Impresión molecular de enzimas con ligandos insolubles en agua para la biocatálisis no acuosa
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Published on: February 20, 2013
Joseph O Rich1, Vadim V Mozhaev, Jonathan S Dordick
1Biocatalysis Division, Albany Molecular Research Inc., 601 E. Kensington Road, Mt. Prospect, Illinois 60056, USA.
Journal of the American Chemical Society
|May 9, 2002
Resumen
Los investigadores mejoraron la actividad enzimática en disolventes orgánicos utilizando el imprinting molecular. Las estrategias superan los problemas de solubilidad de los ligandos, impulsando la catálisis enzimática para aplicaciones en la enzimología no acuosa.
Área de la Ciencia:
- Biocatálisis y ingeniería de enzimas.
- Química orgánica y efectos del disolvente.
- Biotecnología y aplicaciones industriales.
Sus antecedentes:
- La mejora de la actividad catalítica de las enzimas en disolventes orgánicos es crucial para la enzimología no acuosa.
- La impresión molecular es una estrategia exitosa para la sintonización del sitio activo de la enzima, pero está limitada por la solubilidad del impresor.
- La escasa solubilidad de muchos impresores importantes restringe la aplicación más amplia de esta técnica de modificación de enzimas.
Objetivo del estudio:
- Desarrollar estrategias para superar las limitaciones de solubilidad de la impresión molecular para la mejora de enzimas.
- Para ampliar la aplicabilidad de la impresión molecular más allá de los ligandos solubles en agua.
- Investigar técnicas combinadas de activación de enzimas para efectos sinérgicos.
Principales métodos:
- Estrategias para mejorar la solubilidad del impresor: conversión a formas solubles en agua o adición de cosolventes orgánicos (por ejemplo, alcohol tert-butilico, 1,4-dioxano).
- Aplicación de estas estrategias a la impresión molecular de la termolisina, la subtilisina y la lipasa TL.
- Evaluación de la actividad catalítica en reacciones de acilación (paclitaxel, 17beta-estradiol) y evaluación de los efectos combinados con la activación de la sal.
Principales resultados:
- Se produjeron con éxito enzimas impresas con una actividad catalítica significativamente mejorada (hasta 26 veces) en comparación con los controles no impresos.
- Demostró estrategias efectivas para superar los problemas de solubilidad del imprinter en el imprinting molecular.
- Se logró un efecto de activación aditiva sustancial (hasta 110 veces) mediante la combinación de la impresión molecular con la activación de la sal.
Conclusiones:
- Desarrolló estrategias versátiles para superar los desafíos de solubilidad del imprinter en el imprinting molecular para la mejora de enzimas.
- Aumentó significativamente la actividad catalítica de enzimas en disolventes orgánicos, ampliando la utilidad de la impresión molecular.
- Mostró el potencial sinérgico de combinar la impresión molecular con la activación de la sal para una activación enzimática sin precedentes.
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