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2017
- handle: 10670/1.z81vfb
- 0378-1844
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http://www.redalyc.org/revista.oa
Interciencia
Mots-clés
Multidisciplinarias (Ciencias Sociales) Squid Trypsin Hepatopancreas Dosidicus gigas Enzyme PurificationSujets proches
Trypsase Alkali proteinaseCiter ce document
Ana Gloria Villalba-Villalba et al., « TRYPSIN FROM JUMBO SQUID (Dosidicus gigas) HEPATOPANCREAS: PURIFICATION AND CHARACTERIZATION », Interciencia, ID : 10670/1.z81vfb
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Résumé
"Trypsin was purified from the hepatopancreas of Dosidicus gigas by f ractionation with ammonium sulfate (30-70% saturation), gel filtration, affinity and ion exchange chromatography. The molecular weight of the trypsin obtained was ~23.5kDa according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and, showed a single band in the zymogram. The enzyme exhibited maximal activity at pH 8.5 and 40ºC, using BAPNA as substrate. It was effectively inhibited by phenyl methyl sulfonyl fluoride (PMSF) (98%), and Nα-p-Tosyl-L-lysine chloromethyl ketone (TLCK) (99%). Enzyme activity was affected by the following ions in decreasing order: Hg2+, Fe2+, Cu2+, Li1+, Mg2+, K1+, Mn2+, Ca2+. Trypsin activity decreased continuously as NaCl concentration increased from 0 to 30%. Km and kcat values were 0.085 ±1.45mM and 1.76 ±0.12s-1, respectively. The results suggest that the purified enzyme a potential agent to be used in biotechnological processes."