Benzylisoquinoline alkaloids (BIA) producing microbes, and methods of making and using the same

摘要

Aspects of the invention include host cells that are engineered to produce benzylisoquinoline alkaloids (BIAs). The host cells include heterologous coding sequences for a variety of enzymes involved in synthetic pathways from starting compounds to BIAs of the host cell. Also provided are methods of producing the BIAs of interest by culturing the host cells under culture conditions that promote expression of enzymes encoded by the heterologous coding sequences of the host cells. Aspects of the invention further include compositions, e.g., host cells, starting compounds and kits, etc., that find use in methods of the invention.

主权项

1. A method of preparing a benzylisoquinoline alkaloid product, the method comprising:
culturing an engineered microbial cell under conditions suitable for protein production, wherein the engineered microbial cell is an opioid-producing engineered microbial cell, wherein the engineered microbial cell comprises a plurality of heterologous coding sequences for encoding a plurality of enzymes within a pathway for producing the benzylisoquinoline alkaloid product, and wherein at least one heterologous coding sequence of the plurality of heterologous coding sequences encodes an enzyme that is within a pathway that converts a thebaine to the benzylisoquinoline alkaloid product wherein the engineered microbial cell is a yeast cell: adding a starting compound to the cell culture; and recovering the benzylisoquinoline alkaloid product from the cell culture, wherein the benzylisoquinoline alkaloid product is selected from the group consisting of a neopine, neomorphine, codeinone, codeine, morphine, morphinone, hydrocodone, dihydrocodeine, oxycodone, 14-hydroxycodeine, hydromorphone, and dihydromorphine.