Abstract: 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.
Type:
Grant
Filed:
November 20, 2020
Date of Patent:
August 30, 2022
Assignee:
Hyasynth Biologicals Inc.
Inventors:
Timothy S. Liao, Letian Song, Louis Hom, Curtis Walton, Daniel Furlong, Mindy Melgar, Devanshi Bhargava
Abstract: A method and cell line for producing phytocannabinoids and phytocannabinoid analogues in yeast. The method applies, and the cell line includes, a yeast cell transformed with a polyketide synthase CDS and a cytosolic prenyltransferase CDS. The polyketide synthase enzyme catalyzes synthesis of olivetol or methyl-olivetol, and may include Cannabis sativa olivetolic acid synthase or Dictyostelium discoideum polyketide synthase (“DiPKS”). The yeast cell may be modified to include a phosphopantethienyl transferase for increased activity of DiPKS. The yeast cell may be modified to mitigate mitochondrial acetaldehyde catabolism for increasing malonyl-CoA available for synthesizing olivetol or methyl-olivetol. The prenyltransferase enzyme catalyzes synthesis of cannabigerol or a cannabigerol analogue, and may include an ???? cytosolic prenyltransferase enzyme from Streptomyces sp CL190.
Type:
Grant
Filed:
February 19, 2018
Date of Patent:
April 26, 2022
Assignee:
Hyasynth Biologicals Inc.
Inventors:
Shoham Mookerjee, Alexander James Campbell, Zachary Douglas Wiltshire, Kevin John Chen
Abstract: A method and cell line for producing polyketides in yeast. The method applies, and the cell line includes, a yeast cell transformed with a polyketide synthase coding sequence. The polyketide synthase enzyme catalyzes synthesis of olivetol or methyl-olivetol, and may include Dictyostelium discoideum polyketide synthase (“DiPKS”). Wild type DiPKS produces methyl-olivetol only. DiPKS may be modified to produce olivetol only or a mixture of both olivetol and methyl-olivetol. The yeast cell may be modified to include a phosphopantethienyl transferase for increased activity of DiPKS. The yeast cell may be modified to mitigate mitochondrial acetaldehyde catabolism for increasing malonyl-CoA available for synthesizing olivetol or methyl-olivetol.
Type:
Grant
Filed:
February 19, 2018
Date of Patent:
August 3, 2021
Assignee:
Hyasynth Biologicals Inc.
Inventors:
Shoham Mookerjee, Alexander James Campbell, Zachary Douglas Wiltshire, Kevin John Chen
Abstract: A method and cell line for producing polyketides in yeast. The method applies, and the cell line includes, a yeast cell transformed with a polyketide synthase coding sequence. The polyketide synthase enzyme catalyzes synthesis of olivetol or methyl-olivetol, and may include Dictyostelium discoideum polyketide synthase (“DiPKS”). Wild type DiPKS produces methyl-olivetol only. DiPKS may be modified to produce olivetol only or a mixture of both olivetol and methyl-olivetol. The yeast cell may be modified to include a phosphopantethienyl transferase for increased activity of DiPKS. The yeast cell may be modified to mitigate mitochondrial acetaldehyde catabolism for increasing malonyl-CoA available for synthesizing olivetol or methyl-olivetol.
Type:
Grant
Filed:
March 24, 2020
Date of Patent:
April 13, 2021
Assignee:
Hyasynth Biologicals Inc.
Inventors:
Shoham Mookerjee, Alexander James Campbell, Zachary Douglas Wiltshire, Kevin John Chen