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

Abstract

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 microM, 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.

Biomimetic biosensor based on lipidic layers containing tyrosinase and lutetium bisphthalocyanine for the detection of antioxidants

“…tivity, sensitivity and potential for miniaturization (di Fusco et al, 2010;Gutés et al, 2005). A variety of foods and beverages such as beer (El Kaoutit et al, 2007), tea (Ghindilis et al, 1992), vegetable extracts (Franzoi et al, 2009), olive oil (Busch et al, 2006) and wine (Sanz et al, 2005) have been analysed using amperometric biosensors.…”

Novel Coulometric Approach to Evaluation of Total Free Polyphenols in Tea and Coffee Beverages in Presence of Milk Proteins

“…Biosensor containing enzyme tyrosinase immobilized by the crosslinking method has been used for detection of polyphenols in tea (Abhijith et al 2007). Biosensor based on the biimmobilization of laccase and tyrosinase phenoloxidase enzymes and the Sonogel-Carbon as electrochemical transducer has been developed for estimation of the total polyphenol index in beer samples (ElKaoutit et al 2007). …”

Bioelectronic Tongues Employing Electrochemical Biosensors

“…14, showing how the cross response may be easily achieved; as it can be seen in there, laccase may show increased response for caffeic acid or catechol, both being o-diphenols, or phenol will show large signal increment by coupling the two enzymes. This idea was later retaken by El-Kaoutit et al to evaluate main polyphenols present in beer and to estimate total polyphenol index in beer samples [51]. Final results obtained are summarized in Fig.…”

“…15 The use of the sensor array as key concept in the development of an electronic tongue analysis system. Adapted from the data in [51], with permission from American Chemical Society Fig. 14 Cross-sensitivity to selected phenolic compounds shown by phenol-degrading enzymes used in biosensors alone or associated in pair.…”