TY - JOUR
T1 - Thermodynamic activation and structural analysis of trypsin i from Monterey sardine (Sardinops sagax caerulea)
AU - Arvizu-Flores, Aldo A.
AU - Quintero-Reyes, Idania E.
AU - Felix-Lopez, Martha
AU - Islas-Osuna, Maria A.
AU - Yepiz-Plascencia, Gloria
AU - Pacheco-Aguilar, Ramón
AU - Navare, Arti
AU - Fernández, Facundo M.
AU - Velazquez-Contreras, Enrique F.
AU - Sotelo-Mundo, Rogerio R.
AU - Castillo-Yañez, Francisco J.
N1 - 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.
PY - 2012/8/1
Y1 - 2012/8/1
N2 - 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.
AB - 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.
KW - Cold-adapted
KW - Molecular modelling
KW - Monterey sardine
KW - Sardinops sagax caerulea
KW - Thermodynamic activation parameters
KW - Trypsin
KW - cDNA
UR - http://www.scopus.com/inward/record.url?scp=84858335013&partnerID=8YFLogxK
U2 - 10.1016/j.foodchem.2012.01.111
DO - 10.1016/j.foodchem.2012.01.111
M3 - Artículo
SN - 0308-8146
VL - 133
SP - 898
EP - 904
JO - Food Chemistry
JF - Food Chemistry
IS - 3
ER -