Laccase immobilized on methylene blue modified mesoporous silica MCM-41/PVA

The mesoporous silica sieve MCM-41 containing methylene blue (MB) provides a suitable immobilization of biomolecule matrix due to its uniform pore structure, high surface areas, good biocompatibility and nice conductivity. Based on this, a facilely fabricated amperometric biosensor by entrapping laccase into the MB modified MCM-41/PVA composite film has been developed. Laccase from Trametes versicolor is assembled on a composite film of MCM-41 containing MB/PVA modified Au electrode and the electrode is characterized with respect to transmission electron microscopy (TEM) and scanning electron microscopic (SEM). Cyclic voltammetry (CV), response time, detection limit, linear range and activity of laccase. The laccase modified electrode remains good redox behavior in pH 4.95 acetate buffer solution, at room temperature in present of 0.1 mM catechol. The response time (t(90%)) of the modified electrode is less than 4 s for catechol. The detection limit is 0.331 mu M and the linear detect range is about from 4.0 mu M to 87.98 mu M for catechol with a correlation coefficient of 0.99913(S/N=3). The apparent Michaelis-Menten (K-M(app)) is estimated using the Lineweaver-Burk equation and the K-M(app) value is about 0.256 mM. This work demonstrated that the mesoporous silica MCM-41 containing MB provides a novel support for laccase immobilization and the construction of biosensors with a faster response and better bioactivity. Crown Copyright (C) 2009 Published by Elsevier B.V. All rights reserved.