This study was aimed to isolate and identify of Klebsiella pneumonia and testing the production of Arginine deiminase (ADI) purified from Klebsiella pneumonia in vitro, 30 samples were collected from sheep infected with respiratory infections from veterinary clinics and hospitals from October to April 2024 then transported by transport media to the laboratory for culture and microbial examine. Out of the 21 samples, 13 (61.9%) showed bacterial growth, with the majority being Gram-negative bacteria. The remaining 8 samples (38%) were gram-positive. The analysis revealed that Klebsiella pneumoniae was the most identified bacterial strain. Out of the total bacteria used, 7 (53.8%) were identified as gram-negative bacteria, while the remaining were of different types. This study identified the enzyme arginine deiminase from 7 clinical isolates of Klebsiella pneumonia whereby the enzyme was extracted and purified. The objective was to quantify K. pneumonia for this enzyme production capacity to measure its overall capability on subsequent phases via screening. This screening method was based on the measurement of the precise arginine deiminase activity using spectrophotometric assay. The specific activity of purified enzyme was 10 U/min with approximate molecular weight 120 kDa and the isolate K. pneumonia N7 was found most effective in production of the enzyme.