Laccase immobilization on bacterial nanocellulose membranes: antimicrobial, kinetic and stability properties

This work studied the physical immobilization of a commercial laccase on bacterial nanocellulose (BNC) aiming to identify the laccase antibacterial properties suitable for wound dressings. Physico-chemical analysis demonstrates that the BNC structure is manly formed by pure crystalline I cellulose. The pH optimum and activation energy of free laccase depends on the substrate employed corresponding to pH 6, 7, 3 and 57, 22, 48 kJ mol1 for 2,6-dimethylphenol (DMP), catechol and 2,2 -azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), respectively. The Michaelis-Menten constant (Km) value for the immobilized laccase (0.77 mM) was found to be almost double of that of the free enzyme (0.42 mM). However, the specific activities of immobilized and free laccase are similar suggesting that the cage-like structure of BNC allows entrapped laccase to maintain some flexibility and favour substrate accessibility. The results clearly show the antimicrobial effect of laccase in Gram-positive (92%) and Gram-negative (26%) bacteria and cytotoxicity acceptable for wound dressing applications.

Andrea Zille (C2011-UMINHO-2C2T-01) acknowledges FCT funding from Programa Compromisso para a Ciência 2008, Portugal, FEDER funding on the Programa Operacional Factores de Competitividade - COMPETE and by national funds through FCT – Foundation for Science and Technology within the scope of the project POCI-01-0145-FEDER-007136 and UID/CTM/00264. Jorge Padrão and João P. Silva acknowledge Portuguese Foundation for Science and Technology (FCT) grants SFRH/BD/64901/2009 and SFRH/BPD/64958/2009 respectively. The authors Jorge Padrão, JoãoP. Silva and Fernando Dourado would like to thank the FCT StrategicProject PEst-OE/EQB/LA0023/2013, also to the Project “BioHealth −Biotechnology and Bioengineering approaches to improve healthquality”, Ref. NORTE-07-0124-FEDER-000027, co-funded by thePrograma Operacional Regional do Norte (ON.2-O Novo Norte),QREN, FEDER, and finnally to RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462).