Current methods of biomass conversion mainly utilize industrial cellular strains to upgrade feedstocks to desired bioproducts. While this approach takes advantage of the cell?s ability to grow, yields can be strongly impacted by the toxicity of the starting feedstocks, intermediate products, or final bioproduct. Toxicity can be mitigated by using cell-free bioconversion platforms. However, to date, most cell-free bioconversion platforms have been E. coli or S. cerevisiae. Cell-free bioconversion from diverse organisms? lysates can open up radically new chemical space compared to what is accessible in cells. Synvitrobio proposes to develop a cell-free fermentation platform with which to access bioproducts from non-industrial host organisms, for which E. coli- based cell-free fermentations are unsuitable. Doing so widens the range of bioproducts accessible to cell-free bioconversion. One area of demonstration is the production of toxic bioproducts, such as bacteriocins, that are traditionally produced by fermentation from non-industrial hosts. In Phase I, we will demonstrate the cell-free fermentation of nisin, a model food preservative with a $421 million market (2016). The statement of work is divided into two parts: producing nisin, and hitting cost and scale metrics. If successful, we will be well-positioned to produce nisin and other bacteriocins at scale in cell- free fermentations in a Phase II award, thereby demonstrating the unique use of the technology for cell-free bioconversion from non-E. coli hosts.