Development of biosensor for phenol detection using agarose–guar gum based laccases extracted from Pleurotus ostreatus

Laccases from Pleurotus ostreatus was extracted from the Shaken flask cultures of Pleurotus ostreatus grown at 25°C with continuous agitation (110 rpm.) in baffled 1000 mL Erlenmeyer flasks containing 200 mL medium. The basal GYP medium used for cultures contained 20 g glucose l^?1, 5 g yeast extract l^?1, 5 g peptone from casein l^?1, and 1 g MgSO₄.7H₂O l^?1. The pH was adjusted to 5.0 with H₃PO₄ before sterilization. The kinetics of oxidation reactions catalyzed by laccases was studied using 2,2′‐azino‐bis (3‐ethylbenzthiazoline‐6‐sulphonic acid). The laccases showed lower specific activity and higher activity in nonpolar organic solvents. A biosensor using laccases was constructed for the determination of phenol. The enzyme was extracted from Pleurotus ostreatus and entrapped in agarose–guar gum composite biopolymer matrix. Phenol was determined by direct reduction of biocatalytically liberated quinone species at ?0.1 V versus Ag/AgCl (3M KCl). The response was found to be linear and concentration dependent.It has a shelf life of more than 2 months when stored at 4°C. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010