CPC C12P 17/12 (2013.01) [C12N 9/0006 (2013.01); C12N 9/0071 (2013.01); C12N 15/52 (2013.01); C12N 15/62 (2013.01); C12P 41/002 (2013.01); C12Y 101/01247 (2013.01); C12N 2800/22 (2013.01)] | 24 Claims |
1. 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.
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