Recombinant polypeptides with berberine bridge enzyme activity useful for the biosynthesis of cannabinoids
AssigneeInvizyne Technologies, Inc.
InventorsTyler KORMAN, John BILLINGSLEY, Mohammad HAYAT
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Abstract
The application relates to recombinant polypeptides having berberine bridge enzyme (BBE) activity that are derived from microorganisms, such as Phytohabitans sufuscus, Streptomyces sp. AJS327, Streptomyces varsoviensis, Actinomadura pelletieri, Streptomyces flaveolus, Streptomyces sp. Ru71. or Streptomyces sp. CNH287, and the use of these recombinant polypeptides in compositions and methods for the oxidative cyclization of prenylated compounds, such as CBGA, in carrying out the biosynthesis, including cell-free biosynthesis, of cyclized cannabinoid compounds, such as CBCA, THCA, and CBDA.
Claims19
A method for preparing a compound of structural formula (I)
- wherein, R1 is C1-C7 alkyl,
- comprising contacting under suitable reactions conditions a compound of structural formula (II)
- wherein, R1 is C1-C7 alkyl,
- and a recombinant polypeptide comprising an amino acid sequence of at least 80%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to a sequence selected from SEQ ID NO:
- 8, 2, 4, 6, 10, 12, and 14.
A composition comprising:
- (a) a recombinant polypeptide having BBE activity comprising an amino acid sequence of at least 80%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to a sequence selected from SEQ ID NO:
- 8, 2, 4, 6, 10, 12, and 14;
- and (b) one or more enzymes that produce a substrate for the recombinant polypeptide.
A recombinant polypeptide having berberine bridge enzyme (BBE) activity comprising an amino acid sequence of at least 80% identity to SEQ ID NO:
- 2 and amino acid residue differences as compared to SEQ ID NO:
- 2 at one or more positions selected from T325, F156, L238, L283, T325, and G340;
- optionally, wherein the amino acid residue differences are selected from:
- T325G, F156Y, L238S, L283S, and G3401.
Dependencies are inferred from the claim text.