SBIR Phase I: A Biocide/Biocide-Resistant System for Microbial Contamination Control in Biomanufacturing

这个小型企业创新研究（SBIR）项目的更广泛的影响/商业潜力是开发能够实现稳健、无污染、连续的生物制造的技术。虽然生物制造可能是技术上最复杂和最有利可图的行业之一，但潜在的生产过程，批量发酵，几十年来没有改变。理论上，连续发酵可以降低操作成本，提高生产能力。然而，由于污染的高风险，连续发酵被认为是不可靠的，很少使用。所提出的技术解决了这一问题，并使无污染的连续发酵成为可能。连续发酵的成功实施促进了自动化生产，为未来连续产品纯化和回收的技术发展铺平了道路。降低运营成本对于生物基商品化学品（如生物塑料）的经济可行性至关重要，因为它们的市场需求主要取决于它们与石油衍生产品的价格竞争力。拟议的技术还消除了抗生素在发酵中的使用，这反过来又减轻了抗生素的滥用，并促进了积极的环境和健康影响。这个SBIR一期项目的智力优势在于探索在连续生物制造过程中使用杀菌剂/抗杀菌剂系统来预防和处理微生物污染的可行性。生物制造过程中的微生物污染是一个主要问题，因为它会导致生产力、时间和金钱的损失。此外，它将生产方式限制为缓慢分批发酵，禁止实施连续发酵。该技术解决了污染问题，因为杀菌剂可以抑制入侵的微生物和病毒污染物，而抗杀菌剂酶可以保护加工宿主。拟议的第一阶段研究将通过使用经典分子生物学和合成生物学技术来解决技术挑战。第一个目标是证明所提出的技术将为产生生物塑料的生物体提供抗杀菌剂，同时抑制污染物的生长。第二个目标是证明所提出的技术可用于防止和处理连续发酵罐中的污染。最终目标是微调杀菌剂/抗杀菌剂系统，以证明该技术不会对连续发酵罐的产量和产品质量产生不利影响。如果成功，未来的第二阶段开发将在试点和工业规模的反应堆中进行，以进一步优化技术。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/commercial potential of this Small Business Innovation Research (SBIR) project is to develop technology that will enable robust, contamination-free, continuous biomanufacturing. While biomanufacturing may be one of the most technologically complex and profitable industries, the underlying production process, batch fermentation, has not changed in decades. In theory, continuous fermentation would decrease operational cost and increase production capacity. However, owing to the high risk of contamination, continuous fermentation is considered unreliable and is rarely used. The proposed technology solves this issue and enables contamination-free, continuous fermentation. Successful implementation of continuous fermentation facilitates automated production and paves the way for future technological development in continuous product purification and recovery. The reduced operational cost is essential for the economic viability of bio-based commodity chemicals such as bioplastics, as their market demand depends primarily on their price competitiveness with petroleum-derived counterparts. The proposed technology also eliminates the use of antibiotics in fermentation, which in turn alleviates antibiotic misuse and promotes a positive environmental and health impact. The intellectual merit of this SBIR Phase I project is to explore the feasibility of using a biocide/biocide-resistant system to prevent and treat microbial contamination during continuous biomanufacturing. Microbial contamination in biomanufacturing processes is a major concern, as it results in loss of productivity, time, and money. In addition, it restricts the production mode to slow batch fermentation, and prohibits the implementation of continuous fermentation. The proposed technology solves the contamination problem, as a biocide will inhibit the invading microbial and viral contaminants and a biocide-resistant enzyme will protect the processing hosts. The proposed Phase I research will address the technical challenges by using classical molecular biology and synthetic biology techniques. The first objective is to demonstrate that the proposed technology will provide biocide resistance to bioplastics-producing organisms while inhibiting the outgrowth of contaminants. The second objective is to show that the proposed technology can be used to prevent and treat contamination in continuous fermenters. The final objective is to fine-tune the biocide/biocide-resistant system to demonstrate the technology will not adversely affect the production yield and product quality in continuous fermenters. If successful, future Phase II development will be conducted in pilot and industrial-scale reactors to further optimize the technology.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.