Computer-aided laccase engineering: toward biological oxidation of arylamines

Oxidation of arylamines, such as aniline, is of high industrial interest and laccases have been proposed as biocata-lysts to replace harsh chemical oxidants. However, the reaction is hampered by the redox potential of the substrate at acid pH and enzyme engineering is required to improve the oxidati...

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Detalles Bibliográficos
Autores: Santiago, Gerard, de Salas, Felipe, Lucas, Fatima, Monza, Emanuele, Acebes, Sandra, Martinez, Ángel T., Camarero, Susana
Tipo de recurso: artículo
Fecha de publicación:2016
País:España
Institución:Universitat Politècnica de Catalunya (UPC)
Repositorio:UPCommons. Portal del coneixement obert de la UPC
Idioma:inglés
OAI Identifier:oai:upcommons.upc.edu:2117/88711
Acceso en línea:https://hdl.handle.net/2117/88711
https://dx.doi.org/10.1021/acscatal.6b01460
Access Level:acceso abierto
Palabra clave:Protein engineering
Oxidation--Measurement
Proteins
Computer-aided design
Laccase
Aniline
PELE
QM/MM
PANI
Arylamines
Oxidació
Enginyeria de proteïnes
Àrees temàtiques de la UPC::Enginyeria electrònica
Descripción
Sumario:Oxidation of arylamines, such as aniline, is of high industrial interest and laccases have been proposed as biocata-lysts to replace harsh chemical oxidants. However, the reaction is hampered by the redox potential of the substrate at acid pH and enzyme engineering is required to improve the oxidation. In this work, instead of trying to improve the redox potential of the en-zyme, we aim towards the (transient) substrate’s one and propose this as a more reliable strategy. We have successfully combined a computational approach with experimental validation to rationally design an improved biocatalyst. The in silico protocol combines classical and quantum mechanics to deliver atomic and electronic level detail on the two main processes involved: substrate binding and electron transfer. After mutant expression and comparison to the parent type, kinetic results show that the protocol accurately predicts aniline’s improved oxidation (2-fold kcat increase) in the engineered variant for biocatalyzed polyaniline production.