kg

5. Synthetic Vanilla (Vanillin) The single most important component of natural vanilla is vanillin; Produced synthetically at large scale for 12 USD/kg from petroleum sources Must be labeled as artificial or synthetic vanilla

6. Pure Vanilla (Vanillin) Any vanillin that is obtained from natural, biological sources Pure vanilla is not natural vanilla; it lacks the 200+ other compounds found vanilla extract Highly valued by companies in N. America and Europe because pure vanilla is considered a natural, organic product Priced at 700 USD/kg

7. Vanillin Through Bioconversion… Common feedstock is lignin, a polymer that comprises 20- 30% of woody plants by weight Rice and sugar beet pulps are cheap sources of lignin that are readily degraded by enzymes (enzyme) (fungi) Lignin (polymer) (enzyme) Lignin (oligomer)

8. …requires Enzymes… Lignin depolymerase (Trametes cingulata) or Lignin peroxidase (Phaneracheate chrysporium) produce oligomers Enzymes liberate ferulic acid from lignin oligomers Ferulic acid esterase is isolated from Aspergillus niger A Trametes sp. Deligninified cellulose

9. …and Fungi! Ferulic acid is the most common feedstock in commercial processes (e.g. Rhodia Chimie) Final step is a whole cell bioconversion (e.g. Pycnoporus cinnabarinus) Vanillin is recovered as an extra-cellular product (~1g/L)

10. Final Packaging Crystalline vanillin is the final product Reconstituted in a 35% ethanol/water solution. Equivalent concentration of steeped vanilla beans is 100g/L, or less than 1%vol vanillin!