Characterization of the trypsin-III from Monterey sardine (Sardinops caeruleus): Insights on the cold-adaptation from the A236N mutant

Manuel I. Carretas-Valdez, Elena N. Moreno-Cordova, Brisa G. Ibarra-Hernandez, Francisco J. Cinco-Moroyoqui, Francisco J. Castillo-Yañez, Sergio Casas-Flores, Pablo S. Osuna-Amarillas, Maria A. Islas-Osuna*, Aldo A. Arvizu-Flores

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Trypsins (E.C. 3.4.21.4) are digestive enzymes that catalyze the hydrolysis of peptide bonds containing arginine and lysine residues. Some trypsins from fish species are active at temperatures just above freezing, and for that are called cold-adapted enzymes, having many biotechnological applications. In this work, we characterized a recombinant trypsin-III from Monterey sardine (Sardinops caeruleus) and studied the role of a single residue on its cold-adapted features. The A236N mutant from sardine trypsin-III showed higher activation energy for the enzyme-catalyzed reaction, it was more active at higher temperatures, and exhibited a higher thermal stability than the wild-type enzyme, suggesting a key role of this residue. The thermodynamic activation parameters revealed an increase in the activation enthalpy for the A236N mutant, suggesting the existence of more intramolecular contacts during the activation step. Molecular models for both enzymes suggest that a hydrogen-bond involving N236 may contact the C-terminal α-helix to the vicinity of the active site, thus affecting the biochemical and thermodynamic properties of the enzyme.

Original languageEnglish
Pages (from-to)2701-2710
Number of pages10
JournalInternational Journal of Biological Macromolecules
Volume164
DOIs
StatePublished - 1 Dec 2020

Bibliographical note

Publisher Copyright:
© 2020 Elsevier B.V.

Keywords

  • Activation energy
  • Cold-adapted
  • ITC kinetics
  • Monterey sardine
  • Trypsin-III

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