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Allylix
Allylix
Allylix
Allylix
Allylix
Allylix
Allylix
Allylix
Allylix
Allylix
Allylix
Allylix
Allylix
Allylix
Allylix
Allylix
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Allylix

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  • 1. Commercial Terpene Production Using Engineered Saccharomyces cerevisiae<br />Bio-Based Chemicals East<br />Boston, Massachusetts<br />September 14, 2010<br />1<br />
  • 2. Accelerating Use of Industrial Biotechnology Will Transform the Chemical Market <br />Advances in metabolic engineering are enabling the manufacture of diverse products that are impractical to produce synthetically<br />Natural or sustainable replacements forexisting petrochemicals<br />New sustainable chemicals with improvedor novel properties<br />Rapid improvement in yields allows cost-effective production and broad use<br />Sustainable biologic processes have lower environmental footprint than their petrochemical counterparts<br />Reduction/elimination of toxic byproducts<br />Reduced greenhouse gas emissions<br />Abundant and renewable raw materials<br />2<br />
  • 3. Industrial Biotechnology Market<br />3<br />Source: USDA<br />
  • 4. Synthetic Biology and Industrial Biotechnology<br />4<br />
  • 5. Allylix Technology<br />Proprietary metabolical engineering and fermentation platform for the production of rare and chemically complex terpenes<br />Optimized production of bio-identical commercial compounds<br />Development of novel compounds with valuable properties Advantages:<br />Cost effective<br />Sustainable supply<br />Consistent quality<br />Strong patent position <br />5<br />
  • 6. Allylix Platform Allows Biosynthetic Production of Commercially Inaccessible Sesquiterpenes<br />Sesquiterpenes<br />Flavors &amp; Fragrances<br />Urban Pesticide <br />Crop Protection<br />Food Ingredients/Nutraceuticals<br />Cosmetic Compounds<br />Pharmaceutical Intermediates<br />Historically expensive to produce<br />Multicyclic, multichiral compounds difficult and expensive to synthesize <br />Low natural abundance makes them expensive to extract<br />6<br />Nootkatone<br />
  • 7. Allylix has Scaled and Commercialized the First Products From its Metabolic Engineering Platform<br />7<br />Built integrated technology platform and developed pipeline of products<br />Scaled fermentation and purification processes to commercial scale<br />Commenced commercial product and sales of first two products<br />Advanced pipeline of six additional products that address multiple markets <br />Flavor and fragrance<br />Food ingredient/nutraceutical<br />Crop protection/urban pesticides<br />Cosmetic chemicals<br />
  • 8. 8<br />Sesquiterpene Production in Nature<br />Glucose<br />Manyenzymatic steps<br />(ubiquitous)<br />FPP<br />Sesquiterpene synthase<br />(plant specific)<br />Valencene<br />
  • 9. Sesquiterpene Synthase Diversity <br />9<br />
  • 10. Allylix Platform for Terpene Production<br />Gene Isolation<br />Synthesis, informatics, cloning<br />Protein engineering <br />Generating improved synthases<br />Metabolic engineering<br />Production of high levels of FPP for conversion to terpenes<br />Fermentation<br />Economical production of terpenes<br />Combinatorial chemobiosynthesis<br />Chemical modification of biosynthetic terpenes to produce novel or commercially-inaccessible products<br />10<br />
  • 11. Protein Engineering Used to Develop Terpene Synthases with Optimized Specificity and Catalytic Efficiency <br />Altering specific amino acids in a terpene synthase to generate one with improved characteristics<br />Improved synthases:<br />Specificity and selectivity<br />Optimize production of desired product<br />Catalytic efficiency<br />More active, robust enzymes<br />11<br />
  • 12. Metabolic Engineering for Cost-Effective Sesquiterpene Production by Fermentation<br />12<br />A<br />c<br />e<br />t<br />y<br />l<br />-<br />C<br />o<br />A<br />E<br />R<br />G<br />1<br />0<br />A<br />c<br />e<br />t<br />y<br />l<br />-<br />C<br />o<br />A<br />A<br />c<br />e<br />t<br />o<br />a<br />c<br />e<br />t<br />y<br />l<br />-<br />C<br />o<br />A<br />E<br />R<br />G<br />1<br />3<br />A<br />c<br />e<br />t<br />y<br />l<br />-<br />C<br />o<br />A<br />H<br />M<br />G<br />-<br />C<br />o<br />A<br />M<br />e<br />v<br />a<br />l<br />o<br />n<br />a<br />t<br />e<br />HMG1, HMG2<br />E<br />R<br />G<br />1<br />2<br />M<br />e<br />l<br />v<br />a<br />l<br />o<br />n<br />a<br />t<br />e<br />-<br />P<br />E<br />R<br />G<br />8<br />M<br />e<br />v<br />a<br />l<br />o<br />n<br />a<br />t<br />e<br />-<br />P<br />P<br />E<br />R<br />G<br />1<br />9<br />D<br />M<br />A<br />P<br />P<br />I<br />P<br />P<br />I<br />D<br />I<br />G<br />P<br />P<br />(<br />F<br />P<br />S<br />)<br />E<br />R<br />G<br />2<br />0<br />I<br />P<br />P<br />E<br />r<br />g<br />o<br />s<br />t<br />e<br />r<br />o<br />l<br />S<br />q<br />u<br />a<br />l<br />e<br />n<br />e<br />F<br />P<br />P<br />T<br />e<br />r<br />p<br />e<br />n<br />e<br />S<br />y<br />n<br />t<br />h<br />a<br />s<br />e<br />Eliminate bottlenecks in carbon flow to terpenes through:<br /><ul><li>Improved carbon flow from acetyl-CoA to FPP
  • 13. Modulation of squalenesynthase activity
  • 14. High level expression of optimized terpenesynthase</li></li></ul><li>Commercial Production<br />Fermentation and recovery of first products at commercial scale<br />Valencene: Orange flavor and fragrance<br />Nootkatone: Grapefruit flavor and fragrance<br />13<br />
  • 15. Allylix Fermentation Process Offers a Step Change in the Cost of Production <br />14<br />When fully developed:<br />one commercial scale fermentation<br />will produce as much valencene as<br />1.2 million metric tons of oranges<br />
  • 16. Discovery of and Screening of Novel Terpenes<br />X<br />X<br />Chemical modification of biosynthetic terpene scaffolds to produce novel or commercially-inaccessible products<br />“Smart” libraries built on chiral scaffolds more likely to produce compounds with desirable properties – “hits”<br />Natural products produce higher proportion of hits than randomly generated products of combinatorial chemistry<br />Natural products more closely resemble bioactive compounds<br />Renewed emphasis on natural products screening and screening of semisynthetic natural product libraries<br />Y<br />Primary Derivatives<br />(Dozens per scaffold)<br />Secondary Derivatives<br />(Hundreds per scaffold)<br />Scaffolds<br />(Several)<br />15<br />
  • 17. Summary<br />The accelerating use of industrial biotechnology to produce renewable chemicals will transform the chemical market <br />Allylix has scaled and commercialized the first products from its metabolic engineering platform<br />Allylix platform technology supports a cost-effective and sustainable supply of commercial terpenes<br />Modular platform allows development of new compounds by introduction new synthases into engineered yeast host strains with incremental investments of time, effort, and cost<br />New terpenescan be and discovered and produced through generation and screening of chemobiosynthesiclibraries<br />16<br />

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