Methods of producing nor-opioid and nal-opioid benzylisoquinoline alkaloids
AssigneeAntheia, Inc.
InventorsChristina D. Smolke, Catherine Thodey, Isis Trenchard
Life of this patent
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Abstract
A method of demethylizing an opioid to a nor-opioid is provided. The method comprises contacting an opioid with at least one enzyme. Contacting the opioid with the at least one enzyme converts the opioid to a nor-opioid. A method of converting a nor-opioid to a nal-opioid is provided. The method comprises contacting a nor-opioid with at least one enzyme. Contacting the nor-opioid with the at least one enzyme converts the nor-opioid to a nal-opioid.
Claims18
-3. (canceled)
A method of altering an opioid to a nal-opioid, comprising:
- contacting the opioid with at least a first enzyme, wherein contacting the opioid with the at least a first enzyme converts the opioid to a nor-opioid through removal of a N-linked methyl group from the opioid;
- and
- contacting the nor-opioid with at least a second enzyme, wherein contacting the nor-opioid with the at least a second enzyme in the presence of a cofactor converts the nor-opioid to the nal-opioid through transfer of a sidechain from the cofactor.
A method of altering an opioid to a nal-opioid, comprising:
- contacting a first opioid with at least one enzyme, wherein contacting the first opioid with the at least one enzyme converts the first opioid to a second opioid through loss of an O-linked methyl group;
- contacting the second opioid with at least a second enzyme, wherein contacting the opioid with the at least a second enzyme converts the second opioid to the nor-opioid through loss of an N-linked methyl group;
- and
- contacting the nor-opioid with at least a third enzyme, wherein contacting the nor-opioid with the at least a third enzyme in the presence of a cofactor converts the nor-opioid to a nal-opioid through transfer of a sidechain from the cofactor.
-8. (canceled)
-12. (canceled)
-16. (canceled)
An engineered cell that produces a nor-opioid from an opioid present within the engineered cell, the engineered cell comprising a heterologous coding sequence encoding an N-demethylase produced by the engineered cell, wherein the N-demethylase converts the opioid within the engineered cell to the nor-opioid and wherein the nor-opioid is produced within the engineered cell.
-45. (canceled)
A method of altering a nor-opioid to a nal-opioid, comprising:
- contacting said nor-opioid with at least one enzyme, wherein contacting said nor-opioid with said enzyme in the presence of a cofactor converts said nor-opioid to a nal-opioid through transfer of a sidechain from the cofactor.
-64. (canceled)
A method for forming a product stream having a nor-opioid product, comprising:
- i. providing engineered cells and a feedstock including nutrients and water to a batch reactor, which engineered cells have at least one heterologous coding sequence comprising a N-demethylase enzyme;
- ii. in said batch reactor, subjecting said engineered cells to fermentation by incubating said engineered cells for a time period of at least about 5 minutes to produce a solution comprising said nor-opioid and cellular material;
- and
- iii. using at least one separation unit to separate said nor-opioid from said cellular material to provide said product stream comprising said nor-opioid.
-67. (canceled)
Dependencies are inferred from the claim text.