Application US20200165641

BIDIRECTIONAL MULTI-ENZYMATIC SCAFFOLDS FOR BIOSYNTHESIZING CANNABINOIDS
[+] Show Details

US20200165641

1. A host cell capable of producing one or more cannabinoids selected from the group consisting of cannabigerolic acid, cannabidiolic acid, and cannabichromenic acid, said host cell comprising at least three different exogenous nucleic acids, (14) (2)

wherein said first and said second exogenous nucleic acids each encode a plurality of engineered enzymes selected from the group consisting of an acetyl-CoA acetyltransferase, a 3-hydroxybutyryl-CoA dehydrogenase, an enoyl-CoA hydratase, a beto-ketothiolase, a trans-enoyl-CoA reductase, an HMG-CoA synthase, an HMG-CoA reductase, a mevalonate kinase, a phosphomevalonate kinase, a diphosphomevalonate decarboxylase, an isopentenyl-diphosphate delta isomerase, a geranyl-diphosphate synthase, an olivetol synthase, an olivetolic acid cyclase, and a CBGA synthase; wherein each of said engineered enzymes comprises a heterologous interaction domain, said heterologous interaction domain comprising a first and a second peptide motif, and wherein each said heterologous interaction domain is different from each other; and

wherein said third exogenous nucleic acid encodes a polypeptide scaffold comprising a plurality of peptide ligands, wherein each said peptide ligand comprises an amino acid sequence that can bind to said first or said second peptide motif of one of said heterologous interaction domains.

2. The host cell of claim 1, wherein said plurality of engineered enzymes further comprises an ATP citrate lyase and an acetyl-CoA carboxylase. (0)

3. The host cell of claim 1, wherein said host cell further comprises an exogenous nucleic acid encoding a cannabidiolic acid synthase and a cannabichromenic acid synthase. (0)

4. The host cell of claim 1, wherein said host cell further comprises an exogenous cannabidiolic acid synthase. (0)

5. The host cell of claim 1, wherein said host cell further comprises an exogenous cannabichromenic acid synthase. (0)

6. The host cell of claim 1, wherein said host cell is a bacterial or a yeast host cell. (2) (0)

9. The host cell of claim 1, wherein said host cell is an algae or a plant cell. (2) (0)

12. The host cell of claim 1, said host cell comprising at least four different exogenous nucleic acids, wherein said first, said second, and said fourth nucleic acid each encode said plurality of engineered enzymes. (0)

13. The host cell of claim 1, said host cell comprising at least five different exogenous nucleic acids, wherein said first, said second, said fourth, and said fifth nucleic acid each encode said plurality of engineered enzymes. (0)

14. The host cell of claim 1, said host cell comprising at least six different exogenous nucleic acids, wherein said first, said second, said fourth, said fifth, and said sixth nucleic acid each encode said plurality of engineered enzymes. (0)

15. The host cell of claim 1, wherein each of said engineered enzymes is of the formula: enzyme-linker₁-spacer-linker₂-motif₁-linker₃-motif₂, where linkers 1, 2, and 3 can be the same or different, motif 1 and motif₂can be the same or different, and where motif 1 and motif 2 form said heterologous interaction domain. (3) (0)

18. The host cell of claim 1, said host cell further comprising a nucleic acid encoding a second polypeptide scaffold comprising a plurality of peptide ligands, wherein each said peptide ligand comprises an amino acid sequence that can that can bind to said first or said second peptide motif of one of said heterologous interaction domains. (0)

21. The host cell of claim 1, wherein each said exogenous nucleic acid comprises a constitutive promoter operably linked to the sequence encoding said plurality of engineered enzymes or said polypeptide scaffold. (2) (0)

23. The host cell of claim 1, wherein each said exogenous nucleic acid comprises an inducible promoter operably linked to the sequence encoding said plurality of engineered enzymes or said polypeptide scaffold. (0)

25. A method of producing one or more cannabinoids selected from the group consisting of cannabigerolic acid, cannabidiolic acid, and cannabichromenic acid, said method comprising culturing the host cell of claim 1 under conditions wherein said host cell produces said one or more cannabinoids. (4) (0)

View Abstract and Specification Size

PDF with Images and Document Face >

Full Text Publication >

Application US20200165641

BIDIRECTIONAL MULTI-ENZYMATIC SCAFFOLDS FOR BIOSYNTHESIZING CANNABINOIDS

Patent Matrix® Search