Thermodynamic activation and structural analysis of trypsin i from Monterey sardine (Sardinops sagax caerulea)

Aldo A. Arvizu-Flores, Idania E. Quintero-Reyes, Martha Felix-Lopez, Maria A. Islas-Osuna, Gloria Yepiz-Plascencia, Ramón Pacheco-Aguilar, Arti Navare, Facundo M. Fernández, Enrique F. Velazquez-Contreras, Rogerio R. Sotelo-Mundo*, Francisco J. Castillo-Yañez

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

In this work, we report the molecular characterisation of trypsin I (Try I) from Monterey sardine (Sardinops sagax caerulea). Aspects such as thermodynamic activation parameters, molecular model and cDNA-deduced amino acid sequence allow a more in depth understanding of its activity at low temperatures. The analysis of the thermodynamic activation parameters suggests that this molecule is a cold-adapted protease. From the molecular cloning, we deduced the amino acid sequence and predicted a theoretical structural model of sardine Try I with a classical trypsin fold. Cold-adaptation of this enzyme probably comes from amino acid replacement of key residues to improve flexibility at low temperature, thus increasing k cat. The cold-adaptation of sardine Try I opens a wide range of biotechnological applications for this protease and also it is interesting from the structure function relationship point of view of serine protease proteins.

Original languageEnglish
Pages (from-to)898-904
Number of pages7
JournalFood Chemistry
Volume133
Issue number3
DOIs
StatePublished - 1 Aug 2012

Bibliographical note

Funding Information:
We acknowledge support from CONACYT (Mexico´s National Council of Science and Technology) Grant No. 60477 to F.J. Castillo-Yañez, grants E0007-2011-01-179940 and CB-2009-01-131859 to R. Sotelo-Mundo for molecular modeling license and computer hardware and “Apoyos Complementarios para la Consolidacio´n Institucional de Grupos de Investigacio´n: Retencio´n 121269” to A.A. Arvizu-Flores. M. Félix-López and I.E. Quintero-Reyes thanks a graduate scholarship also from CONACYT-Mexico. M.A. Islas-Osuna thanks CONACYT for a sabbatical scholarship at University of Sonora. E.F. Velazquez-Contreras acknowledges support from Subsecretaría de Educación Superior/SEP grant P/PIFI 2001−26-F0−08 for equipment instrumentation. We thank Alma B. Peregrino-Uriarte, M.Sc., Karina Garcia-Orozco, M.Sc. and Rocio Sugich-Miranda, Ph.D. for help and advice with cDNA cloning, protein purification and thermodynamics. We also thank Felipe Isac-Martinez, Monica Resendiz-Sandoval, Cesar Otero-León and Gerardo Reyna-Cañez for technical support in all stages of this research.

Keywords

  • Cold-adapted
  • Molecular modelling
  • Monterey sardine
  • Sardinops sagax caerulea
  • Thermodynamic activation parameters
  • Trypsin
  • cDNA

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