Submit Manuscript  

Article Details


Refolding and Activation from Bacterial Inclusion Bodies of Trypsin I from Sardine (Sardinops sagax caerulea)

Author(s):

Manuel I. Carretas-Valdez , Francisco J. Cinco-Moroyoqui , Marina J. Ezquerra-Brauer, Enrique Marquez-Rios, Idania E. Quintero-Reyes, Alonso A. Lopez-Zavala and Aldo Alejandro Arvizu-Flores*   Pages 1 - 6 ( 6 )

Abstract:


Background: Trypsin from fish species is considered as a cold-adapted enzyme that may find potential biotechnological applications. In this work, the recombinant expression, refolding and activation of Trypsin I (TryI) from Monterey sardine (Sardinops sagax caerulea) are reported. Method: TryI was overexpressed in Escherichia coli BL21 as a fusion protein of trypsinogen with thioredoxin. Refolding of trypsinogen I was achieved by dialysis of bacterial inclusion bodies with a recovery of 16.32 mg per liter of Luria broth medium. Results: Before activation, the trypsinogen fusion protein did not show trypsin activity. Trypsinogen I was activated by adding 0.002 U of native TryI purified from the sardine pyloric caeca (nonrecombinant). The activated recombinant trypsin showed three times more activity than the nonrecombinant trypsin alone. Conclusion: The described protocol allowed obtaining sufficient amounts of recombinant TryI from Monterey sardine fish for further biochemical and biophysical characterization of its coldadaptation parameters.

Keywords:

Trypsin, refolding, sardine, cold-adapted, zymogen activation

Affiliation:

University of Sonora - Departamento de Investigacion y Posgrado en Alimentos Hermosillo, University of Sonora - Departamento de Investigacion y Posgrado en Alimentos Hermosillo, University of Sonora - Departamento de Investigacion y Posgrado en Alimentos Hermosillo, University of Sonora - Departamento de Investigacion y Posgrado en Alimentos Hermosillo, University of Sonora - Departamento de Ciencias de la Salud Cd. Obregon, University of Sonora - Departamento de Ciencias Quimico Biologicas Hermosillo, University of Sonora - Departamento de Ciencias Quimico Biologicas Hermosillo



Read Full-Text article