Specific aminopeptidases of indigenous Lactobacillus brevis and Lactobacillus plantarum

  • Khadidja Belkheir
  • Salima Roudj
  • Halima Zadi Karam
  • Nour Eddine Karam
Keywords: Camel milk, Lactobacillus, proteolysis, leucyl aminopeptidase activity.

Abstract

Lactic acid bacteria play an important role in milk coagulation and cheese ripening. To select strains  showing interesting industrial features, two indigenous lactobacilli (Lactobacillus brevis and Lactobacillus plantarum) were studied for aminopeptidase activity. Cell and cells free extract were tested for leucyl aminopeptidase activity on the chromogenic leucyl-p-nitroanilide substrate. Intracellu-lar and membrane enzymes were solubilized with glycine /lyzozyme treatment then purified by ammonium sulphate precipitation followed by Sephadex G100 and diethylaminoethyl (DEAE) ions exchange chromatography’s separation. The molecular weight of denatured proteins was estimated on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE). Effects of several parameters, pH, temperature, some ions and inhibitors on purified enzyme activity were studied. Cellular amino-peptidase activity was higher for CHTD 27 strain than BH14 strain. No aminopeptidase activity was noted in the cell free extract. The results of chromatography sephadex G100 combined to those of electrophoresis allowed suggesting a dimer structure for the native enzyme. The Lb CHTD27 purified enzyme showed maximal activity at pH 6.6 and at 40°C. This enzyme was partially inhibited by ethylenediamine acetic acid (EDTA) and Cu<suP2+ions but increased by Na2+ and Co2+ ions. The aminopeptidase extracted from Lb BH14 was inhibited by EDTA and phenylmethanesulfonylfluoride or phenylmethylsulfonyl fluoride (PMSF), its maximal activity was observed at pH 7.5 and 40°C. In addition to other characteristics as proteolysis and autolysis, in this paper we showed that both studied strains were also able to degrade peptide with specific peptidases which are important characters in cheese manufacturing.

Key words: Camel milk, Lactobacillus, proteolysis, leucyl aminopeptidase activity.

Published
2016-01-27
Section
Articles

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eISSN: 1684-5315