TRYPSIN FROM JUMBO SQUID (Dosidicus gigas) HEPATOPANCREAS: PURIFICATION AND CHARACTERIZATION
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 elec...
| Autores: | , , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2017 |
| País: | México |
| Institución: | Universidad de Sonora |
| Repositorio: | Redalyc-USON |
| OAI Identifier: | oai:redalyc.org:33950011005 |
| Acceso en línea: | https://www.redalyc.org/articulo.oa?id=33950011005 https://www.redalyc.org/journal/339/33950011005/ https://www.redalyc.org/journal/339/33950011005/html/ https://www.redalyc.org/journal/339/33950011005/33950011005.epub https://www.redalyc.org/journal/339/33950011005/movil |
| Access Level: | acceso abierto |
| Palabra clave: | Multidisciplinarias (Ciencias Sociales) Squid Trypsin Hepatopancreas Dosidicus gigas Enzyme Purification |
| Sumario: | 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. |
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