Benthic prokaryotes include Bacteria and Archaea and dominate densities of marine benthos. They play major roles in element cycles and heterotrophic, chemoautotrophic, and phototrophic carbon production. To understand how anthropogenic disturbances and climate change might affect these processes, better estimates of prokaryotic biomasses and densities are required. Hence, I developed the ProkaBioDen database, the largest open-access database of benthic prokaryotic biomasses and densities in marine surface sediments. In total, the database comprises 1,089 georeferenced benthic prokaryotic biomass and 1,875 density records extracted from 85 and 112 studies, respectively. I identified all references applying the procedures for systematic reviews and meta analyses and report prokaryotic biomassesas g C cm⁻³ sediment, g C g⁻¹ sediment, and g C m ⁻². Density records are presented as cell cm⁻³ sediment, cell g⁻¹ sediment/ sulfide/ vent precipitate, and cell m⁻². This database should serve as reference to close sampling gaps in the future.