SBIR Phase II: Production of Biocidal Enzymes for Antimicrobial Applications

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.