Abstract: Background and Objective: Hydrocarbon contamination is one of the important environmental problems resulting from activities related to the production and use of petrochemicals. The aim of current study was to isolate, characterize and evaluate the potential for treating petroleum hydrocarbons using actinomycetes and bacterial strains isolated from petroleum-contaminated soil. Materials and Methods: Two Pseudomonas isolates and three Actinomycetes isolates were isolated from contaminated soil and then identified using 16S rDNA gene sequencing. These isolates tested for high degradation capacity of petroleum oil. Results: Actinomycetes and bacterial isolates that showed biodegradation capacities were molecular identified as Streptomyces sampsonii, Streptomyces orinoci, Streptomyces ferralities, Pseudomonas xanthomarina and Pseudomonas proteolytica. These strains were subjected for further study to measure the quantitative degradation of saturated and polycyclic aromatic hydrocarbons (PAHs). Streptomyces ferralities showed the highest degradation percent of saturated hydrocarbons, reached 82.106 and 81.672 for normal-paraffin and iso-paraffin, respectively. Maximum biodegradation percent of polycyclic aromatic hydrocarbons (PAHs) was 54.798 by using Streptomyces sampsonii. Conclusion: The observed PAHs activities of these actinomycetes strains suggested their potential usage as an alternative sources of hydrocarbon chemical degradation in petroleum industry.