Co-immobilization of P450 BM3 and glucose dehydrogenase on different supports for application as a self-sufficient oxidative biocatalyst
Solé Ferré, Jordi (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Caminal i Saperas, Glòria (Institut de Química Avançada de Catalunya)
Schürmann, Martin (InnoSyn B.V. (The Netherlands))
Álvaro, Gregorio (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Guillén, Marina (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)

Date:	2019
Abstract:	BACKGROUND: The oxy-functionalization of non-activated carbon bonds by the bacterial cytochrome P450 BM3 from Bacillus megaterium, presents a promising field in biosynthesis and it has gainedmuchinterest in recent decades. Nevertheless, the need for the expensive cofactor NADPH, togetherwith lowoperational stability of the enzyme have made the implementation of this biocatalyst unfeasible in most cases for industry. RESULTS: P450 BM3 and glucose dehydrogenase (GDH), as a cofactor regeneration enzyme, were successfully co-immobilized obtaining a bi-functional self-sufficient oxidative biocatalyst. First, a broad screening of 13 different supports was carried out. Five selected agaroses with three different functionalities (epoxy, amine and aldehyde) were studied and their immobilization processes optimized. Finally, P450 BM3 and GDH were co-immobilized on those supports showing the best performance for P450 BM3 immobilization: epoxy-agarose (epoxy-agarose-UAB) presenting 83% and 20% retained activities respectively; AMINO-agarose presenting 28% and 25%, and Lentikats® with which both enzymes retained 100% of the initial activity. Furthermore, the re-utilization of the self-sufficient immobilized derivatives was tested in five repeated cycles. CONCLUSIONS: P450 BM3 andGDHhave been successfully immobilized on three supports and their re-usability has been tested in amodel reaction. It represents a step forward for future P450 BM3 industrial implementations.
Grants:	European Commission 635734 Agència de Gestió d'Ajuts Universitaris i de Recerca 2017/SGR-1462
Rights:	Tots els drets reservats.
Language:	Anglès
Document:	Article ; recerca ; Versió acceptada per publicar
Subject:	Immobilization ; Biocatalysis ; Enzymes ; Oxidation
Published in:	Journal of chemical technology and biotechnology, Vol. 94, issue 1 (Jan. 2019) , p. 244-255, p., ISSN 1097-4660

DOI: 10.1002/jctb.5770

Preprint 36 p, 870.4 KB

Postprint 38 p, 501.5 KB

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