Prasad Ankush, Kumar Aditya, Suzuki Makoto, Kikuchi Hiroyuki, Sugai Tomoya, Kobayashi Masaki, Pospíšil Pavel, Tada Mika, Kasai Shigenobu
Biomedical Engineering Research Center, Tohoku Institute of Technology Sendai, Japan.
Department of Biophysics, Faculty of Science, Centre of the Region Haná for Biotechnological and Agricultural Research, Palacký University Olomouc, Czech Republic.
Front Plant Sci. 2015 Oct 15;6:862. doi: 10.3389/fpls.2015.00862. eCollection 2015.
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.
已知过氧化氢（H₂O₂）通过酶促和非酶促途径在光系统II（PSII）中生成。人们已探索了用不同光谱技术检测H₂O₂，然而，其灵敏检测在光合研究中一直是个挑战。在最近，诸如Amplex Red（AR）等荧光探针已被使用，但已知其在H₂O₂的最低检测限方面缺乏特异性或存在局限性。我们采用了一种电化学生物传感器来实时监测亚细胞器水平上H₂O₂的生成。该电化学生物传感器由对电极和工作电极组成。对电极是一块铂板，而工作电极是一种基于介体的催化电流型生物传感器装置，它是通过用锇 - 辣根过氧化物酶涂覆碳电极而开发的，该酶可作为H₂O₂检测传感器。在当前研究中，已在光照条件下研究了PSII膜中H₂O₂的生成和动力学行为。使用催化电流型生物传感器装置对H₂O₂进行电化学检测被认为是一种用于检测光合细胞和包括PSII或类囊体膜在内的亚细胞结构中H₂O₂的有前景的技术。它还可以提供有关H₂O₂定性测定的精确信息，因此可广泛应用于光合研究。
