Spore-Based Biosensing Systems: A Stabilized Dormant-Active Approach to Whole-Cell Biosensors

化学系的分析和表面化学项目将支持肯塔基大学Sylvia Daunert教授的研究项目。Daunert教授的研究项目专注于全细胞传感系统的开发。全细胞传感系统是理想的分析工具，在生物医学分析和环境监测，由于其灵敏度，选择性和快速。如果能够有效地用于现场应用，它们的潜力将进一步增强。在这方面，需要改进全细胞生物传感系统的保质期和可运输性的方法，并允许它们适应用于现场分析的小型化系统。为此目的，Daunert教授和她的学生将开发一种基于利用细菌孢子长期维持细菌感应细胞的活力和活性的方法，并将这种保存的感应细胞整合到便携式系统中进行现场感应。选择细菌孢子作为全细胞传感器的保护成分，因为它们是一种静止的生命形式，对极端环境条件表现出长期的抵抗力，并且在适当的刺激下能够恢复完全的代谢活动。具体来说，Daunert教授和她的学生将利用孢子形成细菌开发用于各种目标分析的基因工程全细胞传感系统。然后，它们会产生孢子，作为保存、储存和运输感应细胞的手段。随后，他们将把这些休眠传感器整合到一个小型紧凑的圆盘状微流控平台中。此外，他们将固定在纸条作为一个简单的即用型配置。这两种系统都可以被运送到野外，在那里孢子可以发芽为营养细胞并用于传感。Daunert教授设想，这种新策略可以扩大全细胞生物传感器在现场分析中的应用，不仅可以在温和的环境中使用，还可以在以前无法使用的环境和应用中使用。这些包括极端环境，如沙漠、极地和太空，以及用于生物恐怖主义的化学战剂的遥感。这项工作对发展中国家的影响可能很大，因为这些国家需要快速、简单和廉价的分析工具来监测人类健康和环境，而不利的气候条件和分配和储存设施不足往往会造成限制。该项目将为学生提供在具有高社会影响力的前沿多学科研究领域的优秀培训机会。

The Analytical and Surface Chemistry program in the Division of Chemistry will support the research program of Prof. Sylvia Daunert of the University of Kentucky. The research program of Prof. Daunert focuses on the development of whole-cell sensing systems. Whole-cell sensing systems are ideal analytical tools in biomedical analysis as well as in environmental monitoring due to their sensitivity, selectivity, and rapidity. Their potential would further be enhanced if they could be used effectively for on-site applications. In that regard, there is a need for methods that improve the shelf-life and transportability of whole-cell biosensing systems, and allow for their adaptation to miniaturized systems for field analysis. To that end, Prof. Daunert and her students will develop a method based on the use of bacterial spores for the long-term maintenance of the viability and activity of bacterial sensing cells, and integrate such preserved sensor cells into portable systems for on-site sensing. Bacterial spores were chosen as the protective component of whole-cell sensors since they are a resting form of life that exhibit long-term resistance towards extreme environmental conditions, and are able to resume full metabolic activity when properly stimulated. Specifically, Prof. Daunert and her students will develop genetically engineered whole-cell sensing systems for various target analytes using spore-forming bacteria. They will then produce spores as means of preservation, storage and transport of the sensing cells. Subsequently, they will incorporate these dormant sensors into a miniaturized compact disk-like microfluidic platform. Additionally, they will immobilize them on paper strips as a simple ready-to-use configuration. Both systems can be transported to the field, where the spores can be germinated to vegetative cells and employed for sensing. Prof. Daunert envisions that this new strategy could expand the use of whole-cell biosensors for on-site analysis, not only in mild environments, but also in environments and applications in which they could not be employed previously. These include extreme environments, such as deserts, polar regions and space, as well as the remote sensing of chemical warfare agents used for bioterrorism. The impact of this work could be strong in developing countries, where rapid, simple and inexpensive analytical tools for monitoring human health and the environment are needed, and limitations are often posed by unfavorable climate conditions and inadequate distribution and storage facilities. The project will provide excellent training opportunities to students in a cutting edge multidisciplinary research area of high societal impact.