A high sensitivity amperometric biosensor using laccase as biorecognition element.
An amperometric flow biosensor, using laccase from Rigidoporus lignosus as bioelement was developed. The laccase was kinetically characterized towards various phenolics both in solution and immobilized to a hydrophilic matrix by carbodiimide chemistry. A bioreactor connected to an amperometric flow cell by a FIA system was filled with the immobilized enzyme and the operational conditions of this biosensor were optimized as regards pH. Under the adopted experimental conditions, the immobilized enzyme oxidizes all the substrate molecules avoiding the need of cumbersome calibration procedures. The biosensor sensitivity, which was found to be 100 nA/microM for some of the tested substrates, resulted to be constant for more than 100 working days. This biosensor permits the detection of phenolics in aqueous solutions at concentrations in the nanomolar range and was successfully used to detect phenolics in wastewaters from olive oil mill without sample preparation.
Vianello
Fabio
F
Department of Biological Chemistry, University of Padua, Viale G. Colombo 3, 35121 Padova, Italy.
Ragusa
Santa
S
Cambria
Maria Teresa
MT
Rigo
Adelio
A
D015374:Biosensing Techniques / Q000295:instrumentation*; D004553:Electric Conductivity; D004563:Electrochemistry; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007220:Industrial Waste / Q000032:analysis; D042845:Laccase; D019800:Phenol / Q000032:analysis*; D013149:Spiroplasma / Q000201:enzymology; D014874:Water Pollutants, Chemical / Q000032:analysis
D007220:Industrial Waste; D014874:Water Pollutants, Chemical; D019800:Phenol; D042845:Laccase