Aim: To identify the penicillin resistance organisms in soil sample collected from the hospital area. Methods: Bacteria were grown on nutrient agar medium and sub cultured in nutrient broth with different concentrations of Penicillin. The resistance organisms were characterized by 16s rRNA gene sequencing and blast analysis. Results: A total of 21 cultural were identified as penicillin resistance organisms. Out of 21 cultures we have characterized four cultures and identified as Pseudomonas putida, Pseudomonas fluorescens, Comamonas sp. KBB4, Bacterium SM2-6. Conclusion: Pseudomonas putida, Pseudomonas fluorescens, Comamonas sp. KBB4, Bacterium SM2-6. Four cultures were identified as penicillin resistance organisms. Remaining 17 cultures need to characterize.