Metal-sensitive and thermostable trypsin from the crevalle jack (Caranx hippos) pyloric caeca: purification and characterization
Open Access
- 10 October 2013
- journal article
- Published by Springer Science and Business Media LLC in BMC Chemistry
- Vol. 7 (1), 166
- https://doi.org/10.1186/1752-153x-7-166
Abstract
Background: Over the past decades, the economic development and world population growth has led to increased for food demand. Increasing the fish production is considered one of the alternatives to meet the increased food demand, but the processing of fish leads to by-products such as skin, bones and viscera, a source of environmental contamination. Fish viscera have been reported as an important source of digestive proteases with interesting characteristics for biotechnological processes. Thus, the aim of this study was to purify and to characterize a trypsin from the processing by-products of crevalle jack (Caranx hippos) fish. Results: A 27.5 kDa trypsin with N-terminal amino acid sequence IVGGFECTPHVFAYQ was easily purified from the pyloric caeca of the crevalle jack. Its physicochemical and kinetic properties were evaluated using N-α-benzoyl-DL-arginine-p-nitroanilide (BApNA) as substrate. In addition, the effects of various metal ions and specific protease inhibitors on trypsin activity were determined. Optimum pH and temperature were 8.0 and 50°C, respectively. After incubation at 50°C for 30 min the enzyme lost only 20% of its activity. Km, kcat, and kcat/Km values using BApNA as substrate were 0.689 mM, 6.9 s-1, and 10 s-1 mM-1, respectively. High inhibition of trypsin activity was observed after incubation with Cd2+, Al3+, Zn2+, Cu2+, Pb2+, and Hg2+ at 1 mM, revealing high sensitivity of the enzyme to metal ions. Conclusions: Extraction of a thermostable trypsin from by-products of the fishery industry confirms the potential of these materials as an alternative source of these biomolecules. Furthermore, the results suggest that this trypsin-like enzyme presents interesting biotechnological properties for industrial applications.Keywords
This publication has 36 references indexed in Scilit:
- Giant Amazonian fish pirarucu (Arapaima gigas): Its viscera as a source of thermostable trypsinFood Chemistry, 2012
- Trypsin from zebra blenny (Salaria basilisca) viscera: Purification, characterisation and potential application as a detergent additiveFood Chemistry, 2012
- Purification and partial characterisation of a trypsin from the processing waste of the silver mojarra (Diapterus rhombeus)Food Chemistry, 2011
- Purification, characterization and substrate specificity of a trypsin from the Amazonian fish tambaqui (Colossoma macropomum)Biochemical and Biophysical Research Communications, 2010
- Alkaline proteinase from intestine of Nile tilapia (Oreochromis niloticus)Process Biochemistry, 2005
- Studies on production of thermostable alkaline protease from thermophilic and alkaliphilic Bacillus sp. JB-99 in a chemically defined mediumProcess Biochemistry, 2001
- PARTIAL PURIFICATION AND CHARACTERIZATION OF A THERMOSTABLE TRYPSIN FROM PYLORIC CAECA OF TAMBAQUI (COLOSSOMA MACROPOMUM)Journal of Food Biochemistry, 2001
- Trypsin and trypsinogen from an Antarctic fish: molecular basis of cold adaptationBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1996
- The structure of bovine trypsin : Electron density maps of the inhibited enzyme at 5 Å and at 2·7 Å resolutionJournal of Molecular Biology, 1974
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970