Environmental baseline information is necessary in order to assess the potential environmental impact of future manganese-nodule mining on the deep-seabed ecosystem. Total ATP (T-ATP), dissolved ATP (D-ATP) and particulate ATP (P-ATP) were measured to estimate total microbial biomass and to elucidate their vertical distribution patterns in the seabed of KODOS (Korea Deep Ocean Study) area, northeast equatorial Pacific Ocean. Within the upper 6 cm depth of sediment, the concentrations of T-ATP, D-ATP and P-ATP ranged from 4.4 to 40.6, from 0.6 to 16.1, and from 3.0 to 29.2 ng/g dry sediment, respectively. Approximately 84% of T-ATP, 81% of D-ATP, and 74% of P-ATP were present within the topmost 2 cm depth of sediment, and the distributions of ATP were well correlated with water content in the sediment. These results indicate that the distribution of total microbial biomass was largely determined by the supply of organic matter from surface water column. Fine-scale vertical variations of ATP were detected within 1-cm thick veneer of the sediment samples collected by multiple corer, while no apparent vertical changes were observed in the box-cored samples. It is evident that the box-core samples were disturbed extensively during sampling, which suggests that the multiple corer is a more appropriate sampling gear for measuring fine-scale vertical distribution pattern of ATP within thin sediment veneer. Overall results suggest that the concentrations of ATP, given their clear changes in vertical distribution pattern within 6 cm depth of sediment, are a suitable environmental baseline parameter in evaluating the variations of benthic microbial biomass that are likely to be caused by deep-seabed mining operation.