Bioengineering and Environmental Centre, Indian Institute of Chemical Technology, Tarnaka, Hyderabad 500607, India.
Int J Biol Macromol. 2013 Oct;61:479-86. doi: 10.1016/j.ijbiomac.2013.07.009. Epub 2013 Jul 30.
This study shows the purification and characterization of metalloprotease (serralysin) with fibrin and fibrinogenolytic property, from the newly isolated Serratia marcescens RSPB11. This protein macro molecule was more stable over a wide range of pH (6-10) and the temperatures up to 60 °C. It showed optimum enzyme activity at pH 9.0 and at a temperature of 37 °C. Inhibitory analysis revealed that this enzyme is metalloprotease and its enzyme activity could be regained by the addition of Co(2+), Cu(2+), Fe(2+), Mg(2+)and Zn(2+) ions after chelation of ions with EDTA. This enzyme showed the Michaelis-Menten's constant Km (1.261 mg/ml) for its substrate, casein and the observed maximum attainable velocity was Vmax (24,842 U/min). The purified enzyme showed an apparent molecular mass of approximately 50 kDa in SDS-PAGE. The results also suggested that this serralysin is having potential application thrombolytic therapy.
本研究展示了一种新型分离的粘质沙雷氏菌 RSPB11 来源的具有纤维蛋白和纤维蛋白原水解特性的金属蛋白酶（糜蛋白酶）的纯化和特性。这种蛋白质大分子在较宽的 pH（6-10）和温度范围内（高达 60°C）更稳定。它在 pH 9.0 和 37°C 时表现出最佳的酶活性。抑制分析表明，该酶为金属蛋白酶，其酶活性在与 EDTA 螯合后，可通过添加 Co(2+)、Cu(2+)、Fe(2+)、Mg(2+)和 Zn(2+)离子而恢复。该酶对其底物酪蛋白的米氏常数 Km（1.261mg/ml），观察到的最大可达到的速度为 Vmax（24842U/min）。纯化的酶在 SDS-PAGE 中显示出约 50 kDa 的表观分子量。结果还表明，这种糜蛋白酶在溶栓治疗方面具有潜在的应用价值。
