The Gulf of Mexico (GoM) is a complex oceanic basin with a maximum depth of 4000 m. It is a complex hydrodynamic system formed by different water masses with distinctive physical and biological characteristics that shape its rich biodiversity. In this study, as a contribution to better understanding the microbial communities inhabiting the meso- and bathypelagic zones of the Mexican Exclusive Economic Zone (EEZ) of the GoM, an extensive set of seawater samples was collected at three depths (350-3700 m) during three oceanographic cruises. The V4-16S rRNA gene analysis identified Pseudomonadota (27.1 ± 9.8%) and Nitrosopumilales (26.4 ± 2.3%) as the dominant bacterial and archaeal members, respectively. The depth, salinity, and apparent oxygen utilization were key environmental drivers, which explained 35% of the community variability. The mesopelagic zone presented a more homogeneous structure characterized by a nitrifier community, while the bathypelagic was more heterogeneous, with hydrocarbon-degrading bacteria and methanogens serving as the key players. This study is the first to report the archaeal community in the deeper waters of the Mexican EEZ of the GoM, playing crucial roles in the nitrogen and carbon cycles, highlighting the region's ecological complexity and the need for further research to understand the broader biogeochemical implications of these processes.