Abstract
Multilayered films of cellulose nanoparticles (NFC’s) and modified multi-walled carbon nanotubes (MWCNT’s) were assembled by means of alternate electrostatic adsorption with positively charged poly(ethyleneimine) (PEI) onto cellulose support. The free carboxylic groups of NFC’s and MWCNT’s were coupled with ethylenediamine. Glucose oxidase and laccase were immobilized by means of Schiff base reaction between aldehyde groups of glutaraldehyde and the free amino sites of the proteins. The immobilized enzymes on the surface of nanoparticles have higher value for the specific activity compared to the enzymes immobilized directly on the cellulose surface indicating the stabilization of the proteins by the nanoparticles. The kinetics of the enzymes, catalyzed reactions and reusability of the enzymes were investigated and were showing better properties for enzymes cross-linked with glutaraldehyde. After 5 days the initial enzyme activity of glucose oxidase was around 85%, but the initial enzyme activity of laccase was 60%. The kinetic investigations of the immobilized enzymes showed no significant difference in Michaelis constant but the maximum reaction rate is decreased.
Highlights
Due to need of cheap, easy to use, biocompatible and biodegradable materials for biosensors and biofuel cell electrodes cellulose could offer some new alternatives as a material
Laccase from Trametes versicolor, glucose oxidase (GOx) from Aspergillus niger, glucose monohydrate, poly(ethyleneimine) solution with molecular weight 1200 -1300 Damköhler numbers (Da) (PEI), cellulose, glutaraldehyde, multi-walled carbon nanotubes, >90% carbon basis, DxL 110-170 nm x 5-9 nm, 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt 98% (ABTS), NaBr, NaClO, Triton X-100 were purchased from Sigma Aldrich. 2,2,6,6- Tetramethyl-1-piperidinyloxy, free radical 98% (TEMPO), 1-(3-Dimethylaminopropyl)-3ethylcarbodiimide hydrochloride, 98+% (EDC) were purchased from Alfa Aesar, Ethylenediamine 99%, extra pure was purchased from Acros Organics
Activity of the enzymes assembled onto Whatman 1 paper with PEI and glutaraldehyde
Summary
Due to need of cheap, easy to use, biocompatible and biodegradable materials for biosensors and biofuel cell electrodes cellulose could offer some new alternatives as a material. Cellulose is the most widely spread organic polymer found in nature, since it constitutes the main component of the membrane of plant cell. It is well-known that cellulose is a very important and fascinating biopolymer and an almost inexhaustible and renewable raw material [1]. On the other hand cellulose nanoparticles like nanofibrillated cellulose and cellulose nanocrystals can be applied as a support material with good chemical and mechanical properties for the preparation of carriers for enzyme immobilization
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