SBIR Phase I: Platform for rapid assessment of hydrolytic enzyme activity to optimize their design and use

这项小企业创新研究（SBIR）第一阶段项目的广泛影响将是加速和改进酶产品的开发和优化。该项目将开发一个平台，减少与研究相关的成本和延迟。这些酶产品将在一系列应用中提高成本、安全性和可持续性，如生物质加工、动物营养、固体和液体废物管理、家庭和工业清洁剂以及酶替代疗法。该解决方案将支持100亿美元酶市场的增长。拟议中的项目将通过减少开发新的工业生物催化剂的障碍，推进基于荧光的酶活性传感平台，通过开发模块化酶活性探针，使工业和专业市场的水解酶快速设计成为可能。拟议的研究和开发解决的技术障碍是：(1)确定相对于现有表征技术的性能指标（检测限制，速度，准确性），(2)确定传感器制备方法对传感器性能的影响，以及(3)开发用于现场使用的强大读取器。拟议的研究和开发的目标是：(1)为最终用户提供当前分析的奇偶数据，以产生对我们方法的信心，(2)标准化生产协议，以及(3)提供便宜（3000美元）并且可以承受野外粗暴处理的手持读取器。在高层次上，这些目标将通过(1)在已经具有严格表征的候选酶面板上测试我们的传感器（以建立性能指标），(2)使用设计的实验来筛选传感器制备的过程空间和对传感器性能属性的模型影响，以及(3)使用设计-构建-测试工程框架来测试读取器组件和布局。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact of this Small Business Innovation Research (SBIR) Phase I project will be to accelerate and improve the development and optimization of enzyme products. This project will develop a platform that reduces costs and delays associated with research. These enzyme products will improve the cost, safety, and sustainability in a range of applications, such as biomass processing, animal nutrition, solid and liquid waste management, household and industrial cleaners, and enzyme replacement therapies. This solution will support the growth of the $10 billion enzyme market.The proposed project will advance a fluorescence-based enzyme activity-sensing platform, by reducing barriers to development of new industrial biocatalysts, enabling the rapid design of hydrolytic enzymes for industrial and specialty markets through the development of modular enzyme activity probes. The technical hurdles addressed by the proposed research and development are (1) determining performance metrics (limits of detection, speed, accuracy) relative to existing characterization techniques, (2) determining effects of sensor preparation methods on sensor performance, and (3) developing a robust reader for field use. The goals of the proposed research and development are to (1) provide parity data to current assays for end-users to engender confidence in our approach, (2) standardize production protocol, and (3) provide hand-held reader that is inexpensive ($3000) and can withstand rough handling in field. At a high level, these goals will be achieved by (1) testing our sensors on a panel of candidate enzymes that have rigorous characterization already (to establish performance metrics), (2) using designed experiments to screen process space of sensor preparation and model effect on sensor performance attributes, and (3) using a design-build-test engineering framework to test reader components and layout.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.