Method and cell line for production of phytocannabinoids and phytocannabinoid analogues in yeast

A method of producing phytocannabinoids or phytocannabinoid analogues, the method comprising the steps of:

• (i) providing a yeast cell comprising a first polynucleotide comprising bases 523 to 9966 of SEQ ID NO:
• 38, coding for a polyketide synthase enzyme from D. discoideum comprising a mutation at G1516R (DiPKSG1516R) that reduces activity at an active site of a C-Met domain as compared to wild type DiPKS from D. discoideum, and a second polynucleotide coding for a cytosolic prenyltransferase enzyme, wherein:
• the polyketide synthase enzyme produces at least one precursor chemical from malonyl-CoA, the precursor chemical having structure I:
• wherein, on structure I, R1 is an alkyl group with a chain length of 1, 2, 3, 4, or 5 carbons, R2 is H, carboxyl, or methyl, and R3 is H, carboxyl, or methyl;
• the polyketide synthase enzyme is not native to C. sativa;
• and
• the cytosolic prenyltransferase enzyme prenylates the at least one precursor chemical, providing at least one species of phytocannabinoid or phytocannabinoid analogue;
• and
• (ii) propagating the yeast cell under suitable conditions to provide a yeast cell culture, thereby producing phytocannabinoids or phytocannabinoid analogues from the at least one precursor chemical.