Purification and characterization of trypsin from Luphiosilurus alexandri pyloric cecum

• Published in: Biochemistry and biophysics reports Vol. 8; pp. 29 - 33
• Main Authors: Dos Santos, Claudio Wilian Victor, da Costa Marques, Maria Elizabeth, de Araújo Tenório, Humberto, de Miranda, Edma Carvalho, Vieira Pereira, Hugo Juarez
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier 01.12.2016
• ISSN: 2405-5808; 2405-5808
• DOI: 10.1016/j.bbrep.2016.08.003

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.