Asunto(s)
Oxidorreductasas de Alcohol/genética , Proteínas Bacterianas , Codeína/metabolismo , Hidrocodona , Hidromorfona , Morfina/metabolismo , Oxidorreductasas/genética , Biotransformación , Clonación Molecular , Escherichia coli , Plásmidos , Pseudomonas putida/enzimología , Pseudomonas putida/genéticaRESUMEN
The oxidation of morphine by washed-cell incubations of Pseudomonas putida M10 gave rise to a large number of transformation products including hydromorphone (dihydromorphinone), 14 beta-hydroxymorphine, 14 beta-hydroxymorphinone, and dihydromorphine. Similarly, in incubations with oxymorphone (14 beta-hydroxydihydromorphinone) as substrate, the major transformation product was identified as oxymorphol (14 beta-hydroxydihydromorphine). The identities of all these biological products were confirmed by mass spectrometry and 1H nuclear magnetic resonance spectroscopy. This is the first report describing structural evidence for the biological synthesis of 14 beta-hydroxymorphine and 14 beta-hydroxymorphinone. These products have applications as intermediates in the synthesis of semisynthetic opiate drugs.
Asunto(s)
Derivados de la Morfina/farmacocinética , Pseudomonas putida/metabolismo , Analgésicos Opioides/síntesis química , Analgésicos Opioides/química , Analgésicos Opioides/farmacocinética , Biotransformación , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Estructura Molecular , Derivados de la Morfina/síntesis química , Derivados de la Morfina/químicaRESUMEN
Semisynthetic derivatives of morphine and related alkaloids are in widespread clinical use. Due to the complexity of these molecules, however, chemical transformations are difficult to achieve in high yields. We recently identified the powerful analgesic hydromorphone as an intermediate in the metabolism of morphine by Pseudomonas putida M10. Here we describe the construction of recombinant strains of Escherichia coli that express morphine dehydrogenase and morphinone reductase. These strains are capable of efficiently transforming the naturally occurring alkaloids morphine and codeine to hydromorphone and the antitussive hydrocodone, respectively. Our results demonstrate the potential for recombinant DNA technology to provide biological routes for the synthesis of known and novel semisynthetic opiate drugs.
Asunto(s)
Oxidorreductasas de Alcohol/genética , Analgésicos Opioides/metabolismo , Proteínas Bacterianas , Ingeniería Genética , Oxidorreductasas/genética , Oxidorreductasas de Alcohol/biosíntesis , Codeína/metabolismo , Escherichia coli , Hidrocodona/metabolismo , Hidromorfona/metabolismo , Morfina/metabolismo , Oxidorreductasas/biosíntesis , Plásmidos/genética , Proteínas Recombinantes/biosíntesisAsunto(s)
Analgésicos/metabolismo , Antitusígenos/metabolismo , Proteínas Bacterianas , Ingeniería Genética , Oxidorreductasas de Alcohol/genética , Oxidorreductasas de Alcohol/metabolismo , Aldehído Reductasa/genética , Secuencia de Aminoácidos , Secuencia de Bases , Biotransformación , Clonación Molecular , ADN Bacteriano/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Genes Bacterianos , Hidrocodona/metabolismo , Hidromorfona/metabolismo , Datos de Secuencia Molecular , Morfina/química , Morfina/farmacocinética , Oxidorreductasas/metabolismo , Pseudomonas putida/genética , Pseudomonas putida/metabolismo , Homología de Secuencia de AminoácidoRESUMEN
The morphine alkaloid hydromorphone (dihydromorphinone) was identified as an intermediary metabolite in the degradation of morphine by Pseudomonas putida M10. A constitutive NADH-dependent morphinone reductase capable of catalyzing the reduction of the 7,8-unsaturated bond of morphinone and codeinone, yielding hydromorphone and hydrocodone, respectively, was shown to be present in cell extracts. The structures have been identified by 1H nuclear magnetic resonance and mass spectrometry. Morphinone reductase has been partially purified by anion-exchange and gel filtration chromatography. This enzyme has potential applications as a biocatalyst for the synthesis of the highly potent analgesic hydromorphone and the antitussive hydrocodone.
