An improved amperometric triglyceride biosensor based on co-immobilization of nanoparticles of lipase, glycerol kinase and glycerol 3-phosphate oxidase onto pencil graphite electrode

Publication date: Available online 30 January 2017 Source:Enzyme and Microbial Technology Author(s): Vinay Narwal, C.S. Pundir Nanoparticles (NPs) of commercial lipase from Candida rugosa, of glycerol kinase (GK) from Cellulomonas species, of glycerol-3- phosphate oxidase (GPO) from Aerococcus viridans were prepared, characterized and co-immobilized onto a pencil graphite (PG) electrode. The morphological and electrochemical characterization of PG electrode was performed by scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) before and after co-immobilization of enzyme nanoparticles (ENPs). An improved amperometric triglyceride (TG) biosensor was fabricated using Lipase NPs/GKNPs/GPONPs/PG electrode as the working electrode, Ag/AgCl as the standard electrode and Pt wire as auxiliary electrode. The biosensor showed optimum response within 2.5s at a pH 7.0 and temperature of 35°C.The biosensor measured current due to electrons generated at 0.1V against Ag/AgCl, from H2O2, which is produced from triolein by co-immobilized ENPs. A linear relationship was obtained over between a wide triolein concentration range (0.1mM to 45mM) and current (mA) under optimal conditions.The LipaseNPs/GKNPs/GPONPs/PG electrode showed high sensitivity (1241 ±20mAcm−2 mM−1); a lower detection limit (0.1nM) and good correlation coeficient (R2 =0.99) with a standard enzymic colorimetric method. Analytical recovery of added triolein in se...
Source: Enzyme and Microbial Technology - Category: Biotechnology Source Type: research