Deciphering the biological resources across the Saudi niches is highly recommended for the prosperity. To this end, the aim of the current work was to isolate thermophilic bacteria from unexplored areas of Al-Ahsa region, and investigate their phenotypic characteristics. Three soil samples were collected from different desert sites of Al-Ahsa region. Thermophilic bacteria were isolated directly for soil samples into Thermus medium broth as a standard method. Single colonies of the actively growing bacterial isolates were preserved in 20% glycerol then kept at -80°C. The isolates were screened for production of thermostable enzymes using the commercially available kit API20E strip (bioMerieux, Marcy l’Etoile, France). Incubation were carried out at 50°C. It can be concluded that thermophilic bacteria in Al-Ahsa region harbor novel thermostable enzymes that might have biotechnological applications, in future.