Industries of food, pharmaceutical, agricultural and medical have been taking advantage of using Bacillus sp. because of their wide range of physiological characteristics and ability to produce enzymes and other metabolites. Bacillus sp. are attractive industrial microorganisms recognized as generally recognized as safe, which have high growth rates leading to shorter fermentation times and possess the ability to secrete extracellular proteins. Production of protease was carried out generally using submerged fermentation. It is advantageous than other methods due to its consistent enzyme production with defined medium, better process conditions and improved downstream processing. Therefore, current study was conducted with the main objectives of isolation of bacterial strain producing protease enzyme and optimization of submerged fermentation parameters for enhanced production of protease enzyme. Slaughter house waste from local market of Chikkaballapura were collected and screened for protease producing bacterial strains on skimmed milk agar plates. Protease positive bacterial strains were identified and recorded based on the clear zone formation around the bacterial growth. Protease producing bacteria was grown on the optimized media in submerged fermentation method. Optimization of cultural conditions like incubation period, pH, temperature, carbon & nitrogen sources was carried out. Folin Ciocalteu method of enzyme assay was followed and activity was expressed in U/mL. Study findings delineated that newly isolated Bacillus subtilis produced extrasellar protease optimally when it was cultured at 50ºC, pH 8.0 for 36 hours under submerged fermentation. Furthermore, newly isolated B. subtilis utilized fructose and yeast extract as carbon and nitrogen sources respectively for optimal production. In conclusion, newly isolated B. subtilis in this study has potential industrial biotechnological applications. Therefore, large scale production studies utilizing newly isolated B. subtilis could be recommended to carry out in order to exploit for industrial applications.