Methods For Increasing Production Of Cannabinoids In Yeast Cells
The present invention is a method for the biosynthesis of hundreds of compounds, mainly found in the cannabis plant. The starting material for these compounds can be any biological compound that is used/produced in a biological organism from the sugar family starting materials or other low cost raw materials processed via enzymes or within organisms to give final products. These final products include, but are not limited to: cannabinoids, terpenoids, stilbenoids, flavonoids, phenolic amides, lignanamides, spermidine alkaloids, and phenylpropanoids. Specifically, the present invention relates to the regular, modified, or synthetic gene(s) of select enzymes that are processed and inserted into an expression system (for example, a vector, cosmid, BAC, YAC, phage) to produce modified hosts. The modified host is then optimized for efficient production and yield via manipulation, silencing, and amplifying inserted or other genes in the host, leading to an efficient system for product.
Claim 1. A method for increasing production of a cannabinoid or cannabinoid precursor molecule, comprising: providing a genetically modified yeast cell comprising: (i) a codonoptimized polynucleotide sequence expressing cannabinoid or cannabinoid precursor molecule producing enzymes, and (ii) a mutation to prevent or reduce endogenous sterol synthesis selected from the group consisting of: a SUE (sterol uptake exogenous)mutation and an ERG1 (Squalene monooxygenase) gene knockoutcontacting the genetically modified yeast cell with a starting materialand culturing the genetically modified yeast cell under conditions in which the cannabinoid or cannabinoid precursormolecule is produced from the starting material, wherein the cannabinoid or cannabinoid precursor molecule is selected from the group consisting of: cannabigerolic acid (CBGA), cannabigerovarinic acid (CBGVA), cannabidiolic acid (CBDA), cannabidivarinicacid (CBDVA), tetrahydrocannabinolic acid (THCA), delta-9-tetrahydrocannabivarinic acid (THCVA), cannabichromenic acid (CBCA), cannabichromevarinic acid (CBCVA), cannabigerol (CBG), cannabigerovarin (CBGV), cannabidiol (CBD), cannabidivarin (CBDV),tetrahydrocannabinol (THC), delta-9-tetrahydrocannabivarin (THCV), cannabichromene (CBC), cannabichromevarin (CBCV), olivetol, olivetolic acid, divarinic acid, divarinol, isopentenyl pyrophosphate (IPP), geranyl diphosphate (GPP) and farnesylpyrophosphate (FPP), and wherein the starting material is selected from the group consisting of: rice, soya, maize, wheat, beans, sugar beet, sugar cane, plant biomass, starch, cellulose, ethanol, lignocellulose, high fructose corn syrup, molasses, fattyacids, glycerol, lactic acid, whey and glucose.