This analysis covers the entire biorefinery operation. The study focuses on net water consumed for the production of a unit of biofuel: blue, green, and grey water footprint. Blue water is defined as the water consumed in the biorefinery that is withdrawn from surface and ground water. Blue water footprint includes enzyme cultivation, pretreatment, hydrolysis, bioreactor, cooling system, boiler, fuel upgrading, combustor track, and on-site WWT. Grey water is defined as wastewater generated from the biorefinery and was evaluated based on the wastewater treatment plant design. Green water, defined as rainwater consumed for the production, is not required in the RDB process. Approximately 7?15 gal of water are required to produce a gallon of RDB when corn stover or non-irrigated perennial grasses, switchgrass and Miscanthus x giganteus (Miscanthus), serve as the feedstock in the contiguous United States. Bioelectricity generation from the biorefinery resulted in a net water credit, which reduced the water footprint. The life cycle grey water footprint for nitrogen is primarily from nitrogen in the feedstock production stage because no wastewater is discharged into the environment in the RDB process. Perennial grasses-based RDB production shows a promising grey water footprint, while corn stover-based RDB production has a relatively low green water footprint. Results from the study can help improve our understanding of the water sustainability of advanced biofuel technology under development. Make-up water for cooling and boiling remains a major demand in the biorefinery. The work revealed a key issue or trade-off between achieving zero liquid discharge to maximize water resource use and potentially increasing cost of fuel production. Solid waste disposal was identified as a management issue, and its inverse relationship with wastewater management could affect economic sustainability.