<p>The transition to sustainable energy has increased interest in biofuel production to reduce greenhouse gas emissions, decrease reliance on imported oil, and ensure energy resilience. This study examines the often-overlooked impact of land cost assumptions on the economic viability of biofuel production. Using a discounted cash flow techno-economic framework, we evaluated three land cost scenarios—no land costs, land rental costs, and land purchase costs—across six bioenergy feedstocks (corn, soybeans, switchgrass, miscanthus, poplar, and microalgae) and three biofuel products (corn ethanol, soybean biodiesel, and sustainable aviation fuel) at the county level for the contiguous United States. The analysis reveals substantial variation in minimum fuel selling prices due to these scenarios. High-yield crops like algae showed low sensitivity to land costs, while low-yield crops such as soybeans were highly sensitive. Geographical differences were significant, with minimum fuel selling price increases most pronounced in high-value land regions like the Corn Belt. Case studies further illustrate the influence of local productivity and land costs on economic outcomes across the United States. These findings emphasize the importance of maintaining consistent land cost assumptions in biofuel economic assessments. By quantifying the interplay between land value, crop productivity, and economic feasibility, this study provides essential insights for policymakers and stakeholders to advance sustainable energy solutions.</p>