The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project, if successful, will be the development of a microbial process for the economical and sustainable production of a highly potent natural acaricide, which is a pesticide that kills mites and ticks. 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 and safe acaricide; however, its commercialization has been hampered by a high cost of production. The aim is to develop an alternative manufacturing process for biosynthetic production enabling the cost reductions required to effectively penetrate the $1.6 B acaricide market. Because the target is GRAS and because it has been used extensively as a food ingredient for decades, there is a compelling safety benefit combined with its potent efficacy, which may spur increased spraying in public areas and private residences. Overall, this project will provide a new sustainable, cost-effective production route, thereby enabling acaricide commercialization. <br/><br/>This SBIR Phase II project will lead to sustainable, scalable, and economical access to a highly potent natural acaricide. A commercial fermentation process will be developed by employing advanced metabolic engineering and protein engineering approaches for improving strain and enzyme performance. Achieving these production metrics will enable formulation and commercialization of various acaricidal products, including yard/area sprays, which will allow better control of tick populations and halt the spread of tick-borne diseases such as Lyme disease. In addition, this work will significantly advance the understanding of producing complex plant natural ingredients, thus providing economical and scalable commercial access to a wide array of compounds with significant potential benefit.