The characterization of ordered thin films built of immobilized phenoloxidases

Novel immobilization method based on thin Langmuir–Blodgett film as a matrix has been developed for characterization of sensing protein layers effect. Often biomolecules can be incorporated and immobilized into Langmuir–Blodgett film by using covalent immobilization of proteins with glutaraldehyde. Laccase from Cerrena unicolor and tyrosinase from Agaricus bisporus were incorporated into LB film deposited on glass substrate which can be used for phenolic compounds detection. The function of enzyme immobilization was carried by glutaraldehyde added onto the film of stearic acid and phenoloxidases (laccase, tyrosinase). The sensor sensitization was achieved by an amphiphilic N-alkyl-bis(thiophene)diphenylamine admixed into the film. The interlaced diphenylamine derivative was expected to facilitate the electron transfer thereby enhancing the sensor sensitivity. This immobilization strategy showed high efficiency. These results suggest that it is very promising for exploring interaction between proteins and high throughput detecting the phenol derivatives in wastewater.