WO2016149821A8 - Methods of making morphinan alkaloids and enzymes therefore - Google Patents

Methods of making morphinan alkaloids and enzymes therefore Download PDF

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Publication number
WO2016149821A8
WO2016149821A8 PCT/CA2016/050334 CA2016050334W WO2016149821A8 WO 2016149821 A8 WO2016149821 A8 WO 2016149821A8 CA 2016050334 W CA2016050334 W CA 2016050334W WO 2016149821 A8 WO2016149821 A8 WO 2016149821A8
Authority
WO
WIPO (PCT)
Prior art keywords
metabolite
reticuline
enzymes
converts
making
Prior art date
Application number
PCT/CA2016/050334
Other languages
French (fr)
Other versions
WO2016149821A1 (en
Inventor
Elena Fossati
Lauren NARCROSS
Vincent Martin
Original Assignee
Valorbec Société en Commandite
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Valorbec Société en Commandite filed Critical Valorbec Société en Commandite
Publication of WO2016149821A1 publication Critical patent/WO2016149821A1/en
Publication of WO2016149821A8 publication Critical patent/WO2016149821A8/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
    • C12Y114/21—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14) with NADH or NADPH as one donor, and the other dehydrogenated (1.14.21)
    • C12Y114/21004—Salutaridine synthase (1.14.21.4)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
    • C12P17/188—Heterocyclic compound containing in the condensed system at least one hetero ring having nitrogen atoms and oxygen atoms as the only ring heteroatoms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
    • C12Y101/01248—Salutaridine reductase (NADPH) (1.1.1.248)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y106/00—Oxidoreductases acting on NADH or NADPH (1.6)
    • C12Y106/02—Oxidoreductases acting on NADH or NADPH (1.6) with a heme protein as acceptor (1.6.2)
    • C12Y106/02004—NADPH-hemoprotein reductase (1.6.2.4), i.e. NADP-cytochrome P450-reductase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y203/00—Acyltransferases (2.3)
    • C12Y203/01—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
    • C12Y203/0115—Salutaridinol 7-O-acetyltransferase (2.3.1.150)

Abstract

A method of preparing a morphinan alkaloid (MA) metabolite comprising: (a) culturing a host cell under conditions suitable for MA production including a first fermentation at a pH of between about 7.5 and about 10, said host cell comprising: (i) a first heterologous coding sequence encoding a first enzyme involved in a metabolite pathway that converts (R)-reticuline into the metabolite; (ii) a second heterologous coding sequence encoding a second enzyme involved in a metabolite pathway that converts (R)-reticuline into the metabolite; and/or (iii) a third heterologous coding sequence encoding a second enzyme involved in a metabolite pathway that converts (R)-reticuline into the metabolite; (b) adding (R)-reticuline to the cell culture; and (c) recovering the metabolite from the cell culture. Plasmids and host cells encoding the enzymes are also provided.
PCT/CA2016/050334 2015-03-23 2016-03-23 Methods of making morphinan alkaloids and enzymes therefore WO2016149821A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201562136912P 2015-03-23 2015-03-23
US62/136,912 2015-03-23

Publications (2)

Publication Number Publication Date
WO2016149821A1 WO2016149821A1 (en) 2016-09-29
WO2016149821A8 true WO2016149821A8 (en) 2017-04-06

Family

ID=56976979

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CA2016/050334 WO2016149821A1 (en) 2015-03-23 2016-03-23 Methods of making morphinan alkaloids and enzymes therefore

Country Status (1)

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WO (1) WO2016149821A1 (en)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2970999A2 (en) 2013-03-15 2016-01-20 The Board Of Trustees Of The University Of the Leland Stanford Junior University Benzylisoquinoline alkaloids (bia) producing microbes, and methods of making and using the same
CN112410379A (en) 2013-11-04 2021-02-26 小利兰·斯坦福大学托管委员会 Microorganisms producing benzylisoquinoline alkaloid (BIA) precursors and methods of making and using the same
CN107614688B (en) 2015-05-04 2023-02-28 小利兰·斯坦福大学托管委员会 Microorganisms producing benzylisoquinoline alkaloid (BIA) precursors and methods of making and using same
GB2555044B (en) 2015-05-08 2021-03-17 Univ Leland Stanford Junior Methods of producing epimerases and benzylisoquinoline alkaloids
JP2019536476A (en) 2016-10-18 2019-12-19 アンテイア インコーポレイテッド Method for producing nor-opioid and nal-opioid benzyl isoquinoline alkaloids
WO2019028390A1 (en) 2017-08-03 2019-02-07 Antheia, Inc. Engineered benzylisoquinoline alkaloid epimerases and methods of producing benzylisoquinoline alkaloids
WO2019051046A1 (en) * 2017-09-08 2019-03-14 Intrexon Corporation Microorganisms and methods in the fermentation of benzylisoquinoline alkaloids

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8975063B2 (en) * 2006-10-19 2015-03-10 California Institute Of Technology Compositions and methods for producing benzylisoquinoline alkaloids
WO2011058446A2 (en) * 2009-11-12 2011-05-19 Uti Limited Partnership Thebaine 6-o-demethylase and codeine o-demethylase from papaver somniferum
EP2970999A2 (en) * 2013-03-15 2016-01-20 The Board Of Trustees Of The University Of the Leland Stanford Junior University Benzylisoquinoline alkaloids (bia) producing microbes, and methods of making and using the same

Also Published As

Publication number Publication date
WO2016149821A1 (en) 2016-09-29

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