从亚历山大卢非鱼幽门盲囊中纯化和鉴定胰蛋白酶。
Purification and characterization of trypsin from Luphiosilurus alexandri pyloric cecum.
作者信息
Dos Santos Claudio Wilian Victor, da Costa Marques Maria Elizabeth, de Araújo Tenório Humberto, de Miranda Edma Carvalho, Vieira Pereira Hugo Juarez
机构信息
Instituto de Química e Biotecnologia, Universidade Federal de Alagoas, Avenida Lourival Melo Mota, s/n, 57072-970 Maceió, AL, Brazil.
出版信息
Biochem Biophys Rep. 2016 Aug 11;8:29-33. doi: 10.1016/j.bbrep.2016.08.003. eCollection 2016 Dec.
Trypsin from . was purified using only two purification processes: ammonium sulfate precipitation and anion exchange liquid chromatography in DEAE-Sepharose. Trypsin mass was estimated as 24 kDa through SDS-PAGE, which showed only one band in silver staining. The purified enzyme showed an optimum temperature and pH of 50 °C and 9.0, respectively. Stability was well maintained, with high levels of activity at a pH of up to 11.0, including high stability at a temperature of up to 50 °C after 60 min of incubation. The inhibition test demonstrated strong inhibition by PMSF, a serine protease inhibitor, and Kinetic constants km and kcat for BAPNA were 0.517 mM and 5.0 S, respectively. The purified enzyme was also as active as casein, as analyzed by zymography. Therefore, we consider trypsin a promising enzyme for industrial processes, owing to its stability in a wide range of pH and temperature and activity even under immobilization.
来自……的胰蛋白酶仅通过两种纯化方法进行纯化：硫酸铵沉淀和在DEAE-琼脂糖中的阴离子交换液相色谱法。通过SDS-PAGE估计胰蛋白酶质量为24 kDa，银染显示只有一条带。纯化后的酶的最适温度和pH分别为50°C和9.0。稳定性良好，在pH高达11.0时具有高水平活性，包括在50°C孵育60分钟后仍具有高稳定性。抑制试验表明，丝氨酸蛋白酶抑制剂PMSF具有强烈抑制作用，BAPNA的动力学常数km和kcat分别为0.517 mM和5.0 S。经酶谱分析，纯化后的酶对酪蛋白也具有活性。因此，我们认为胰蛋白酶因其在广泛的pH和温度范围内具有稳定性且即使在固定化条件下也具有活性，是一种用于工业过程的有前景的酶。
Zotero 插件