Abstract

The capacity to generate a constant signal response from an enzyme on an electrode surface has been a fasci-nating topic of research from the past three decades. To nourish the enzymatic activity during electrochemical reactions, the immobilization of dual enzymes on the electrode surface could prevent the enzymatic loss without denaturation and thus long-term stability can be achieved. For effective immobilization of dual enzymes, mes-oporous materials are the ideal choice because of its numerous advantages such as 1. The presence of porous structure facilitates high loading of enzymes 2. The formation of protective environment can withstand the enzymatic activity even at acidic or basic pH values and even at elevated temperatures. Herein, we develop bienzymatic immobilization of horseradish peroxidase (HRP) and cholesterol oxidase (ChOx) on mesoporous V2O5-TiO2 based binary nanocomposite for effective sensing of hydrogen peroxide (H2O2) in presence of redox mediator hydroquinone (HQ). The utilization of redox mediator in second-generation biosensing of H2O2 can eliminate the interference species and reduces the operating potential with higher current density for electro-chemical reduction reaction. Using this mediator transfer process approach at HRP/ChOx/V2O5-TiO2 modified GC, the H2O2 can be determined at operating potential (-0.2 V) with good linear range (0.05-3.5 mM) higher sensitivity (1040 mu A mu M-1 cm-2) and lower detection limit of about 20 mu M can be attained, which is due to higher mediation of electrons were transferred to the enzyme cofactors. These interesting characteristics could be due to mesoporous structure of V2O5-TiO2 can induce large immobilization and facilitate higher interaction with en-zymes for wide range of biosensing applications.