title: A VOLTAMMETRIC FLAVIN MICROELECTRODE FOR USE IN BIOFILMS.
abstract: Biofilms used in bioelectrochemical systems are expected to transfer electrons using electron transfer mediators. One mediator type, flavins, which includes flavin mononucleotide, riboflavin, and flavin adenine dinucleotide, has been found to be endogenously produced by Shewanella oneidensis MR-1. However, the presence and concentration of flavins inside a S. oneidensis MR-1 biofilm have never been reported. The goal of this study was to develop a flavin microelectrode capable of measuring flavins inside a living biofilm and apply it to a biofilm which produces flavins. Because flavins are electrochemically active molecules, the flavin microelectrode was based on detection via square-wave voltammetry. The microelectrode consisted of a carbon working electrode with a 10-30 μm tip diameter, a built-in platinum counter electrode, and a Ag/AgCl reference electrode, all enclosed in a glass outer case. The microelectrode was calibrated between 0.1 μM and 10 μM flavins and showed a linear correlation between flavin concentration and peak currents located at -424 mV(Ag/AgCl) on a square-wave voltammogram. We also developed a model to explain the electrochemical mechanism of flavin detection, and to determine the effective surface area of the microelectrode, the standard reduction potential, and the transfer coefficient. We found that the effective surface area of the microelectrode was close to 100 times the projected surface area. The model predicted a standard reduction potential for RF/RFH2 of -419 mV(Ag/AgCl) at 20 °C and a transfer coefficient of 0.45. Lastly, we measured flavin concentration inside a S. oneidensis MR-1 biofilm grown on a glass surface using oxygen as the electron acceptor. The flavin concentration reached 0.7 μM, increasing near the bottom of the biofilm, where no oxygen was present. This shows the possibility that flavins are produced in the anaerobic zone to act as intermediate electron acceptors in the deeper parts of the biofilm, where there is no oxygen.
authors.affiliation: The Gene and Linda Voiland School of Chemical Engineering and Bioengineering, Washington State University, Pullman, WA, United States of America.
authors.forename: Hung Duc
authors.initials: HD
authors.lastname: Nguyen
authors.forename: Ryan
authors.initials: R
authors.lastname: Renslow
authors.forename: Jerome
authors.initials: J
authors.lastname: Babauta
authors.forename: Bulbul
authors.initials: B
authors.lastname: Ahmed
authors.forename: Haluk
authors.initials: H
authors.lastname: Beyenal