SBIR Phase II: Production of Biocidal Enzymes for Antimicrobial Applications

这项小企业创新研究（SBIR）第二阶段的更广泛影响/商业潜力是用可持续替代品取代日常产品中的化学防腐剂。迄今为止，制造商几乎没有防腐剂的替代品，这些防腐剂对防止含水产品的微生物污染至关重要。然而，酶是可生物降解的，穿透皮肤的风险很低，这使它们成为传统化学防腐剂的一个有吸引力的替代品。拟议的项目将通过开发可扩展的制造工艺，推进可持续抗菌酶作为化学防腐剂替代品的翻译。拟议的SBIR二期项目将推进生物杀灭酶的转化，以取代消费品中的化学防腐剂。本项目的重点是改进活性杀菌酶的生产和增强其抗菌性能。这些酶对细菌和酵母剧毒，使它们成为化学防腐剂的有吸引力的替代品，但对大规模生产构成挑战。该项目采用了一套化学和遗传开关，以在生产过程中严格调节酶的生物杀灭活性，从而在微生物宿主生物生产过程中实现高酶滴度。研究目标有两个：1)优化过表达杀菌酶的生产菌株和遗传，2)优化最终酶产品的下游加工，以最大限度地提高杀菌酶的活性。这些目标将通过结合新的定向演化平台和已建立的应变工程基础设施来实现。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) Phase II is to replace chemical preservatives with sustainable alternatives in everyday products. To date, manufacturers have had few alternatives to the preservatives critical for preventing microbial contamination of water-containing products. However, enzymes are biodegradable and have a low risk of penetrating the skin, making them an appealing alternative to traditional chemical preservatives. The proposed project will advance the translation of sustainable antimicrobial enzymes as replacements for chemical preservatives by developing a scalable manufacturing process. The proposed SBIR Phase II project will advance the translation of biocidal enzymes to replace chemical preservatives in consumer products. The focus of this project is improving production of the active biocidal enzyme and potentiating its antimicrobial properties. These enzymes are highly toxic to bacterial and yeast, making them attractive alternatives for chemical preservatives, but creating a challenge for manufacturing at scale. This project incorporates a suite of chemical and genetic switches to tightly modulate the enzymes' biocidal activity during production to allow for high enzyme titers during production in a microbial host organism. The research objectives are two-fold: 1) optimize production strains and genetics for overexpression of the biocidal enzyme, and 2) optimize downstream processing of the final enzyme product to maximize biocidal activity. These objectives will be fulfilled using a combination of a novel directed evolution platform and established strain engineering infrastructure.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.