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.

Production of Reactive Oxygen Species by Photosystem II as a Response to Light and Temperature Stress

“…In the spontaneous dismutation, the interaction of two O 2 •– is restricted due to repulsion of the negative charge on the molecule, whereas the interaction of the protonated form of superoxide, hydroperoxyl radical (HO 2 • ), either with O 2 •– or HO 2 • is feasible. Spontaneous dismutation has been recently monitored by real-time detection of H 2 O 2 in PSII membrane under high light using highly sensitive and selective osmium-horseradish modified electrode (Prasad et al, 2015). In the enzymatic dismutation, reduction and oxidation of O 2 •– is associated with the redox change of the redox active metal center which serves as a superoxide oxidase (SOO) and superoxide reductase (SOR), respectively.…”

Reactive Oxygen Species as a Response to Wounding: In Vivo Imaging in Arabidopsis thaliana

“…In photosynthetic organism, generation of reactive oxygen species (ROS) is a quite universal and fast defense mechanism, known to be associated with various stresses both in vivo and in vitro (Yadav and Pospíšil, 2012;Prasad et al, 2015;Prasad et al, 2016;Prasad et al, 2017a;Prasad et al, 2018;Kumar et al, 2019), in both local as well as systemic responses (Grant and Loake, 2000;Slesak et al, 2007). In tomato, it has been observed that hydrogen peroxide (H 2 O 2 ) was produced at the site within an hour of wounding and its level was enhanced even in distant part (upper unwounded leaves) in the following 4 to 6 h (Orozco-Cardenas and Ryan, 1999).…”

Plant Science View on Biohybrid Development

“…From a methodological point of view, the detection of ROS depends mainly on staining and imaging, which would not be easily applied in biohybrids (Steffens et al, 2013 ). Interestingly, constant development of electrochemical methods of H 2 O 2 detection in plant cells should be mentioned (Olvera-González et al, 2013 ; Prasad et al, 2015 ). Probably, they are more applicable in biohybrids, for they omit microscopic image acquisition and processing.…”