Purification and characterization of a protease from the visceral mass of Mytella charruana and its evaluation to obtain antimicrobial peptides.

Journal: Food chemistry
PMID:

Abstract

This work describes purification of a protease from the visceral mass of the mussel Mytella charruana as well as evaluation of its ability to hydrolyze milk casein to generate antimicrobial peptides. The enzyme showed pI of 4.1 and a single polypeptide band of 83.1 kDa after SDS-PAGE. Sequence similarities with tropomyosin and myosin from mollusks were detected. The protease showed a trypsin-like activity with optimal temperature of 40 °C and stability in a wide pH range (3.0-9.0). K was 4.28 ± 0.34 mM of the synthetic substrate N-benzoyl-dl-arginyl-ρ-nitroanilide, whereas V was 0.056 ± 0.001 nmol min. The enzyme hydrolyzed casein, and the hydrolysate inhibited the growth of Escherichia coli, Micrococcus luteus, Bacillus subtilis, and Klebsiella pneumoniae at a minimal inhibitory concentration of 5.0 µg mL. In conclusion, the visceral mass of M. charruana contains a trypsin-like protease that can generate peptides from casein that have a bacteriostatic effect.

Authors

  • Leonardo Prezzi Dornelles
    Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil.
  • Maria de Fátima Deodato de Souza
    Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil.
  • Pollyanna Michelle da Silva
    Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil.
  • Thamara Figueiredo Procópio
    Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil.
  • Ricardo Salas Roldan Filho
    Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil.
  • Thâmarah de Albuquerque Lima
    Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil.
  • Ana Patrícia Silva de Oliveira
    Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil.
  • Russolina Benedeta Zingali
    Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
  • Patrícia Maria Guedes Paiva
    Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil.
  • Emmanuel Viana Pontual
    Departamento de Morfologia e Fisiologia Animal, Universidade Federal Rural de Pernambuco, Recife, Brazil.
  • Thiago Henrique Napoleão
    Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil. Electronic address: thiago.napoleao@ufpe.br.