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, Aldo A. Arvizu-Flores*

Journal Name: Protein & Peptide Letters

Volume 26 , Issue 3 , 2019

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Graphical 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.

Methods: 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, recombinant expression.

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Article Details

Year: 2019
Page: [170 - 175]
Pages: 6
DOI: 10.2174/0929866525666181019161114
Price: $65

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