Study on Development of Environmental Stress Sensor

细菌等生物系统通过从分子水平到细胞水平的不同水平的组成元素之间的自组织和协同作用，以及当它们受到来自外部环境的各种压力时，通过形成高度有序的结构来诱导应激反应功能。在利用和控制细菌细胞和生物系统的应激反应功能的“应激反应生物过程”中，为了识别各种构成要素(即细胞、酶、蛋白质、细胞膜、生物分子)和应激源的应激状态并对其进行监测，需要研制一种“应激传感器”。在本研究中，开发了一种应力传感器系统，用于连续在线监测生物膜和细胞表面的环境应激源(应激蛋白质、气味/芳香分子等)的状态。针对生物膜的动态应激反应，开发了一系列模拟多种生物功能的传感器，并进一步发展了识别应激诱导的气味/芳香分子等环境应激源的方法。建立了模型生物膜(脂质体)之间相互作用的评价方法，建立了利用固定化脂质体层析(ILC)评价蛋白质-脂质体相互作用的方法，建立了脂质体固定电极，并通过检测与应激反应功能相关的各种状态(即蛋白质的展开和折叠、蛋白质转位和膜融合)来区分蛋白质构象变化的方法。在此基础上，建立了应激反应生物过程的监测和控制方法，并将其应用于应激反应生物过程的生产和分离。

The biological systems such as bacterial cells are known to induce the stress-response functions through the self-organizing and cooperative interaction between constituting elements at different levels from molecular level to cell level and, also, through the formation of highly-ordered structures when they are exposed to a variety of stresses from the external environment. In the "stress responsive bioprocess" utilizing and controlling the stress-response functions of bacterial cells and biological systems, the development of "a stress sensor" are needed in order to recognize the stress state of various constituting elements (i.e. cells, enzyme, protein, cell membrane, biomolecules) and stressors and to monitor it. In this study, the stress sensor system was developed for the in situ monitoring of the state of the biological membrane and the environmental stressors (stressed proteins, odor /fragrance molecules and so on) on the cell surface continuously. A series of sensors mimicking a variety of biological functions were developed focusing on the dynamic stress-response of biological membrane and, furthermore, the method to recognize the environmental stressors such as odor/fragrance molecules, induced by the stress, was developed. The method to evaluate the interaction between the model biological membranes (liposomes) was established in addition to the protein-liposome interaction by using the immobilized liposome chromatography (ILC), The liposome immobilized electrode was also developed and the method to differentiate the conformational change of proteins by detecting various state relating to the stress response functions (i.e. unfolding and refolding of protein, protein translocation and membrane fusion by model biological membrane).Based on the above results, the method to monitor and control the stress-responsive bioprocess was developed and was shown to be applied for the stress-responsive bioprocess for production and separation.