Study on Development of Environmental Stress Sensor

环境应力传感器的研制研究

• 批准号: 12450333
• 负责人: KUBOI Ryoichi
• 金额: $ 7.74万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12450333/
• 关键词: Stress Response; Stress Sensor System; Immobilized Liposome Electrode; Protein; Enzyme; Odor; Fragrance; Bioprocess; Process Monitoring; Conductive Polymer; スマートポリマー; 固定化リポソームクロマトグラフィー; 水性二相分配法; タンパク質; 分子間相互作用

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.

细菌细胞等生物系统在受到外界环境的各种应激时，通过从分子水平到细胞水平不同层次的组成元素之间的自组织和协同相互作用，并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，需要开发“应激传感器”，以识别各种构成元素（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。本研究开发了应激传感器系统，用于连续原位监测生物膜的状态和细胞表面的环境应激源（应激蛋白、气味/香味分子等）。围绕生物膜的动态应激响应，开发了一系列模仿多种生物功能的传感器，此外，还开发了识别由应激引起的气味/香味分子等环境应激源的方法。除了使用固定化脂质体色谱（ILC）评估蛋白质-脂质体相互作用之外，还建立了评估模型生物膜（脂质体）之间相互作用的方法，还开发了脂质体固定电极，并通过检测与应激反应功能相关的各种状态（即蛋白质的解折叠和重折叠， 基于上述结果，开发了监测和控制应激响应性生物过程的方法，并被证明可应用于应激响应性生物过程的生产和分离。