SBIR Phase II: Commercialization of Synthetic Metabolic Valves

这一小型企业创新研究(SBIR)第二阶段项目如果成功，其更广泛的影响/商业潜力将是展示该技术的潜力，该技术将大幅降低目前与生物产品生产相关的成本和风险，从而实现超低成本产品开发。代谢工程领域历史上一直局限于根据简化的模型和基本的体外生化原理来预测复杂生物系统在体内的行为。在许多情况下，事实证明，将一条具有良好特性的生产途径整合到活的宿主中并平衡生物量增长和生产的复杂要求比预期的要困难得多。正在开发的技术是此类技术中的第一个，真正可扩展的高通量代谢工程平台，能够快速开发微生物生产菌株。开发成本的显著降低使生产大量特殊产品成为可能，否则这些产品将无法获得可接受的开发资本内部回报。这个SBIR二期项目将开发一个高通量代谢工程平台，使微生物生产菌株能够快速开发。该平台在目前体内和体外生物生产方法之间架起了一座桥梁，依赖于标准化两阶段生物过程中活性代谢网络的动态最小化。代谢网络是高度相互联系的，其中每个代谢物和/或酶可以与无穷无尽的其他代谢物和/或酶相互作用。这种组合的复杂性导致了巨大的潜在设计空间，这对于开发标准化设计原则所需的各种系统实验来说是棘手的。尽管在读取和写入DNA、高通量DNA组装和微生物菌株构建方法方面取得了显著进展并降低了成本，但在解决如此大的生物设计空间方面的全球挑战仍然存在。动态新陈代谢网络最小化不仅使设计空间的复杂性大大降低，而且还提供了对环境条件稳健的菌株。健壮性带来了可预测的可伸缩性，从高通量的小规模筛选或“微发酵”到完全仪表化的生物反应器。该项目将把可预测菌株性能的验证从高通量微发酵扩展到中试规模发酵。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project, if successful, will be to demonstrate the potential of the technology to dramatically reduce the cost and risk currently associated with production of bio-based products, enabling ultra-low cost product development. The field of metabolic engineering historically has been limited in predicting the behavior of complex biological systems in vivo from simplified models and basic in vitro biochemical principles. In many cases, it has proven much more difficult than expected to integrate a well characterized production pathway into a living host and balance the complex requirements of both biomass growth and production. The technology under development is a first of its kind, truly scalable, high-throughput metabolic engineering platform enabling the rapid development of microbial production strains. This significant reduction in development cost enables the possibility to produce numerous specialty products that would otherwise not have acceptable internal return on development capital. This SBIR Phase II project will develop a high throughput metabolic engineering platform that enables the rapid development of microbial production strains. The platform, which bridges a gap between current in vivo and in vitro bio-production approaches, relies on the dynamic minimization of the active metabolic network in the context of a standardized two-stage bioprocess. Metabolic networks are highly interconnected wherein each metabolite and/or enzyme can interact with endless others. This combinatorial complexity results in a huge potential design space, which is intractable to the kinds of systematic experimentation required for the development of standardized design principles. The global challenges in addressing such a large biological design space have persisted, despite the dramatic advances in, and decreased costs of, reading and writing DNA, high-throughput DNA assembly, and microbial strain construction approaches. Dynamic metabolic network minimization not only results in a design space with greatly reduced complexity, but also provides strains that are robust to environmental conditions. Robustness leads to predictable scalability from high-throughput small-scale screens or "microfermentations" to fully instrumented bioreactors. This project will extend the validation of predictable strain performance from high-throughput microfermentation to pilot scale fermentation.