LAUSR

Efficient conjugation between biomolecules and electrode materials is one of the main challenges in the field of biosensors. Cellobiose dehydrogenase (CDH) is a monomeric enzyme, which consists of two separate domains: one catalytic dehydrogenase domain (DHCDH ) carrying strongly bound flavin adenine dinucleotide (FAD) in the active site and a cytochrome domain (CYTCDH ) carrying a b-type heme connected by a flexible linker region. Herein, we report on the development of a lactose biosensor, based on direct electron transfer (DET) from CDH from Phanerochaete sordida (PsCDH) electrostatically attached onto polyethyleneimine-stabilized gold nanoparticles (PEI@AuNPs) used to cover a conventional polycrystalline solid gold disk electrode. PEI@AuNPs were synthesized in aqueous solution using PEI as reducing agent for AuIII and as stabilizer for the nanoparticles. The heterogeneous electron-transfer (ET) rate (ks ) for the redox reaction of immobilized PsCDH at the modified electrodes was calculated based on the Laviron theory and was found to be (39.6±2.5) s-1 . The proposed lactose biosensor exhibits good long term stability as well as high and reproducible sensitivity to lactose with a response time less than 5 s and a linear range from 1 to 100 μm.