Provided herein are cannabinoid analogs, including halogenated cannabinoid analogs, hydroxylated cannabinoid analogs, deuterated cannabinoid analogs, and tritiated cannabinoid analogs. The cannabinoid analogs can be prepared by partial or total expression in modified host cells, such as recombinantly modified yeast cells, optionally in combination with chemical synthetic steps.

The present invention relates generally to production methods, enzymes and recombinant yeast strains for the biosynthesis of clinically important prenylated polyketides of the cannabinoid family. Using readily available starting materials, heterologous enzymes are used to direct cannabinoid biosynthesis in yeast.

In an aspect, the disclosure provides methods for making neurotransmitters in a host organism. The neurotransmitters can be cannabinoids and derivatives of cannabinoids. The host cells can be microalgae, fungi or other host cells. In a related aspect, the disclosure provides host cells engineered to have biochemical pathways for making neurotransmitters such as cannabinoids.

The invention provides engineered biosynthetic pathways that can be used to produce cannabinoids from fatty acids, recombinant microorganisms incorporating such pathways, methods of biosynthetically producing cannabinoids from fatty acids, and cannabinoids so produced.

Disclosed herein are microorganism and methods that can be used for the synthesis of cannabigerolic acid (CBGA) and cannabinoids. The methods disclosed can be used to produce CBGA, Δ.sup.9-tetrahydrocannabinolic acid (THCA), cannabidiolic acid (CBDA), cannabichromenic acid (CBGA), Δ.sup.9-tetrahydrocannabivarinic acid (THCVA), cannabidivarinic acid (CBDVA), cannabichromevarinic acid (CBCVA), Δ.sup.9-tetrahydrocannabinol (THC), cannabidiol (CBD), cannabichromene (CBC). Enzymes useful for the synthesis of CBGA and cannabinoids, include but are not limited to acyl activating enzyme (AAE1), polyketide synthase (PKS), olivetolic acid cyclase (OAC), prenyltransferase (PT), THCA synthase (THCAS), CBDA synthase (CBDAS), CBC A synthase (CBCAS), HMG-Co reductase (HMG1), and/or famesyl pyrophosphate synthetase (ERG20). The microorganisms can also have one or more genes disrupted, such as gene that that controls beta oxidation of long chain fatty acids.

Enzymes involved in cannabinoid biosynthesis are recombinantly expressed in a host cell. The host cell may be a prokaryote (e.g. Escherichia coli) or a eukaryote (e.g. Yarrowia lipolytica). The enzymes include a heterologous cannabigerolic acid synthase as well as additional enzymes involved in the biosynthesis of cannabinoid precursors such as geranyl diphosphate, olivetol, olivetolic acid, divarin and/or divarinic acid. Methods are provided for producing C5-cannabinoids and/or C3-cannabinoids by fermentation of the recombinant host cell. Alternatively, cannabinoids can be produced by biotransformation of cannabinoid precursors in recombinant cells or by disrupted recombinant cells.

The present disclosure provides genetically modified host cells that produce a cannabinoid, a cannabinoid derivative, a cannabinoid precursor, or a cannabinoid precursor derivative. The present disclosure provides methods of synthesizing a cannabinoid, a cannabinoid derivative, a cannabinoid precursor, or a cannabinoid precursor derivative.

The present disclosure relates to recombinant prenyltransferase enzymes with increased thermostability and activity and the use of these enzymes in compositions and methods for biosynthesis involving prenylation reactions, including compositions and methods for the preparation of cannabinoids.

A method is provided for biosynthetic production of cannabinoids in microorganisms from a carbon source precursor. This method describes the genetic modifications needed to engineer microorganisms to produce cannabinoids as well as a method for identifying and quantifying cannabinoids from fermentation broth. A system is also provided for tuning the method to produce different cannabinoids of interest by systematically modulating the enzymes encoded by the genetic modifications introduced in the microorganism.

Various aspects of this disclosure relate to compositions comprising (i) one or more decarboxylated cannabinoids and (ii) either (a) cellulose I, (b) nucleic acids that comprise one or more nucleotide sequences that encode a geranyl-pyrophosphate-olivetolic acid geranyltransferase, (c) protein that comprises one or more amino acid sequences that encode a geranyl-pyrophosphate-olivetolic acid geranyltransferase, (d) two of a, b, and c, or (e) each of a, b, and c.