Polynorepinephrine and polydopamine-bacterial laccase coatings for phenolic amperometric biosensors

The successful fabrication of biosensors is greatly limited by the immobilization of their bioreceptor, thus we propose a facile and reproducible two-step method to modify graphite electrodes with a bacterial laccase, relying on a fast and controllable potentiostatic process to coat graphite surfaces with biomolecule-compatible thin films of polynorepinephrine (ePNE) and polydopamine (ePDA). Both polymers, synthesized with a similar thickness, were functionalized with bacterial laccase, displaying distinct electrochemical transducing behaviours at pH 5.0 and 7.0. ePNE layer enables adequate electron transfer of anionic and cationic species in acidic and neutral media, whereas transduction across ePDA strongly depends on pH and redox probe charge. ePNE stands out by improving the amperometric responses of the biosensing interface towards a phenolic acid (gallic acid) and a flavonoid (catechin), in respect to ePDA. The optimal graphite/ePNE/laccase interface outperforms biosensing interfaces based on fungal laccases at neutral pH, displaying detection sensitivities of 104 and 14.4 µA cm⁻² mM⁻¹ for gallic acid and catechin, respectively. The fine synthetic control of the ePNE bio-inspired transduction layer and the use of an alkaliphilic bacterial laccase enabled the construction of an amperometric biosensing interface with extended pH range of polyphenols detection present in food products and agro-industrial waste.