Purification and Characterization of Non-hemolytic Fibrinolytic Protease From Newly Isolated Proteus penneri SP 20.

The most common cause of cardiovascular diseases which impact blood arteries is thrombosis. Current medications and protein-based treatments are examples of thrombolytic agents. Finding new enzymatic sources is essential in health difficulties because all of these agents have unfavorable side effects. By taking this fact into account, this study presents the purification and biochemical characterization of a fibrinolytic metalloprotease from Proteus penneri SP-20 (AB909494) as a new isolate. The enzyme was successfully purified using DEAE-cellulose anion exchange chromatography at a recovery rate of 58.73% and at 33.38 purification fold. The SDS-PAGE examination yielded a molecular mass of 16 kDa. The isolated enzyme was most active at 40 °C and pH 6.0. The purified enzyme maximal velocity (Vmax) and Michaelis constant (Km) were determined to be 6 × 10-4 mol/L/min and 2.07 mg/mL, respectively. Phenanthroline, EDTA, Hg⁺, Ag⁺, Pb⁺, Li⁺, and Co⁺ all significantly lowered enzyme activity (≥ 70%), but Mn⁺, Fe⁺, Triton X-100, and urea markedly increased it. The enzyme showed considerable stability in a variety of laundry detergents and stability in toluene and hexane. The purified enzyme was highlighted as a potential safe thrombolytic drug by hemolytic experiments, which verified its non-hemolytic nature. These results imply that the enzymes have lots of potential for use in a range of industrial and medicinal applications.