A novel mıcrobıal bıosensor system based on C. tropicalis yeast cells for selectıve determınatıon of L-Ascorbıc acid

Abstract

In this study, a novel microbial biosensor was developed for the selective determination of L-Ascorbic acid. In the construction of the microbial biosensor, lyophilized Candida tropicalis yeast cells were immobilized with o-aminophenol by forming a film layer on a platinum electrode surface using electropolymerization. L-Ascorbic acid was quantified on the basis of both amperometric and differential pulse voltammetry (DPV) methods using the biosensor. The measurements were made at +0.24 V (vs Ag/AgCl) for amperometric studies and between 0.0 V and +0.7 V for DPV studies based on the oxidation of L-Ascorbic acid to dehydro-L-Ascorbic acid by ascorbate oxidase which takes place within the catabolic metabolic pathway of C. tropicalis yeast cells. According to the results obtained from the two methods, the response of the biosensor depends linearly on L-Ascorbic acid concentration between 100 and 1500 μM. The detection limit was 62 μM and 59 μM for amperometric and DPV measurements, respectively. The response time of the microbial biosensor was 14 s and 5 s for DPV and amperometric measurements, respectively. In the optimization studies of the biosensor, some parameters such as the optimum amount of the microorganism, o-aminophenol concentration, pH and temperature were determined. For the characterization of the biosensor, reproducibility, storage stability and the effect of interferences were determined.