Abstract: Synthesis of enantiopure cis-?-irone from a renewable carbon source Disclosed herein are natural and synthetic enzymes capable of performing a method of producing cis-?-irone. The method comprises providing an enzyme capable of converting psi-ionone to cis-?-irone; and contacting the enzyme and psi-ionone under suitable conditions to produce cis-?-irone. The enzyme may be a methyltransferase from Streptomyces albireticuli (SaMT), a promiscuous bifunctional methyltransferase/cyclase (pMT1) enzyme from Streptomyces, or a modified pMT1 enzyme with at least one substitution. The enzymes allow the in-vivo and in-vitro production of cis-?-irone including the use of glucose as feedstock for the biotransformation into cis-?-irone.

Abstract: A method of increasing terpenoid production in a host cell that produces one or more terpenoids, comprising: a) providing a host cell that produces one or more terpenoids, said host cell comprising a vector comprising a polynucleotide sequence encoding a terpene synthase enzyme; b) modifying the vector to: i. introduce an inducible promoter operably linked to the polynucleotide sequence encoding the terpene synthase enzyme; and ii. introduce a polynucleotide sequence encoding a ribosomal binding site (RBS) that increases translation initiation rate of the terpenoid compared to a wild type ribosomal binding site; c) determining the dosage of an inducer capable of inducing the inducible promoter; d) culturing the host cell in a culture medium in the presence of the inducer at the dosage determined from step c); and e) isolating the terpenoid from the culture medium.

Abstract: A method of increasing terpenoid production in a host cell that produces one or more terpenoids, comprising: a) providing a host cell that produces one or more terpenoids, said host cell comprising a vector comprising a polynucleotide sequence encoding a terpene synthase enzyme; b) modifying the vector to: i. introduce an inducible promoter operably linked to the polynucleotide sequence encoding the terpene synthase enzyme; and ii. introduce a polynucleotide sequence encoding a ribosomal binding site (RBS) that increases translation initiation rate of the terpenoid compared to a wild type ribosomal binding site; c) determining the dosage of an inducer capable of inducing the inducible promoter; d) culturing the host cell in a culture medium in the presence of the inducer at the dosage determined from step c); and e) isolating the terpenoid from the culture medium.