The United States dependence on fossil fuels has become mandatory over the past few decades. The fuel shortage during the 1970s and after Hurricane Katrina has catalyzed a need for creating alternative energy sources, improving the efficacy of these alternative energy sources, and enhancing energy sustainability. The U.S. Department of Energy has set goals to replace 30% of the liquid petroleum transportation fuel with biofuels and to replace 25% of industrial organic chemicals with biomass-derived chemicals by 2025. In the southeast United States, subterranean termites are prevalent and microbes in their gut degrade wood based materials such as cellulose which produce simple sugars that can be used to produce bioethanol. Upon seasonal change, subterranean termites undergo less enzymatic activity and wood-eating capability limiting the amount of sugars that may be produced. This limited activity sparks an interest to investigate this poorly understood phenomenon of how temperature may affect the enzymatic activity in subterranean termites’ guts. In this study, we report the development thermoresponsive biomaterial nanofiber mats containing cellulose to model cellulase activity. Using electrospinning techniques, poly(N-vinylcaprolactam) celluose fiber mats have been prepared via alkaline hydrolysis and labeled with fluorescent tags. Subterranean termites (reticulitermes species) were feed fiber mats for 10 consecutive days to assess enzyme mapping and kinetics. Fluorescent microscopy images confirmed spatial and temporal localization of cellulase enzyme throughout the termite gut upon time and temperature change. These novel high affinity enzyme detection membranes show promise towards future biofuel production.