SBIR Phase I: Microbial production of paraxanthine, a caffeine replacement

这个小企业创新（SBIR）第一阶段项目的广泛影响是开发一种生物制造工艺来生产副黄嘌呤，作为饮料和食品中咖啡因的替代品。对于很大一部分人来说，咖啡因有副作用，包括焦虑、神经紧张和失眠。在焦虑方面，估计有4000万美国人患有焦虑症，而咖啡、能量饮料和其他饮料中的咖啡因可能会加剧这种情况。副黄嘌呤是一种活性成分，具有类似咖啡因的刺激特性，但不太可能引起焦虑和紧张。因此，副黄嘌呤可以在促进心理健康方面发挥作用。该项目旨在开发一种具有成本效益的生物方法来生产副黄嘌呤。低成本生产副黄嘌呤目前是一个重大的技术挑战。这种化合物只有在咖啡因代谢的过程中才会在人体内自然地大量存在；它不会在植物中产生超过微量的量。此外，生产副黄嘌呤的合成方法复杂、低效且昂贵。本一期项目的研究目标是结合蛋白质和微生物工程，开发一种高产高产的发酵生产副黄嘌呤的生物方法。该方法使用定向进化来选择具有改进活性的酶和微生物，从而实现实验设计-构建-测试周期的快速迭代。如果成功，这项技术不仅将提供一个高效的副黄嘌呤生产过程，而且还将建立一个新的、可扩展的平台，可以应用于改进其他食品成分、药品和工业化学品。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 (SBIR) Phase I project is the development of a biomanufacturing process to produce paraxanthine for use as a caffeine alternative in beverages and foods. For a large percentage of the population, caffeine is known to have negative side effects, including anxiety, jitteriness, and insomnia. In the case of anxiety, an estimated 40 million Americans suffer from an anxiety disorder, which may be exacerbated by caffeine in coffee, energy drinks, and other beverages. Paraxanthine is an active ingredient that has stimulatory qualities similar to caffeine but is less likely to cause anxiety and jitteriness. Thus, paraxanthine can play a role in contributing to improved mental health. The proposed project aims to develop a cost-efficient, biological method to manufacture paraxanthine. Producing paraxanthine at low cost is currently a significant technical challenge. The compound is only found naturally in substantial quantities within the human body as the result of caffeine metabolism; it is not produced in plants in above trace amounts. Furthermore, synthetic methods to produce paraxanthine are complex, inefficient, and expensive. The research goals of this Phase I project are to use a combination of protein and microbe engineering to develop a biological method to fermentatively produce paraxanthine at high yields and with high volumetric productivities. The approach uses directed evolution to select for enzymes and microbes with improved activity, enabling rapid iteration of the experimental design-build-test cycle. If successful, this technology will not only deliver an efficient paraxanthine production process, but it also establishes a novel, scalable platform that can be applied to improve upon other food ingredients, pharmaceuticals, and industrial chemicals.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.