The present disclosure provides methods and compositions for producing rotundone. In various aspects, the present disclosure provides enzymes, polynucleotides encoding said enzymes, and recombinant microbial host cells (or microbial host strains) for the production of rotundone. In some embodiments, the present disclosure provides microbial host cells for producing rotundone at high purity and/or yield, from either enzymatic transformation of α-guaiene, or from sugar or other carbon source. The present disclosure further provides methods of making products containing rotundone, including flavor or fragrance products, among others.

The present disclosure provides methods and compositions for producing rotundone. In various aspects, the present disclosure provides enzymes, polynucleotides encoding said enzymes, and recombinant microbial host cells (or microbial host strains) for the production of rotundone. In some embodiments, the present disclosure provides microbial host cells for producing rotundone at high purity and/or yield, from either enzymatic transformation of α-guaiene, or from sugar or other carbon source. The present disclosure further provides methods of making products containing rotundone, including flavor or fragrance products, among others.

The present disclosure provides methods and compositions for producing rotundone. In various aspects, the present disclosure provides enzymes, polynucleotides encoding said enzymes, and recombinant microbial host cells (or microbial host strains) for the production of rotundone. In some embodiments, the present disclosure provides microbial host cells for producing rotundone at high purity and/or yield, from either enzymatic transformation of α-guaiene, or from sugar or other carbon source. The present disclosure further provides methods of making products containing rotundone, including flavor or fragrance products, among others.

Described herein are chimeric terpene synthases, methods for making chimeric terpene synthases, and methods for making terpenes using the same.

The present disclosure in various aspects provides engineered enzymes and encoding polynucleotides, as well as host cells and methods for making rotundone and other terpenoids. For example, in various aspects, the invention provides engineered ?-Guaiene Synthase (?GS) and Guaiene Oxidase (GO) enzymes that increase biosynthesis of rotundone from farnesyl diphosphate, and in certain embodiments substantially reduce biosynthesis of side products such as ?-Bulnesene or oxygenated side products. In still other aspects, the invention provides engineered terpene synthase enzymes for directing biosynthesis toward a desired product, to thereby improve product profiles and/or product titers from terpene synthase reactions.

Described herein are chimeric terpene synthases, methods for making chimeric terpene synthases, and methods for making terpenes using the same.

The present disclosure provides methods and compositions for producing rotundone. In various aspects, the present disclosure provides enzymes, polynucleotides encoding said enzymes, and recombinant microbial host cells (or microbial host strains) for the production of rotundone. In some embodiments, the present disclosure provides microbial host cells for producing rotundone at high purity and/or yield, from either enzymatic transformation of -guaiene, or from sugar or other carbon source. The present disclosure further provides methods of making products containing rotundone, including flavor or fragrance products, among others.

The current disclosure relates to compositions and methods relating to engineered polypeptides that can encode guaiene synthases that are selective for alpha-guaiene. The engineered polypeptides can also catalyze the production of increased amounts of alpha-guaiene. The disclosure also relates to host cells including the engineered polypeptides that can be used to produce large quantities of alpha-guaiene and its derivatives. The disclosure also relates to methods of producing alpha-guaiene employing said host cells.