Engineered Strain for Bioconversion of Food Waste to Fuels

The proposed strategy in this STTR proposal for bioconversion of food waste to butanol uses the well-described Clostridium acetobutylicum organism to convert the carbohydrate to the fuel molecule butanol. Based on our teams many years of work with butanol formation and genetic studies of the organism and the advancement of recently developed genetic methods, the team will develop a set of genetic tools that show proof of principle for demonstration of 5 or more native regulatory elements with differential responses that can be used as orthogonal controls of expression depending on environmental conditions or substances present. These differential controls will be related to the presence of various sugars, amino acids, or redox conditions. We will show their induction in cultures with promoter and regulator controlled expression of an easily detectable reporter. We will also demonstrate the use of these control systems to express other genes that will enhance pathways for feedstock utilization or improved metabolite distribution toward the product. These experiments will prepare a standard method and toolkit for manipulation of other microbes for improved use of food waste and convert a higher proportion to useful fuels and thus reduce waste costs while generating useful products for the military.