Redesign of a Dioxygenase in Morphine Biosynthesis
Opium poppy (Papaver somniferum) produces medicinally important benzylisoquinoline alkaloids, including the analgesics codeine and morphine, in the morphinan pathway. We aligned three dioxygenases that were recently discovered in P. somniferum and subsequently identified the nonconserved regions. Tw...
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| Format: | Article |
| Language: | en_US |
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Elsevier
2014
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| Online Access: | http://hdl.handle.net/1721.1/91546 |
| _version_ | 1811069467017347072 |
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| author | Runguphan, Weerawat Glenn, Weslee S. O'Connor, Sarah Ellen |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Runguphan, Weerawat Glenn, Weslee S. O'Connor, Sarah Ellen |
| author_sort | Runguphan, Weerawat |
| collection | MIT |
| description | Opium poppy (Papaver somniferum) produces medicinally important benzylisoquinoline alkaloids, including the analgesics codeine and morphine, in the morphinan pathway. We aligned three dioxygenases that were recently discovered in P. somniferum and subsequently identified the nonconserved regions. Two of these enzymes, codeine O-demethylase (PsCODM) and thebaine O-demethylase (PsT6ODM), are known to facilitate regioselective O-demethylation in morphinan biosynthesis. We systematically swapped the residues that were nonconserved between the PsCODM and PsT6ODM sequences to generate 16 mutant PsCODM proteins that could be overexpressed in Escherichia coli. While wild-type PsCODM can demethylate both codeine and thebaine, one engineered PsCODM mutant selectively demethylates codeine. Use of this reengineered enzyme in the reconstitution of morphine biosynthesis could selectively disable a redundant pathway branch and therefore impact the yields of the downstream products codeine and morphine in subsequent metabolic engineering efforts. |
| first_indexed | 2024-09-23T08:10:54Z |
| format | Article |
| id | mit-1721.1/91546 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:10:54Z |
| publishDate | 2014 |
| publisher | Elsevier |
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| spelling | mit-1721.1/915462022-09-30T08:05:07Z Redesign of a Dioxygenase in Morphine Biosynthesis Runguphan, Weerawat Glenn, Weslee S. O'Connor, Sarah Ellen Massachusetts Institute of Technology. Department of Chemistry Runguphan, Weerawat Glenn, Weslee S. Opium poppy (Papaver somniferum) produces medicinally important benzylisoquinoline alkaloids, including the analgesics codeine and morphine, in the morphinan pathway. We aligned three dioxygenases that were recently discovered in P. somniferum and subsequently identified the nonconserved regions. Two of these enzymes, codeine O-demethylase (PsCODM) and thebaine O-demethylase (PsT6ODM), are known to facilitate regioselective O-demethylation in morphinan biosynthesis. We systematically swapped the residues that were nonconserved between the PsCODM and PsT6ODM sequences to generate 16 mutant PsCODM proteins that could be overexpressed in Escherichia coli. While wild-type PsCODM can demethylate both codeine and thebaine, one engineered PsCODM mutant selectively demethylates codeine. Use of this reengineered enzyme in the reconstitution of morphine biosynthesis could selectively disable a redundant pathway branch and therefore impact the yields of the downstream products codeine and morphine in subsequent metabolic engineering efforts. National Science Foundation (U.S.) (Predoctoral Fellowship) John Innes Centre University of East Anglia 2014-11-13T18:48:12Z 2014-11-13T18:48:12Z 2012-06 2012-04 Article http://purl.org/eprint/type/JournalArticle 10745521 http://hdl.handle.net/1721.1/91546 Runguphan, Weerawat, Weslee S. Glenn, and Sarah E. O’Connor. “Redesign of a Dioxygenase in Morphine Biosynthesis.” Chemistry & Biology 19, no. 6 (June 2012): 674–678. © 2012 Elsevier Ltd. en_US http://dx.doi.org/10.1016/j.chembiol.2012.04.017 Chemistry and Biology Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Elsevier Elsevier |
| spellingShingle | Runguphan, Weerawat Glenn, Weslee S. O'Connor, Sarah Ellen Redesign of a Dioxygenase in Morphine Biosynthesis |
| title | Redesign of a Dioxygenase in Morphine Biosynthesis |
| title_full | Redesign of a Dioxygenase in Morphine Biosynthesis |
| title_fullStr | Redesign of a Dioxygenase in Morphine Biosynthesis |
| title_full_unstemmed | Redesign of a Dioxygenase in Morphine Biosynthesis |
| title_short | Redesign of a Dioxygenase in Morphine Biosynthesis |
| title_sort | redesign of a dioxygenase in morphine biosynthesis |
| url | http://hdl.handle.net/1721.1/91546 |
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