Provided is an enzyme useful for prenylation and recombinant pathways for the production of cannabinoids, cannabinoid precursors and other prenylated chemicals in a cell free system as well and recombinant microorganisms that catalyze the reactions.

Provided is an enzyme useful for prenylation and recombinant pathways for the production of cannabinoids, cannabinoid precursors and other prenylated chemicals in a cell free system as well and recombinant microorganisms that catalyze the reactions.

This disclosure describes recombinant cells and methods for microbial biosynthesis of ent-atiserenoic acid. Thus, in one aspect, this disclosure describes a recombinant cell genetically modified to exhibit increased biosynthesis of ent-atiserenoic acid compared to a comparable control cell. In some cases, the recombinant cell can include a host cell modified to include at least one heterologous polynucleotide encoding at least one enzyme in a biosynthetic pathway that produces ent-atiserenoic acid. In some cases, the recombinant cell can include a host cell and at least one heterologous enzyme in a biosynthetic pathway that produces ent-atiserenoic acid.

Provided is an enzyme useful for prenylation and recombinant pathways for the production of cannabinoids, cannabinoid precursors and other prenylated chemicals in a cell free system as well and recombinant microorganisms that catalyze the reactions.

Provided is an enzyme useful for prenylation and recombinant pathways for the production of cannabinoids, cannabinoid precursors and other prenylated chemicals in a cell free system as well and recombinant microorganisms that catalyze the reactions.

This disclosure describes recombinant cells and methods for microbial biosynthesis of ent-atiserenoic acid. Thus, in one aspect, this disclosure describes a recombinant cell genetically modified to exhibit increased biosynthesis of ent-atiserenoic acid compared to a comparable control cell. In some cases, the recombinant cell can include a host cell modified to include at least one heterologous polynucleotide encoding at least one enzyme in a biosynthetic pathway that produces ent-atiserenoic acid. In some cases, the recombinant cell can include a host cell and at least one heterologous enzyme in a biosynthetic pathway that produces ent-atiserenoic acid.

A recombinant cell of Saccharomyces cerevisiae that includes in its genome nucleic acids encoding cannabinoid biosynthetic pathway genes. A cannabinoid is produced by the recombinant cell in the presence of a cannabinoid precursor substrate and at least one of the cannabinoid biosynthetic pathway genes is from an organism other than Cannabis sativa, wherein the at least one of the cannabinoid biosynthetic pathway genes encodes a prenyltransferase. In an embodiment, the prenyltransferase is NphB from Streptomyces sp. having the amino acid sequence of any one of SEQ ID NOs: 8-11. Also disclosed is a method for producing a cannabinoid with the recombinant cell and the cannabinoid precursor substrate.

A method for producing a cannabinoid precursor by contacting a substrate and geranyl pyrophosphate or farnesyl pyrophosphate with an NphB orthologue. The NphB orthologue is from an organism other than Cannabis sativa, and the substrate can be 2,4-dihydroxy-6-pentylbenzoic acid or 2,4-dihydroxy-6-propylbenzoic acid. Also disclosed is a recombinant cell of Yarrowia lipolytica, carrying in its genome a nucleic acid encoding an NphB orthologue from an organism other than Cannabis sativa such that the NphB orthologue is expressed in the recombinant cell.

Ways of making and using a recombinant organism configured to produce a cannabinoid are provided. The recombinant organism can include a eukaryotic microorganism expressing a recombinant construct including a geranyl diphosphate synthase (GPPS2), an isopentenyl diphosphate isomerase (IDI), an isopentenyl phosphate kinase (IPK), and a 5-(hydroxyethyl)-methyl thiazole kinase (ThiM). A cannabinoid can be produced by a process that includes growing the recombinant organism configured to produce the cannabinoid in a growth medium and separating the cannabinoid from the recombinant organism and the growth medium. A biosynthetic system for producing a cannabinoid is provided that includes a bioreactor, the recombinant organism configured to produce the cannabinoid, and a growth medium for the recombinant organism.

Provided is an enzyme useful for prenylation and recombinant pathways for the production of cannabinoids, cannabinoid precursors and other prenylated chemicals in a cell free system as well and recombinant microorganisms that catalyze the reactions.