Dual Laccase–Tyrosinase Based Sonogel–Carbon Biosensor for Monitoring Polyphenols in Beers

• Published in: Journal of agricultural and food chemistry Vol. 55; no. 20; pp. 8011 - 8018
• Main Authors: ElKaoutit, Mohammed, Naranjo-Rodriguez, Ignacio, Temsamani, Khalid Riffi, de la Vega, Manuel Domínguez, de Cisneros, Jose Luis Hidalgo-Hidalgo
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 03.10.2007
• Subjects: amperometric biosensor; Analytical Methods; Beer - analysis; Beers; bioelectrochemical index; Biological and medical sciences; Biosensing Techniques - instrumentation; Biosensors; Biotechnology; Chemical Phenomena; Chemistry, Physical; dual laccase-tyrosinase based sonogel-carbon biosensor; Electrodes; enzyme inactivation; Enzyme Stability; Enzymes, Immobilized; Fermented food industries; Flavonoids - analysis; food analysis; food composition; Food industries; Fundamental and applied biological sciences. Psychology; immobilized enzymes; Kinetics; Laccase; Methods. Procedures. Technologies; monitoring; Monophenol Monooxygenase; new products; Phenols - analysis; Polyphenols; Reproducibility of Results; signal bioamplification; Sonogel–Carbon electrode; Trametes versicolor; Various methods and equipments; polyphenols; amperometric biosensor; bioelectrochemical index; signal bioamplification; Sonogel–Carbon electrode; beers; Bass; Edible fish; Enzyme; Index; Sonogel-Carbon electrode; Carbon; Beer; Electrodes; Alcoholic beverage; Polyphenol; Biosensor; Oxidoreductases; Monitoring; Laccase
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf0711136

A biosensor based on the bi-immobilization of laccase and tyrosinase phenoloxidase enzymes has been successfully developed. This biosensor employs as the electrochemical transducer the Sonogel–Carbon, a novel type of electrode developed by our group. The immobilization step was accomplished by doping the electrode surface with a mixture of the enzymes, glutaricdialdehyde, and Nafion-ion exchanger, as protective additive. The response of this biosensor, denoted the dual Trametes versicolor laccase (La) and Mushroom tyrosinase (Ty) based Sonogel–Carbon, was optimized directly in beer real samples and its analytical performance with respect to five individual polyphenols was evaluated. The Lac–Ty/Sonogel–Carbon electrode responds to nanomolar concentrations of flavan-3-ols, hydroxycinnamic acids, and hydroxybenzoic acids. The limit of detection, sensitivity, and linear range for caffeic acid, taken as an example, were 26 nM, 167.53 nA M−1, and 0.01–2 µM, respectively. In addition, the stability and reproducibility of the biosensor were also evaluated in beer samples. The Lac–Ty/sonogel–carbon electrode was verified as very stable in this matrix, maintaining 80% of its stable response for at least three weeks, with a RSD of 3.6% (n = 10). The biosensor was applied to estimate the total polyphenol index in ten beer samples, and a correlation of 0.99 was obtained when the results were compared with those obtained using the Folin–Ciocalteau reagent.