Purification and characterization of trypsin from Luphiosilurus alexandri pyloric cecum.

ABSTRACT: Trypsin from L. alexandri 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^-1, respectively. The purified enzyme was also as active as casein, as analyzed by zymograp

SUBMITTER: Dos Santos CWV 

PROVIDER: S-EPMC5613698 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature