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Ingeniería de una [FeFe]-hidrogenasa: ¿Los grupos accesorios influyen en la resistencia y el sesgo catalítico?

Giorgio Caserta¹, Cecilia Papini¹, Agnieszka Adamska-Venkatesh²

¹Laboratoire de Chimie des Processus Biologiques , Collège de France, Université Pierre et Marie Curie, CNRS UMR 8229, PSL Research University , 11 place Marcelin Berthelot , 75005 Paris , France.

Journal of the American Chemical Society

|March 30, 2018

Resumen

Las hidrogenasas [FeFe] modificadas (HydAs) muestran una mayor resistencia al oxígeno y actividad de reducción de protones. Una versión truncada (MeH-HydA) conserva las características clave, ofreciendo un biocatalizador más simple y robusto para la producción de hidrógeno.

Área de la Ciencia:

Biocatálisis
Ingeniería de enzimas
Producción de biohidrógeno

Sus antecedentes:

Las [FeFe]-hidrogenasas (HydAs) son cruciales para la producción de H2, pero están limitadas por la sensibilidad y la complejidad del oxígeno.
Megasphaera elsdenii HydA (MeHydA) muestra una notable resistencia al oxígeno.
Los grupos accesorios de hierro y azufre en HydAs contribuyen a su complejidad.

Objetivo del estudio:

Para diseñar una hidrogenasa [FeFe] simplificada y resistente al oxígeno.
Investigar las propiedades catalíticas de un MeHydA truncado (MeH-HydA) que carece de grupos accesorios.
Para mejorar la actividad de la hidrogenasa y el sesgo de reducción de protones.

Principales métodos:

Truncamiento de proteínas para crear MeH-HydA.
Ensamblaje in vitro del grupo H activo.
Caracterización espectroscópica y bioquímica.
Ensayos de actividad enzimática para la hidrogenasa y la reducción de protones.

Principales resultados:

Ensamblaje funcional del sitio activo en MeH-HydA in vitro.
MeH-HydA retiene la resistencia al oxígeno de MeHydA.
El truncamiento mejora el sesgo de reducción de protones al alterar los pasos catalíticos que limitan la velocidad.
MeH-HydA exhibe una excelente actividad de la hidrogenasa.

Conclusiones:

Los HydAs diseñados pueden combinar la resistencia al oxígeno con una estructura simplificada.
MeH-HydA ofrece un biocatalizador robusto y eficiente para la producción de hidrógeno.
Este trabajo allana el camino para enzimas de hidrogenasa más simples y resistentes.

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