Methods Of Improving Production Of Morphinan Alkaloids And Derivatives
AssigneeAntheia, Inc.
InventorsJames T. PAYNE, Amy M. KOZINA, Rodrigo A. ESTRADA, Kristy M. HAWKINS, Christina D. SMOLKE
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Abstract
Disclosed herein are methods that may be used for the synthesis of benzylisoquinoline alkaloids (“BIAs”) such as alkaloid morphinan. The methods disclosed can be used to produce thebaine, oripavine, codeine, morphine, oxycodone, hydrocodone, oxymorphone, hydromorphone, naltrexone, naloxone, hydroxycodeinone, neopinone, and/or buprenorphine.
Claims31
A method of producing a benzylisoquinoline alkaloid (BIA) product in an engineered host cell, the method comprising:
- (a) expressing a heterologous enzyme having 14-hydroxylase activity in the engineered host cell;
- (b) expressing a heterologous enzyme having cytochrome P450 reductase (CPR) activity in the engineered host cell,
- (c) contacting the heterologous enzyme with a BIA-precursor substrate that is a morphinan alkaloid with a free hydrogen at carbon C-14, wherein the heterologous enzyme hydroxylates the C-14 carbon on the BIA-precursor substrate;
- and
- (d) producing the BIA product within the host cell;
- wherein the engineered host cell produces more BIA product than a non-engineered host cell.
A host cell that produces a BIA product, the host cell comprising a first heterologous polynucleotide that encodes a heterologous enzyme having 14-hydroxylase activity and a second heterologous polynucleotide that encodes a cytochrome P450 reductase (CPR).
A vector comprising a first polynucleotide that encodes an enzyme having 14-hydroxylase activity and a second polynucleotide that encodes a cytochrome P450 reductase (CPR).
A host cell comprising a vector that comprises a first polynucleotide that encodes an enzyme having 14-hydroxylase activity and a second polynucleotide that encodes a cytochrome P450 reductase (CPR).
A host cell comprising a vector that comprises a first heterologous polynucleotide sequence that encodes a heterologous enzyme having formaldehyde dehydrogenase activity and a second heterologous polynucleotide designed to repress expression of a target gene, wherein the target gene is selected from DUG2 or DUG3.
Dependencies are inferred from the claim text.