Purification, characterization and substrate specificity of a trypsin from the Amazonian fish tambaqui (Colossoma macropomum)

Purification, characterization and substrate specificity of a trypsin from the Amazonian fish tambaqui (Colossoma macropomum)

Author Marcuschi, Marina Google Scholar
Esposito, Talita S. Google Scholar
Machado, Mauricio F. M. Autor UNIFESP Google Scholar
Hirata, Izaura Y. Autor UNIFESP Google Scholar
Machado, Marcelo F. M. Autor UNIFESP Google Scholar
Silva, Marcia V. Google Scholar
Carvalho, Luiz B. Google Scholar
Oliveira, Vitor Autor UNIFESP Google Scholar
Bezerra, Ranilson S. Google Scholar
Institution Universidade Federal de Pernambuco (UFPE)
Universidade Federal de São Paulo (UNIFESP)
Abstract An enzyme was purified from the pyloric caecum of tambaqui (Colossoma macropomum) through heat treatment, ammonium sulfate fractionation, Sephadex (R) G-75 and p-aminobenzamidine-agarose affinity chromatography. the enzyme had a molecular mass of 23.9 kDa, NH(2)-terminal amino acid sequence of IVGGYECKAHSQPHVSLNI and substrate specificity for arginine at P1, efficiently hydrolizing substrates with leucine and lysine at P2 and serine and arginine at P1'. Using the substrate z-FR-MCA, the enzyme exhibited greatest activity at pH 9.0 and 50 degrees C, whereas, with BAPNA activity was higher in a pH range of 7.5-11.5 and at 70 degrees C. Moreover, the enzyme maintained ca. 60% of its activity after incubated for 3 h at 60 degrees C. the enzymatic activity significantly decreased in the presence of TLCK, benzamidine (trypsin inhibitors) and PMSF (serine protease inhibitor). This source of trypsin may be an attractive alternative for the detergent and food industry. (C) 2010 Elsevier Inc. All rights reserved.
Keywords FRET peptide
Processing waste
Specific cleavage site
Tropical fish
Trypsin
Language English
Sponsor Ministry of Fisheries and Aquaculture
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
FINEP
FACEPE
PETROBRAS
Date 2010-06-04
Published in Biochemical and Biophysical Research Communications. San Diego: Academic Press Inc Elsevier Science, v. 396, n. 3, p. 667-673, 2010.
ISSN 0006-291X (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 667-673
Origin http://dx.doi.org/10.1016/j.bbrc.2010.04.155
Access rights Closed access
Type Article
Web of Science ID WOS:000278710200016
URI http://repositorio.unifesp.br/handle/11600/32621

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