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

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

  • Int J Biol Macromol. 2020 Dec 1;164:2701-2710. doi: 10.1016/j.ijbiomac.2020.08.136.
Manuel I Carretas-Valdez 1,  Elena N Moreno-Cordova 2,  Brisa G Ibarra-Hernandez 1,  Francisco J Cinco-Moroyoqui 1,  Francisco J Castillo-Yañez 2,  Sergio Casas-Flores 3,  Pablo S Osuna-Amarillas 4,  Maria A Islas-Osuna 5,  Aldo A Arvizu-Flores 6
Affiliations

Affiliations

  • 1 Universidad de Sonora, Departamento de Investigación y Posgrado en Alimentos, Blvd. Luis Encinas y Blvd. Rosales s/n, Hermosillo, Sonora 83000, Mexico.
  • 2 Universidad de Sonora, Departamento de Ciencias Químico-Biológicas, Blvd. Luis Encinas y Blvd. Rosales s/n, Hermosillo, Sonora 83000, Mexico.
  • 3 IPICYT, División de Biología Molecular, Camino a la Presa San José 2055, Col. Lomas 4a sección, San Luis Potosí, San Luis Potosí 78216, Mexico.
  • 4 Universidad Estatal de Sonora, Carretera Navojoa-Huatabampo km 5, Navojoa, Sonora 85874, Mexico.
  • 5 Centro de Investigación en Alimentación y Desarrollo, Laboratorio de Genética y Biología Molecular de Plantas, Carr. Gustavo Enrique Astiazarán Rosas, N0. 46. Col. La Victoria, Hermosillo, Sonora 83304, Mexico. Electronic address: islasosu@ciad.mx.
  • 6 Universidad de Sonora, Departamento de Ciencias Químico-Biológicas, Blvd. Luis Encinas y Blvd. Rosales s/n, Hermosillo, Sonora 83000, Mexico. Electronic address: aldo.arvizu@unison.mx.
PMID: 32827617 DOI: 10.1016/j.ijbiomac.2020.08.136
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.

Keywords

Activation energy; Cold-adapted; ITC kinetics; Monterey sardine; Trypsin-III.

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