title: Detection of hydrogen peroxide in Photosystem II (PSII) using catalytic amperometric biosensor.
abstract: Hydrogen peroxide (H2O2) is known to be generated in Photosystem II (PSII) via enzymatic and non-enzymatic pathways. Detection of H2O2 by different spectroscopic techniques has been explored, however its sensitive detection has always been a challenge in photosynthetic research. During the recent past, fluorescence probes such as Amplex Red (AR) has been used but is known to either lack specificity or limitation with respect to the minimum detection limit of H2O2. We have employed an electrochemical biosensor for real time monitoring of H2O2 generation at the level of sub-cellular organelles. The electrochemical biosensor comprises of counter electrode and working electrodes. The counter electrode is a platinum plate, while the working electrode is a mediator based catalytic amperometric biosensor device developed by the coating of a carbon electrode with osmium-horseradish peroxidase which acts as H2O2 detection sensor. In the current study, generation and kinetic behavior of H2O2 in PSII membranes have been studied under light illumination. Electrochemical detection of H2O2 using the catalytic amperometric biosensor device is claimed to serve as a promising technique for detection of H2O2 in photosynthetic cells and subcellular structures including PSII or thylakoid membranes. It can also provide a precise information on qualitative determination of H2O2 and thus can be widely used in photosynthetic research.
authors.affiliation: Biomedical Engineering Research Center, Tohoku Institute of Technology Sendai, Japan.
authors.forename: Ankush
authors.initials: A
authors.lastname: Prasad
authors.affiliation: Department of Biophysics, Faculty of Science, Centre of the Region Haná for Biotechnological and Agricultural Research, Palacký University Olomouc, Czech Republic.
authors.forename: Aditya
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Graduate Department of Environmental Information Engineering, Tohoku Institute of Technology Sendai, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Suzuki
authors.affiliation: Graduate Department of Environmental Information Engineering, Tohoku Institute of Technology Sendai, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kikuchi
authors.affiliation: Graduate Department of Environmental Information Engineering, Tohoku Institute of Technology Sendai, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Sugai
authors.affiliation: Biomedical Engineering Research Center, Tohoku Institute of Technology Sendai, Japan ; Graduate Department of Electronics, Tohoku Institute of Technology Sendai, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Kobayashi
authors.affiliation: Department of Biophysics, Faculty of Science, Centre of the Region Haná for Biotechnological and Agricultural Research, Palacký University Olomouc, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Pospíšil
authors.affiliation: Biomedical Engineering Research Center, Tohoku Institute of Technology Sendai, Japan ; Center for General Education, Tohoku Institute of Technology Sendai, Japan.
authors.forename: Mika
authors.initials: M
authors.lastname: Tada
authors.affiliation: Biomedical Engineering Research Center, Tohoku Institute of Technology Sendai, Japan ; Graduate Department of Environmental Information Engineering, Tohoku Institute of Technology Sendai, Japan.
authors.forename: Shigenobu
authors.initials: S
authors.lastname: Kasai
keywords: EPR-spin trapping; amperometric biosensor; hydrogen peroxide; photosystem II; reactive oxygen species; superoxide anion radical