TY - JOUR
T1 - Refolding and activation from bacterial inclusion bodies of trypsin i from sardine (sardinops sagax caerulea)
AU - Carretas-Valdez, Manuel I.
AU - Cinco-Moroyoqui, Francisco J.
AU - Ezquerra-Brauer, Marina J.
AU - Marquez-Rios, Enrique
AU - Quintero-Reyes, Idania E.
AU - Lopez-Zavala, Alonso A.
AU - Arvizu-Flores, Aldo A.
PY - 2019/1/1
Y1 - 2019/1/1
N2 - © 2019 Bentham Science Publishers. 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.
AB - © 2019 Bentham Science Publishers. 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.
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U2 - 10.2174/0929866525666181019161114
DO - 10.2174/0929866525666181019161114
M3 - Article
C2 - 30338728
SP - 170
EP - 175
JO - Protein and Peptide Letters
JF - Protein and Peptide Letters
SN - 0929-8665
ER -