Giant Amazonian fish pirarucu (Arapaima gigas): Its viscera as a source of thermostable trypsin

• Autor(a) principal: Freitas-Junior, Augusto C. V.
• Data de Publicação: 2012
• Outros Autores: Costa, Helane M. S., Icimoto, Marcelo Yudi [UNIFESP], Hirata, Izaura Yoshico [UNIFESP], Marcondes, Marcelo [UNIFESP], Carvalho, Luiz B., Oliveira, Vitor [UNIFESP], Bezerra, Ranilson S.
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UNIFESP
• Texto Completo: http://dx.doi.org/10.1016/j.foodchem.2012.02.056; http://repositorio.unifesp.br/handle/11600/35176
• topic: Arapaima gigas; Air-breathing fish; Fish processing waste; Digestive enzymes; Proteases; Trypsin purification

A trypsin was purified from pyloric caeca of pirarucu (Arapaima gigas). the effect of metal ions and protease inhibitors on its activity and its physicochemical and kinetic properties, as well its N-terminal sequence, were determined. A single band (28.0 kDa) was observed by SDS-PAGE. Optimum pH and temperature were 9.0 and 65 degrees C, respectively. the enzyme was stable after incubation for 30 min in a wide pH range (6.0-11.5) and at 55 degrees C. the kinetic parameters K-m, k(cat) and k(cat)/K-m were 0.47 +/- 0.042 mM, 1.33 s(-1) and 2.82 s(-1) mM(-1), respectively, using BApNA as substrate. This activity was shown to be very sensitive to some metal ions, such as Fe2+, Hg2+, Zn2+, Al3+, Pb2+, and was highly inhibited by trypsin inhibitors. the trypsin N-terminal sequence IVGGYECPRNSVPYQ was found. the features of this alkaline peptidase suggest that it may have potential for industrial applications (e.g. food and detergent industries). (C) 2012 Elsevier B.V. All rights reserved.