Methods of producing epimerases and benzylisoquinoline alkaloids
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsChristina D. Smolke, Stephanie Galanie, Isis Trenchard, Catherine Thodey, Yanran Li
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Abstract
A method of epimerizing an (S)-1-benzylisoquinoline alkaloid to an (R)-1-benzylisoquinoline alkaloid is provided. The method comprises contacting the (S)-1-benzylisoquinoline alkaloid with at least one enzyme. Contacting the (S)-1-benzylisoquinoline alkaloid with the at least one enzyme converts the (S)-1-benzylisoquinoline alkaloid to an (R)-1-benzylisoquinoline alkaloid.
Claims24
A method of producing an (R)-1-benzylisoquinoline alkaloid, the method comprising:
- (a) culturing an engineered non-plant cell in a cell culture comprising a feedstock, wherein the engineered non-plant cell stably expresses a plurality of heterologous enzymes within the same engineered non-plant-cell for the production of an (S)-1-benzylisoquinoline alkaloid wherein the plurality of heterologous enzymes comprise tyrosine hydroxylase (TyrH), L-DOPA decarboxylase (DODC), norcoclaurine synthase (NCS), norcoclaurine 6-O-methyltransferase (6OMT), coclaurine N-methyltransferase (CNMT), cytochrome P450 80B1 (CYP80B1), and 4′-O-methyltransferase (4′OMT),
- (b) producing an (S)-1-benzylisoquinoline alkaloid within the same engineered non-plant cell,
- (c) contacting, within the same engineered non-plant cell, the (S)-1-benzylisoquinoline alkaloid that is produced within the engineered non-plant cell with at least one heterologous, fused enzyme expressed within the same engineered non-plant cell that converts the (S)-1-benzylisoquinoline alkaloid to an (R)-1-benzylisoquinoline alkaloid within the engineered non-plant cell, and
- (d) producing the (R)-1-benzylisoquinoline alkaloid within the engineered non-plant cell.
A method of producing an (R)-1-benzylisoquinoline alkaloid, the method comprising:
- (a) culturing an engineered non-plant cell in a cell culture comprising a feedstock, wherein the engineered non-plant cell stably expresses a plurality of heterologous enzymes within the same engineered non-plant-cell for the production of an (S)-1-benzylisoquinoline alkaloid wherein the plurality of heterologous enzymes comprise tyrosine hydroxylase (TyrH), L-DOPA decarboxylase (DODC), monoamine oxidase (MAO), norcoclaurine synthase (NCS), norcoclaurine 6-O-methyltransferase (6OMT), coclaurine N-methyltransferase (CNMT), and 4′-O-methyltransferase (4′OMT),
- (b) producing an (S)-1-benzylisoquinoline alkaloid within the same engineered non-plant cell,
- (c) contacting, within the same engineered non-plant cell, the (S)-1-benzylisoquinoline alkaloid that is produced within the engineered non-plant cell with at least one heterologous, fused enzyme expressed within the same engineered non-plant cell that converts the (S)-1-benzylisoquinoline alkaloid to an (R)-1-benzylisoquinoline alkaloid within the engineered non-plant cell, and
- (d) producing the (R)-1-benzylisoquinoline alkaloid within the engineered non-plant cell.
Dependencies are inferred from the claim text.