The aim of the study was to identify new sources of protease and a protease producing bacteria was isolated from slaughter house soil and identified as Pseudomonas species. The protease production efficiency of the organism was measured with different environmental and nutritional parameters. Optimization of the fermentation medium for maximum protease production was carried out. The culture condition like temperature, pH, incubation time, carbon and nitrogen sources were optimized. The optimum condition found for protease production was 30°C at pH 7 in the medium for 48 hour. Carbon source casein and nitrogen source beef extract stimulated the production of protease. Protease production was higher with immobilized cells. Pseudomonas species can be used profitably for the large scale production of protease to meet the present day demand of the industrial sector.DOI: http://dx.doi.org/10.3329/icpj.v2i1.12870 International Current Pharmaceutical Journal 2012, 2(1): 1-6