METHODS OF IMPROVING PRODUCTION OF MORPHINAN ALKALOIDS AND DERIVATIVES
Methods and systems are provided for producing codeinone within an engineered non-plant cell. The method comprises, within the engineered non-plant cell, producing a thebaine product. The method also comprises, within the engineered non-plant cell, contacting the thebaine product with an enzyme having thebaine 6-O-demethylase activity, thereby producing a neopinone product. Additionally, the method comprises, within the engineered non-plant cell, contacting the neopinone product with a neopinone isomerase, thereby producing a codeinone product.
1 . A method of producing codeinone within an engineered non-plant cell, the method comprising:
within the engineered non-plant cell, producing a thebaine product;
within the engineered non-plant cell, contacting the thebaine product with an enzyme having thebaine 6-O-demethylase activity, thereby producing a neopinone product; and
within the engineered non-plant cell, contacting the neopinone product with a neopinone isomerase, thereby producing a codeinone product.
2 . The method of claim 1 , wherein the neopinone isomerase is an engineered neopinone isomerase.
3 . The method of claim 1 , further comprising:
within the engineered non-plant cell, contacting the codeinone product with an enzyme having morphinone reductase activity, thereby producing a hydrocodone product.
4 . The method of claim 1 , further comprising:
within the engineered non-plant cell, contacting the codeinone product with an enzyme having codeinone reductase activity, thereby producing a codeine product.
5 . The method of claim 1 , further comprising:
within the engineered non-plant cell, contacting the codeinone product with an enzyme having morphine dehydrogenase activity, thereby producing a codeine product.
6 . A method of isomerizing neopinone within an engineered non-plant cell, the method comprising:
contacting a neopinone product with a neopinone isomerase,
wherein contacting the neopinone product with the neopinone isomerase isomerizes the neopinone product to a codeinone product, and
wherein the neopinone isomerase is produced by culturing an engineered non-plant cell having at least one coding sequence, encoded within a chromosome within the engineered non-plant cell, for encoding the neopinone isomerase.
7 . The method of claim 6 , wherein the neopinone isomerase is an engineered neopinone isomerase.
8 . The method of claim 6 , further comprising:
within the engineered non-plant cell, contacting the codeinone product with an enzyme having morphinone reductase activity, thereby producing a hydrocodone product.
9 . The method of claim 6 , further comprising:
within the engineered non-plant cell, contacting the codeinone product with an enzyme having codeinone reductase activity, thereby producing a codeine product.
10 . The method of claim 6 , further comprising:
within the engineered non-plant cell, contacting the codeinone product with an enzyme having morphine dehydrogenase activity, thereby producing a codeine product.
11 . The method of claim 1 , wherein at least one of products selected from the group consisting of the thebaine product and the neopinone product is produced within the engineered non-plant cell from a simple starting material.
12 . The method of claim 11 , wherein the simple starting material comprises a sugar.
13 . The method of claim 1 , further comprising:
contacting the codeinone product, or a derivative thereof, with one or more enzymes selected from the group consisting of: morphinone reductase, morphine dehydrogenase, codeinone reductase, N-demethylase, N-methyltransferase, and neopinone isomerase,
thereby producing at least one product selected from the group consisting of: a neopinone, 14-Hydroxycodeinone, oxycodone, oxymorphone, noroxymorphone, noroxymorphol, buprenorphine, naloxone, naltrexone, and nalbuphine.
14 . The method of claim 3 , further comprising:
contacting the hydrocodone product, or a derivative thereof, with one or more enzymes selected from the group consisting of: a morphine dehydrogenase, codeinone reductase, and codeine O-demethylase,
thereby producing at least one product selected from the group consisting of: hydromorphone and dihydrocodeine.
15 . The method of claim 4 , further comprising:
contacting the codeine product, or a derivative thereof, with one or more enzymes selected from the group consisting of: morphine dehydrogenase, codeinone reductase, codeine O-demethylase, and neopinone isomerase,
thereby producing at least one product selected from the group consisting of: a codeinone, neopinone, neopine, neomorphine, morphine, neomorphinone, and morphinone.
16 . The method of claim 1 , wherein the engineered non-plant cell is an engineered fungal cell or an engineered bacteria cell.
17 . The method of claim 16 , wherein the engineered non-plant cell is an engineered fungal cell.
18 . The method of claim 16 , wherein the engineered non-plant cell is an engineered bacteria cell.