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.
Claims21
A method of epimerizing an (S)-1-benzylisoquinoline alkaloid to an (R)-1-benzylisoquinoline alkaloid, comprising:
- (a) culturing an engineered non-plant cell, wherein the engineered non-plant cell stably expresses one or more heterologous enzymes within the same engineered non-plant-cell for the production of an (S)-1-benzylisoquinoline alkaloid wherein the one or more heterologous enzymes are selected from the group consisting of tyrosine hydroxylase (TyrH), L-DOPA decarboxylase (DODC), monoamine oxidase (MAO), norcoclaurine synthase (NCS), norcoclaurine 6-O-methyltransferase (6OMT), coclaurine N-methyltransferase (CNMT), N-methylcoclaurine 3′-hydroxylase (CYP80B1), and 4′-O-methyltransferase (4′OMT);
- and
- (b) 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 enzyme,
- wherein contacting the (S)-1-benzylisoquinoline alkaloid with the at least one enzyme converts the (S)-1-benzylisoquinoline alkaloid to an (R)-1-benzylisoquinoline alkaloid.
An engineered non-plant cell comprising:
- i) at least one enzyme that converts an (S)-1-benzylisoquinoline alkaloid to an (R)-1-benzylisoquinoline alkaloid;
- and
- ii) one or more stably expressed heterologous enzymes for the production of the (S)-1-benzylisoquinoline alkaloid within the engineered non-plant cell wherein the one or more heterologous enzymes are selected from the group consisting of tyrosine hydroxylase (TyrH), L-DOPA decarboxylase (DODC), monoamine oxidase (MAO), norcoclaurine synthase (NCS), norcoclaurine 6-O-methyltransferase (6OMT), coclaurine N-methyltransferase (CNMT), N-methylcoclaurine 3′-hydroxylase (CYP80B1), and 4′-O-methyltransferase (4′OMT),
- wherein an (S)-1-benzylisoquinoline alkaloid is epimerized to an (R)-1-benzylisoquinoline within the engineered non-plant cell.
Dependencies are inferred from the claim text.