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US 11,447,788 B2Granted
System, methods, and compositions for high-level production of water-soluble cannabinoids in a plant cell culture
AssigneeTrait Biosceinces, Inc.
InventorsRichard T. Sayre, Elton Carvalho Gonçalves, Tawanda Zidenga
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Abstract
The inventive technology relates to systems and methods for enhanced in vivo production, accumulation and modification of cannabinoids. In one embodiment, the invention may include systems and methods for enhanced in vivo biosynthesis of chemically-modified water-soluble cannabinoids in a whole plant, or a cell suspension culture system.
Claims5
A method of glycosylating a cannabinoid comprising:
- 1. localizing one or more cannabinoid synthase enzymes in the cytosol of a Cannabis plant cell by expressing a heterologous nucleotide sequence, operably linked to a promoter, encoding:
- i. one or more of said cannabinoid synthase enzymes, wherein the N-terminal trichome targeting domain is disrupted or removed from the enzyme, and wherein said cannabinoid synthase is selected from the group consisting of:
- tetrahydrocannabinolic acid synthase (THCA synthase) according to SEQ ID NO:
- 46, or a sequence having at least 95% sequence identity with SEQ ID NO:
- 46, and cannabidiolic acid synthase (CBDA synthase) according to SEQ ID NO:
- 22, or a sequence having at least 95% sequence identity with SEQ ID NO:
- 22;
- ii. a heterologous catalase enzyme having reduction activity towards hydrogen peroxide generated by cannabinoid biosynthesis in the cytosol of said Cannabis cell;
- iii. a myb transcription factor, wherein said myb transcription factor is selected from the group consisting of:
- the nucleotide sequence of SEQ ID NO:
- 11, 12 and 43-45;
- iv. a heterologous glycosyltransferase selected from the group consisting of:
- SEQ ID NO:7, 24, 27, 29, 31, 33, 35, 37 and 39;
- and
- 2. biosynthesizing tetrahydrocannabinolic acid (THCA) and/or cannabidiolic acid (CBDA) in the cytosol of said Cannabis plant cell through the action of said THCA synthase and/or CBDA synthase, respectively.
12 more elements
An in vivo method for the high level production and accumulation of water-soluble cannabinoids comprising the steps of:
- 1. expressing in a Cannabis cell a nucleotide sequence, operably linked to a promoter, encoding:
- i. at least one cytosolic directed cannabinoid synthase enzyme having the trichome targeting domain disrupted or removed, wherein the cannabinoid synthase enzyme is selected from the group consisting of:
- SEQ ID NO:
- 22 and 46, or a sequence having at least 95% sequence identity with SEQ ID NO:
- 22 or 46;
- ii. a heterologous glycosyltransferase selected from the group consisting of:
- SEQ ID NO:
- 7, 24, 27, 29, 31, 33, 35, 37 and 39;
- iii. a catalase having reduction activity towards hydrogen peroxide generated by cannabinoid biosynthesis in the cytosol of said Cannabis cell;
- iv. a myb transcription factor, wherein said myb transcription factor is selected from the group consisting of:
- the nucleotide sequence of SEQ ID NO:
- 11, 12 and 43-45;
- and
- 2. glycosylating said at least one cannabinoid having at least one glycosylation site in the cytosol of said Cannabis cell forming at least one water-soluble cannabinoid glycoside, wherein said at least one cannabinoid is selected from the group consisting of delta-Δ9—tetrahydrocannabinol (THC);
- tetrahydrocannabinolic acid (THCA);
- cannabidiol (CBD);
- A cannabidiolic acid (CBDA);
- cannabigerol (CBG) cannabigerolic acid (CBGA).
15 more elements
Dependencies are inferred from the claim text.