Blood waste originating from slaughterhouse activities in China is considered to be massive, and improper handling may cause epidemic diseases and environmental pollution. This research aimed to obtain a potential bacterial strain for blood protein degradation by isolating indigenous bacteria from slaughterhouses. Isolation was carried out by using dilution plate coating, while blood agar and casein plates were used to screen potential strains. Morphological, physiological, and biochemical characterizations, as well as 16S rRNA sequencing, were performed to identify the selected strain. The optimization of enzyme production was performed by using the Plackett–Burman test and response surface methodology. A strain coded NwMCC01910137 was isolated and screened to effectively degrade bovine blood protein and was identified as Bacillus subtilis. The optimum culture conditions for enzyme production included a fermentation temperature of 37.4 °C, an initial pH of 7.7, a soybean meal powder addition amount of 3.00% (w/v), a glucose level of 3.8% (w/v), a NaCl level of 0.3 g/L, a phosphate concentration of 2.5 g/L, an inoculum size of 2.5 g/L (4%), and a fermentation time of 48 h. Under the optimum condition, the strain showed enzyme activity of 166.83 U/mL. Hence, the isolated B. subtilis showed good activity in bovine blood protein degradation and has good application potential for slaughterhouse waste processing.