Permafrost degradation creates hydrologically distinct landscapes, including dry soils with enhanced drainage and wet soils with poor drainage. In Fairbanks (Alaska, USA), we compared the active layer of dry and wet degraded permafrost landscapes with that of a non-degraded intact permafrost landscape to assess how hydrological conditions shape microbial community composition, extracellular enzyme activities, and microbial extracellular polymeric substances (EPS). Based on 16S rRNA sequencing, the alpha diversity of the prokaryotic community was significantly lower in the dry site compared to the wet and intact sites, with soil organic carbon (SOC) and pH identified as the primary environmental drivers. The active layer of the dry site was dominated by Thermoleophilia and Acidobacteriae, whereas the wet site was dominated by Gammaproteobacteria and Gemmatimonadetes. The relative proportions of most bacterial classes differed significantly across depths in the dry and wet sites, while the intact site exhibited less vertical variation. Hydrolytic enzyme activities were significantly higher in the topsoil across all sites, whereas oxidative enzyme activities showed relatively uniform patterns with depth but generally exceeded hydrolytic activities. EPS content varied among sites and depths, with the highest EPS-sugar content observed in the wet site overall, whereas topsoil EPS-sugar and EPS-protein contents were highest in the wet site, followed by the dry and intact sites, and subsoil EPS content was highest in the intact site. This study demonstrates that hydrological shifts in degraded permafrost soils shape microbial community structure, enzymatic activity patterns, and EPS content, with potential implications for SOC loss and stabilization.