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 present disclosure relates generally to methods, isolated polypeptides and polynucleotides, expression vectors, and host cells for the production of olivetolic acid and phytocannabinoids. A method of producing olivetolic acid (OVLa) and/or a phytocannabinoid in a heterologous host cell having OVLa-producing or phytocannabinoid-producing capacity comprises transforming the host cell with a nucleotide encoding a variant olivetolic acid cyclase (OAC) protein having at least 6 amino acid mutations relative to the wild type OAC protein, and culturing the transformed host cell to produce OVLa and/or phytocannabinoids therefrom. The variant OAC protein (SEQ ID NO:92) has at least 85% sequence identity with the wild type OAC protein (SEQ ID NO:91). Exemplary variants having improved OVLa or phytocannabinoid production capacity are described.

The present disclosure relates to the production of cannabinoids in yeast. In as aspect there is provided a genetically modified yeast comprising: one or more GPP producing genes and optionally, one or more GPP pathway genes; two or more olivetolic acid producing genes; one or more cannabinoid precursor or cannabinoid producing genes; one or more Hexanoyl-CoA producing genes, and at least 5% dry weight of fatty acids or fats.

An expression system and method for producing a cannabinoid in algae are provided. The method includes expressing in an algae cell an enzyme for converting hexanoic acid to hexanoyl-CoA, enzymes for converting hexanoyl-CoA to olivetolic acid (OA), an enzyme for converting olivetolic acid (OA) to cannabigerolic acid (CbGA) and an enzyme for converting cannabigerolic acid (CbGA) to a cannabinoid.

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.

The present disclosure provides recombinant host cells comprising a pathway capable of producing a cannabinoid and/or cannabinoid precursor, wherein the pathway comprises an enzyme AAE from a source organism other than Cannabis sativa, such as Humulus lupulus. The disclosure also provides methods of using the host cells to produce rare cannabinoids and/or rare cannabinoid precursors.

Described herein are non-natural olivetol synthase (OLS) variants, nucleic acids, engineered cells, method s for preparing cannabinoids, and compositions thereof. The non-natural olivetol OLS variants form desired cannabinoid precursor and products at increased rates, have higher affinity for pathway substrates, and/or byproducts are formed in lower amounts in their presence, as compared to wild type OLS. The OLS variants can be used to form linear polyketides, and can be expressed in an engineered cell having a pathway to form cannabinoids, which include CBGA, its analogs and derivatives. CBGA can be used for the preparation of cannabigerol (CBG), which can be used in therapeutic compositions.