The invention relates to a method for producing biogas from lignocellulose-containing biomass, preferably from straw, and to a plant for carrying out said method.

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 invention relates to a method for the recombinant production of olivetolic acid (OA) in a host species selected from amoebozoa, based on a hybrid-gene or enzyme of polyketide synthase 37 (PKS37) in which the C-terminal type III PKS domain from an amoeba is replaced by an olivetol synthase (OLS) from a plant, and is expressed together with an olivetolic acid cyclase from a multi-gene expression vector. Further provided is a recombinant amoebozoa host species, and an improved method for producing Δ.sup.9-tetrahydrocannabinol (THC) or other cannabinoids.

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

The present invention relates to a genetically modified host cell producing a bibenzylic acid or a derivative thereof expressing a) one or more genes encoding a polyketide synthase (PKS); b) one or more genes encoding a polyketide cyclase (PKC); and c) one or more genes encoding a double bond reductase (DBR); and one or more genes encoding polypeptides selected from d) a tyrosine ammonia lyase polypeptide (TAL); e) a phenylalanine ammonia lyase polypeptide (PAL); f) a cinnamate 4-hydroxylase polypeptide (C4H); g) a cytochrome p450 reductase polypeptide (CPR); h) a 4-coumarate-CoA ligase polypeptide (4CL); and/or i) a non-catalytic chalcone isomerase type III or IV polypeptide (CHIL); wherein the at least one gene is heterologous to the host cell.

Provide are modified host cells that are engineered to decrese expression of a product to undetectable levels in the presence of an exogenous agent, and increase expression of the product in the presence of another exogenous agent. The modified yeast strainshost cells do not express detectable levels of a precursor or substrate used to make the product. The product can be a cannabinoid or precursor thereof, and the substrate can be hexanoate. Also provided are methods for making a product using the modified host cells. The modified host cell can be a yeast strain, such as S. cerevisiae.

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.

Nucleic acid molecules encoding polypeptides having polyketide synthase activity have been identified and characterized. Expression or over-expression of the nucleic acids alters levels of cannabinoid compounds in organisms. The polypeptides may be used in vivo or in vitro to produce cannabinoid compounds.

The compositions and methods of the disclosure can be used to produce a cannabinoid in a host cell, such as a yeast cell. For example, the disclosure features host cells (e.g., yeast cells) modified to express one or more enzymes of a cannabinoid biosynthetic pathway, such as an acyl activating enzyme (AAE), a tetraketide synthase (TKS), a cannabigerolic acid synthase (CBGaS), and/or an olivetolic acid cyclase (OAC).

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 (CBCA), 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), CBCA synthase (CBCAS), HMG-Co reductase (HMG1), and/or farnesyl 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.