Helmholtz Centre for Infection Research, Working Group BiSA, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Biosens Bioelectron. 2013 Nov 15;49:192-8. doi: 10.1016/j.bios.2013.05.015. Epub 2013 May 15.
Despite advances made in the field, rapid detection methods for the human pathogen Candida albicans are still missing. In this regard, bio-electrochemical systems including electrochemical sensors and biosensors satisfy the increasing demand for rapid, reliable, and direct microbial analyses. In this study, the bioelectrochemical characteristics of C. albicans were investigated for use in an analytical system that determines the viability of the organisms. The electrochemical responses of viable and non-viable cells of C. albicans and Saccharomyces cerevisiae were monitored. Cyclic voltammograms (CV) showed an irreversible oxidation peak at about 750 mV that accounts for viable cells. The peak current increased at viable cell numbers ranging from 3 × 10(5) to 1.6 × 10(7)cells/ml, indicating that the amount of viable cells can be accurately quantified. To elucidate the underlying electron transfer processes, the influence of electron transfer chain (ETC) - inhibitors on the electrochemical behavior of the two organisms were investigated. Inhibition of the function of classical respiratory chain (CRC) led to a decrease in the electrochemical response, whereas the oxidation current increased when the alternative oxidase (AOX) pathway was blocked by salicylhydroxamic acid (SHA). Blocking the AOX pathway improved the electrochemical performance, suggesting an involvement in the CRC, with cytochrome c oxidase (COX) as a relevant protein complex. Mutants, in which components of COX were deleted, showed a lower electro-activity than the wild-type strain. Particularly, deletion of subunit COX5a almost completely abolished the electrochemical signal. We believe that this work can be utilized for the development of early detection assays and opens the door for new technological developments in the field of C. albicans.
尽管在该领域取得了进展,但仍缺乏用于快速检测人类病原体白色念珠菌的方法。在这方面,包括电化学传感器和生物传感器在内的生物电化学系统满足了对快速、可靠和直接微生物分析日益增长的需求。在这项研究中,研究了白色念珠菌的生物电化学特性,以用于确定生物体活力的分析系统。监测了白色念珠菌和酿酒酵母的活细胞和非活细胞的电化学响应。循环伏安法 (CV) 显示出约 750 mV 的不可逆氧化峰,该峰与活细胞有关。在活细胞数量从 3×10(5)到 1.6×10(7)个细胞/ml 的范围内,峰电流增加,表明可以准确定量活细胞的数量。为了阐明潜在的电子转移过程,研究了电子传递链 (ETC)抑制剂对两种生物体电化学行为的影响。经典呼吸链 (CRC)功能的抑制导致电化学响应降低,而当水杨羟肟酸 (SHA)阻断替代氧化酶 (AOX)途径时,氧化电流增加。阻断 AOX 途径提高了电化学性能,表明其与 CRC 有关,细胞色素 c 氧化酶 (COX)作为相关蛋白复合物。与野生型菌株相比,缺失 COX 相关蛋白的突变体表现出较低的电活性。特别是,COX5a 亚基的缺失几乎完全消除了电化学信号。我们相信这项工作可用于开发早期检测方法,并为白色念珠菌领域的新技术发展开辟了道路。