1. Commercial Terpene Production Using Engineered Saccharomyces cerevisiae Bio-Based Chemicals East Boston, Massachusetts September 14, 2010 1
2. Accelerating Use of Industrial Biotechnology Will Transform the Chemical Market Advances in metabolic engineering are enabling the manufacture of diverse products that are impractical to produce synthetically Natural or sustainable replacements forexisting petrochemicals New sustainable chemicals with improvedor novel properties Rapid improvement in yields allows cost-effective production and broad use Sustainable biologic processes have lower environmental footprint than their petrochemical counterparts Reduction/elimination of toxic byproducts Reduced greenhouse gas emissions Abundant and renewable raw materials 2
5. Allylix Technology Proprietary metabolical engineering and fermentation platform for the production of rare and chemically complex terpenes Optimized production of bio-identical commercial compounds Development of novel compounds with valuable properties Advantages: Cost effective Sustainable supply Consistent quality Strong patent position 5
6. Allylix Platform Allows Biosynthetic Production of Commercially Inaccessible Sesquiterpenes Sesquiterpenes Flavors & Fragrances Urban Pesticide Crop Protection Food Ingredients/Nutraceuticals Cosmetic Compounds Pharmaceutical Intermediates Historically expensive to produce Multicyclic, multichiral compounds difficult and expensive to synthesize Low natural abundance makes them expensive to extract 6 Nootkatone
7. Allylix has Scaled and Commercialized the First Products From its Metabolic Engineering Platform 7 Built integrated technology platform and developed pipeline of products Scaled fermentation and purification processes to commercial scale Commenced commercial product and sales of first two products Advanced pipeline of six additional products that address multiple markets Flavor and fragrance Food ingredient/nutraceutical Crop protection/urban pesticides Cosmetic chemicals
8. 8 Sesquiterpene Production in Nature Glucose Manyenzymatic steps (ubiquitous) FPP Sesquiterpene synthase (plant specific) Valencene
10. Allylix Platform for Terpene Production Gene Isolation Synthesis, informatics, cloning Protein engineering Generating improved synthases Metabolic engineering Production of high levels of FPP for conversion to terpenes Fermentation Economical production of terpenes Combinatorial chemobiosynthesis Chemical modification of biosynthetic terpenes to produce novel or commercially-inaccessible products 10
11. Protein Engineering Used to Develop Terpene Synthases with Optimized Specificity and Catalytic Efficiency Altering specific amino acids in a terpene synthase to generate one with improved characteristics Improved synthases: Specificity and selectivity Optimize production of desired product Catalytic efficiency More active, robust enzymes 11
15. Allylix Fermentation Process Offers a Step Change in the Cost of Production 14 When fully developed: one commercial scale fermentation will produce as much valencene as 1.2 million metric tons of oranges
16. Discovery of and Screening of Novel Terpenes X X Chemical modification of biosynthetic terpene scaffolds to produce novel or commercially-inaccessible products “Smart” libraries built on chiral scaffolds more likely to produce compounds with desirable properties – “hits” Natural products produce higher proportion of hits than randomly generated products of combinatorial chemistry Natural products more closely resemble bioactive compounds Renewed emphasis on natural products screening and screening of semisynthetic natural product libraries Y Primary Derivatives (Dozens per scaffold) Secondary Derivatives (Hundreds per scaffold) Scaffolds (Several) 15
17. Summary The accelerating use of industrial biotechnology to produce renewable chemicals will transform the chemical market Allylix has scaled and commercialized the first products from its metabolic engineering platform Allylix platform technology supports a cost-effective and sustainable supply of commercial terpenes Modular platform allows development of new compounds by introduction new synthases into engineered yeast host strains with incremental investments of time, effort, and cost New terpenescan be and discovered and produced through generation and screening of chemobiosynthesiclibraries 16
