RESUMEN
Herein we describe the ability of the permissive glycosyltransferase (GT) OleD Loki to convert a diverse set of >15 histone deacetylase (HDAC) inhibitors (HDACis) into their corresponding hydroxamate glycosyl esters. Representative glycosyl esters were subsequently evaluated in assays for cancer cell line cytotoxicity, chemical and enzymatic stability, and axolotl embryo tail regeneration. Computational substrate docking models were predictive of enzyme-catalyzed turnover and suggest certain HDACis may form unproductive, potentially inhibitory, complexes with GTs.
Asunto(s)
Proteínas Bacterianas/metabolismo , Glucosiltransferasas/metabolismo , Ácidos Hidroxámicos/metabolismo , Proteínas Bacterianas/antagonistas & inhibidores , Sitios de Unión , Biocatálisis , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Glucosiltransferasas/antagonistas & inhibidores , Glicosilación , Inhibidores de Histona Desacetilasas/química , Inhibidores de Histona Desacetilasas/metabolismo , Inhibidores de Histona Desacetilasas/farmacología , Humanos , Ácidos Hidroxámicos/química , Ácidos Hidroxámicos/farmacología , Simulación del Acoplamiento Molecular , Especificidad por SustratoRESUMEN
We describe the ability of an engineered glycosyltransferase (OleD Loki) to catalyze the N-glycosylation of tertiary-amine-containing drugs and trichostatin hydroxamate glycosyl ester formation. As such, this study highlights the first bacterial model catalyst for tertiary-amine N-glycosylation and further expands the substrate scope and synthetic potential of engineered OleDs. In addition, this work could open the door to the discovery of similar capabilities among other permissive bacterial glycosyltransferases.
Asunto(s)
Aminas/metabolismo , Glicosiltransferasas/química , Ácidos Hidroxámicos/química , Modelos Biológicos , Aminas/química , Catálisis , Dominio Catalítico/fisiología , Glicosilación , Glicosiltransferasas/genética , Estructura Molecular , Ingeniería de ProteínasRESUMEN
The assessment of glycosyl-scanning to expand the molecular and functional diversity of metabolites from the underground coal mine fire-associated Streptomyces sp. RM-14-6 is reported. Using the engineered glycosyltransferase OleD Loki and a 2-chloro-4-nitrophenylglycoside-based screen, six metabolites were identified as substrates of OleD Loki, from which 12 corresponding metabolite glycosides were produced and characterized. This study highlights the first application of the 2-chloro-4-nitrophenylglycoside-based screen toward an unbiased set of unique microbial natural products and the first reported application of the 2-chloro-4-nitrophenylglycoside-based transglycosylation reaction for the corresponding preparative synthesis of target glycosides. Bioactivity analysis (including antibacterial, antifungal, anticancer, and EtOH damage neuroprotection assays) revealed glycosylation to attenuate the neuroprotective potency of 4, while glycosylation of the structurally related inactive spoxazomicin C (3) remarkably invoked neuroprotective activity.
Asunto(s)
Antifúngicos/aislamiento & purificación , Antifúngicos/farmacología , Glicósidos/química , Fármacos Neuroprotectores/aislamiento & purificación , Fármacos Neuroprotectores/farmacología , Oligopéptidos/aislamiento & purificación , Oligopéptidos/farmacología , Oxazoles/aislamiento & purificación , Oxazoles/farmacología , Streptomyces/química , Antifúngicos/química , Glicosilación , Estructura Molecular , Fármacos Neuroprotectores/química , Oligopéptidos/química , Oxazoles/químicaRESUMEN
A set of 2-chloro-4-nitrophenyl glucosamino-/xylosaminosides were synthesized and assessed as potential substrates in the context of glycosyltransferase-catalyzed formation of the corresponding UDP/TDP-α-D-glucosamino-/xylosaminosugars and in single-vessel model transglycosylation reactions. This study highlights a robust platform for aminosugar nucleotide synthesis and reveals OleD Loki to be a proficient catalyst for U/TDP-aminosugar synthesis and utilization