Filamentous bacterial belonged to
Streptomyces
species were novel drug source for medical and industrial applications. However, the detailed identification of
Streptomyces
species from Saudi Arabian extreme environment for the identification novel drug source for medical and industrial applications were rarely studied. The
Streptomyces
strain Al-Dhabi-2 obtained from the thermophilic region kingdom of Saudi Arabia, exhibited antimicrobial potentials against the pathogenic microorganism were characterized. Biochemical and phylogenetic analysis confirmed that the strain was closely associated to the
Streptomyces
species. The chromatogram of GC-MS analysis of this ethyl acetate extract (EA) had diverse of chemical compounds namely benzene acetic acid (7.81%), acetic acid, methoxy-, 2-phenylethyl ester (6.01%) were the major compounds. EA of Al-Dhabi-2 showed inhibition zone ranged from 14 to 25 mm at 5 mg/well concentration against the tested microbial pathogens. Results revealed that the significant MIC values were observed against
B. cereus,
and
E. faecalis
by (less than 39 μg/ml) and against
S. agalactiae
with (78 μg/ml). Minimum inhibitory concentrations (MIC) for fungi: were also reported against
Cryptococcus neoformans
and
Trichophyton mentagrophytes
by (156 μg/ml), whilst
Candida albicans
and
Aspergillus niger
by (312 μg/ml). Results of this study showed that thermophilic actinobacteria could be promise source in the context of searching for unique antimicrobial agents with novel properties.