SBIR Phase II: Production of Biocidal Enzymes for Antimicrobial Applications

小企业创新研究（SBIR）第二阶段的更广泛影响/商业潜力是在日常产品中用可持续的替代品取代化学防腐剂。到目前为止，制造商几乎没有替代品来防止含水产品的微生物污染。然而，酶是可生物降解的，穿透皮肤的风险很低，使其成为传统化学防腐剂的一种有吸引力的替代品。 拟议的项目将通过开发可扩展的制造工艺，促进可持续抗菌酶作为化学防腐剂替代品的转化。拟议的SBIR第二阶段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。该项目的重点是提高活性生物杀灭酶的产量并增强其抗菌性能。这些酶对细菌和酵母菌有很高的毒性，使它们成为化学防腐剂的有吸引力的替代品，但对大规模生产造成了挑战。该项目结合了一套化学和遗传开关，以在生产过程中紧密调节酶的杀生物活性，从而在微生物宿主生物体中生产过程中实现高酶滴度。研究目标有两个方面：1）优化生产菌株和遗传学，以过表达杀生物酶，2）优化最终酶产品的下游加工，以最大限度地提高杀生物活性。这些目标将通过新型定向进化平台和已建立的应变工程基础设施的组合来实现。该奖项反映了NSF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.