Proteases are one of the most useful enzymes in human life. These protein degrading enzymes are naturally found in many animals and plants. Plant, animal, and microbial sources are the three major sources of these enzymes. Microbial sources is preferred due to high growth rates and low nutritional requirements. In the case of microbial resources, halophilic bacteria attract gear attentions because of low risk of contamination. Waste intestinal walls of sheep, a kind of waste animal tissues, are sources of organic compounds. The release of these wastes is an environmental concern. The most effective way to deal with these wastes is their bioconversion to valuable products. In this project, the production of protease was carried out by using halophilic bacteria cultivated on waste animal tissues to create added value. First, the wastes were alkali pre-treated. After filtration, pH adjustment, and drying, the peptone powder was produced. The produced peptone powder was compared with the commercial powder in terms of moisture, total nitrogen, ash and lipid contents as well as and elemental analysis. The results revealed similarities between the commercial and produced peptone powders. In order to evaluate the performance of the produced powder, the model yeast cultured on the two powders. The results showed the similar growth rates. The powders were used as the medium components of Salinivibrio proteolyticus . The activity of the produced protease from both powders were similar. In addition, the hydrolyzed wastes were used as a substitute for the peptone powder of the medium. Different percentage of the hydrolysate were applied (2% to 90%). The results revealed the highest activity (31.94 U/ml) at 80% hydrolysate. According to the results of this study, the peptone powder produced from waste animal tissues can be used as the substitute for commercial peptone when Salinivibrio proteolyticus used for production of protease . Keywords: halophile, animal waste tissues, hydrolytic enzyme, Salinivibrio proteolyticus , protease, extremophile, hydrolysate