Microbial degradation of the morphine alkaloids: identification of morphinone as an intermediate in the metabolism of morphine by Pseudomonas putida M10

Bruce, Neil C.; Wilmot, C J; Jordan, K N; Trebilcock, A E; Stephens, L D Gray; Lowe, Christopher R.

doi:10.1007/bf00245229

Cited by 53 publications

(43 citation statements)

References 16 publications

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“…Pseudomonas putida M10, was originally isolated by selective enrichment of industrial waste liquors [3] and was grown in the same manner as described in [7]. Media contained 20 mM glucose or 50 mM acetate as a carbon and energy source.…”

Section: Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

“…Cell-free extracts of P. putida M10 were prepared by sonic disruption followed by centrifugation [3]. Membrane preparations were prepared by centrifuging cell-free extract at 125 000Ug for 1 h in a Beckman Optima TLX ultracentrifuge using a TLA 45 rotor at 4³C.…”

Section: Preparation Of Cell-free Extract and Membrane Preparationsmentioning

confidence: 99%

“…Bruce et al [2] have reported the metabolism of morphine alkaloids by Pseudomonas putida M10 and the use of speci¢c enzymes isolated from this organism for the production of semi-synthetic opiate drugs. This organism was isolated from opiate waste liquors by its ability to utilise morphine and codeine as carbon sources for growth, albeit very slowly [3]. The ¢rst enzyme in the degradative pathway, morphine dehydrogenase, catalyses the NADP -dependent oxidation of morphine to morphinone and codeine to codeinone.…”

Section: Introductionmentioning

confidence: 99%

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Transformations of codeine to important semisynthetic opiate derivatives by Pseudomonas putida m10

Lister¹

1999

FEMS Microbiology Letters

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A biotransformation mixture which contained codeine and washed cells of Pseudomonas putida M10 gave rise to a number of transformation products that are of clinical importance which included hydrocodone, dihydrocodeine and 14L-hydroxycodeine. Incubations with the same organism and codeinone gave rise to 14L-hydroxycodeinone and 14L-hydroxycodeine. Cell-free extracts and membrane fractions of P. putida M10 were shown to catalyse the 14L-hydroxylation of codeinone. In addition, the potent analgesic oxycodone was shown to be produced from 14L-hydroxycodeinone. ß

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Section: Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

Section: Preparation Of Cell-free Extract and Membrane Preparationsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Transformations of codeine to important semisynthetic opiate derivatives by Pseudomonas putida m10

Lister¹

1999

FEMS Microbiology Letters

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“…The whole-cell recombination process is mediated by two constitutively expressed Pseudomonas enzymes, an NADP ϩ -dependent morphine dehydrogenase (MDH) and an NADH-dependent morphinone reductase (MR) (5, 6,8,12) (Fig. 1).…”

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confidence: 99%

Cofactor Regeneration by a Soluble Pyridine Nucleotide Transhydrogenase for Biological Production of Hydromorphone

Boonstra

Rathbone

French

et al. 2000

Appl Environ Microbiol

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We have applied the soluble pyridine nucleotide transhydrogenase of Pseudomonas fluorescens to a cell-free system for the regeneration of the nicotinamide cofactors NAD and NADP in the biological production of the important semisynthetic opiate drug hydromorphone. The original recombinant whole-cell system suffered from cofactor depletion resulting from the action of an NADP ؉ -dependent morphine dehydrogenase and an NADHdependent morphinone reductase. By applying a soluble pyridine nucleotide transhydrogenase, which can transfer reducing equivalents between NAD and NADP, we demonstrate with a cell-free system that efficient cofactor cycling in the presence of catalytic amounts of cofactors occurs, resulting in high yields of hydromorphone. The ratio of morphine dehydrogenase, morphinone reductase, and soluble pyridine nucleotide transhydrogenase is critical for diminishing the production of the unwanted by-product dihydromorphine and for optimum hydromorphone yields. Application of the soluble pyridine nucleotide transhydrogenase to the whole-cell system resulted in an improved biocatalyst with an extended lifetime. These results demonstrate the usefulness of the soluble pyridine nucleotide transhydrogenase and its wider application as a tool in metabolic engineering and biocatalysis.The pyridine nucleotide cofactors NAD and NADP are essential components of the cell, where they act as electron carriers in reduction and oxidation reactions. A large percentage of enzymes are dependent on these coenzymes for their activities (SwissProt database, http://www.expasy.ch/sprot/), and the cofactors are known to be involved in a vast amount of reactions (EcoCyc database). Many of the NAD(P)-dependent oxidoreductases catalyze reactions of commercial interest and have many applications, for instance, in the production of chiral compounds, amino acids, steroids, and other therapeutics for the pharmaceutical industry, in the modification or synthesis of polymers, in the oxidative remediation of pollutants, in the oxyfunctionalization of hydrocarbons, and in the construction of biosensors (14,18). The high cost of the pyridine nucleotide cofactors which need to be provided in stoichiometric quantities in enzyme reactions is an important commercial issue. Cofactor regeneration is, therefore, an important consideration when processes involving NAD(P)-dependent oxidoreductases are to be applied in a commercial setting.In cell-free systems the cofactors must be supplied, albeit at a lower-than-stoichiometric concentration (catalytic amounts), when cofactor regeneration is achieved. Alternatively, processes dependent on these cofactors can be carried out in whole cells which are known to have some reserves of the cofactor, but also here cofactor depletion can be a problem. Mostly NAD is regenerated using formate dehydrogenase, while NADP is recycled using glucose dehydrogenase. Recently, Galkin et al. described the synthesis of optically active amino acids from 2-keto acids using recombinant Escherichia coli coexpressing an amino ...

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“…These transformations have mainly comprised either N-dealkylation, reduction of the C-7-C-8 vinylic group, oxidation or reduction of the C-6 oxygroup, or hydroxylation of the C-14 position of the morphinan skeleton (10,11,14,15,21,22). Several of the morphine alkaloid-transforming enzymes have been purified and characterized (2,3,7,16). Furthermore, the genes encoding a number of these enzymes have been cloned and expressed in Escherichia coli, which has facilitated the engineering of recombinant cells to produce biologically semisynthetic opiate drugs (1,8,9,20).…”

mentioning

confidence: 99%

Transformation of 2,2′-Bimorphine to the Novel Compounds 10-α- S -Monohydroxy-2,2′-Bimorphine and 10,10′-α,α′- S , S ′-Dihydroxy-2,2′-Bimorphine by Cylindrocarpon didymum

Stabler

Holt

Bruce

2001

Appl Environ Microbiol

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Microbial degradation of the morphine alkaloids: identification of morphinone as an intermediate in the metabolism of morphine by Pseudomonas putida M10

Cited by 53 publications

References 16 publications

Transformations of codeine to important semisynthetic opiate derivatives by Pseudomonas putida m10

Transformations of codeine to important semisynthetic opiate derivatives by Pseudomonas putida m10

Cofactor Regeneration by a Soluble Pyridine Nucleotide Transhydrogenase for Biological Production of Hydromorphone

Transformation of 2,2′-Bimorphine to the Novel Compounds 10-α- S -Monohydroxy-2,2′-Bimorphine and 10,10′-α,α′- S , S ′-Dihydroxy-2,2′-Bimorphine by Cylindrocarpon didymum

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