Exemplary embodiments provided herein include genetically engineering microorganisms, such as yeast or bacteria, to produce cannabinoids by inserting genes that produce the appropriate enzymes for the metabolic production of a desired compound.

The application relates to recombinant polypeptides having berberine bridge enzyme (BBE) activity that are derived from microorganisms, such as Phytohabitans sufuscus, Streptomyces sp. AJS327, Streptomyces varsoviensis, Actinomadura pelletieri, Streptomyces flaveolus, Streptomyces sp. Ru71. or Streptomyces sp. CNH287, and the use of these recombinant polypeptides in compositions and methods for the oxidative cyclization of prenylated compounds, such as CBGA, in carrying out the biosynthesis, including cell-free biosynthesis, of cyclized cannabinoid compounds, such as CBCA, THCA, and CBDA.

Aspects of the disclosure relate to biosynthesis of cannabinoids and cannabinoid precursors in recombinant cells and in vitro.

This document relates to using bidirectional, multi-enzymatic scaffolds to biosynthesize cannabinoids in recombinant hosts.

Aspects of the disclosure relate to biosynthesis of cannabinoids and cannabinoid precursors in recombinant cells and in vitro. Described are host cells comprising heterologous polynucleotides encoding polyketide synthase (PKS) variants that are capable of producing more divarinol in the presence of butyrate.

The disclosure relates to a non-natural flavin-dependent oxidase comprising at least one amino acid variation as compared to a wild type flavin-dependent oxidase, wherein the non-natural flavin-dependent oxidase does not comprise a disulfide bond, and wherein the non-natural flavin-dependent oxidase is capable of oxidative cyclization of a prenylated aromatic compound into a cannabinoid. The disclosure also relates to a nucleic acid, an expression construct, and an engineered cell for making the non-natural flavin-dependent oxidase. Also provided are compositions comprising the non-natural flavin-dependent oxidase; isolated non-natural flavin-dependent oxidase and methods of making the same; cell extracts comprising the non-natural flavin-dependent oxidase; methods of making cannabinoids; and compositions comprising a cannabinoid and/or the non-natural flavin-dependent oxidase.

Strains of yeasts are provided containing the genes for the production of cannabinoids from fatty acids. The enzymes that mediate cannabinoid production are localized to the cytosol, peroxisome or different compartments within the secretory pathway (e.g., endoplasmic reticulum, Golgi, vacuole) to ensure efficient production. The engineered microorganisms produce cannabinoids in a controlled fermentation process.

This document relates to using bidirectional, multi-enzymatic scaffolds to biosynthesize cannabinoids in recombinant hosts.

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.

The present invention relates to polynucleotides for the generation of genetically modified plants, algae or plastids thereof that are capable of producing cannabinoids. In an aspect, the present invention also relates to methods of producing cannabinoids.