The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project will be to reduce the incidence of Lyme Disease through the biomanufacturing of a novel natural acaricide. New cases of Lyme Disease have grown by nearly 50% over the past decade while the existing synthetic acaricides are dwindling in use due to regulatory and consumer safety concerns. The CDC and USDA have begun to champion a highly effective natural acaricide extracted from grapefruit. This target molecule is a GRAS-approved natural product, which has been used extensively as a food ingredient for decades. It is thought that this compelling safety benefit combined with potent efficacy will spur increased spraying in public areas and private residences. However, the cost of producing this natural acaricide has been prohibitive, and there is an opportunity to develop alternative sustainable production technologies.<br/><br/>This SBIR Phase I project proposes to develop a microbial process for the economical and sustainable production of a highly potent natural acaricide. Increasing wariness of synthetic insecticides combined with the need to prevent tick-borne illnesses creates a tremendous opportunity for natural acaricides. The project's terpene target has long been known as a highly effective acaricide; however, its commercialization has been hampered by a high cost of production. The aim is to develop an alternative fermentation process for biosynthetic production enabling the cost reductions required to effectively penetrate the acaricide market. The main objective for this project is to increase titers by an order of magnitude. This will be accomplished by employing established and novel metabolic and protein engineering approaches. Overall, this project will provide a new sustainable, cost-effective production route, thereby enabling acaricide commercialization.