Optimized cannabinoid synthase polypeptides
AssigneeDemetrix, Inc.
InventorsAndrew Horwitz, Jeff Wong, Darren Platt, Jeff Ubersax
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Abstract
The present disclosure provides engineered variants of a cannabidiolic acid synthase (CBDAS) polypeptide comprising an amino acid sequence of SEQ ID NO:3 with one or more amino acid substitutions, nucleic acids comprising nucleotide sequences encoding said engineered variants, methods of making modified host cells comprising said nucleic acids, modified host cells expressing said engineered variants, methods of producing cannabinoids or cannabinoid derivatives, and methods of screening engineered variants of the cannabidiolic acid synthase (CBDAS) polypeptide.
Claims27
An engineered variant of a cannabidiolic acid synthase (CBDAS) polypeptide comprising an amino acid sequence of SEQ ID NO:3 with one or more amino acid substitutions, wherein said one or more amino acid substitutions occurs at an amino acid selected from the group consisting of C12, F17, F18, S20, R31, N33, P43, L49, K50, L51, Q55, N56, N57, L59, M61, S62, V63, S66, L71, S75, 197, L98, S100, V103, T109, Q124, V125, 1129, L132, S137, V149, W161, K165, E167, S170, L171, A172, Y175, C180, A181, H208, A235, A250, M256, K260, L268, H309, T310, F316, L326, G378, K389, E406, M412, L415, S428, L439, 1445, N466, Y499, N527, P538, R541, H542, R543, and H544, and
- wherein the amino acid sequence has at least 85% sequence identity to SEQ ID NO:3.
An engineered variant of a cannabidiolic acid synthase (CBDAS) polypeptide comprising the amino acid sequence of SEQ ID NO:3 with one or more amino acid substitutions, wherein the one amino acid substitutions are selected from the group consisting of C12F, F17M, F18T, F18W, S20G, R31Q, N33K, P43E, L49E, L49K, L49Q, K50T, L51I, Q55E, Q55P, N56E, N57D, N57E, L59E, M61H, M61S, M61W, S62N, S62Q, V63M, S66D, L71A, L71H, L71Q, S75D, S75E, 197V, L98V, S100A, V103A, V103F, T109V, Q124D, Q124E, Q124N, V125E, V125Q, I129V, L132M, S137G, H143D, V149I, W161K, W161R, W161Y, K165A, E167P, S170T, L171I, A172V, Y175F, C180A, A181V, N196Q, N196T, N196V, H208T, A235P, A250T, M256V, K260C, K260W, L268I, H309V, T310A, T310C, F316Y, L326I, G378T, G378S, K389E, E406K, M412Q, L415M, S428L, L439M, 1445M, N466D, K474S,-Y499M, Y499V, N527E, P538T, R541E, R541V, H542V, R543A, R543E, H544E, and H544D.
Dependencies are inferred from the claim text.