CENTRO DE REFERENCIA PARA LACTOBACILOS
Unidad Ejecutora - UE
Characterization of the pattern of alpha s1- and beta-caseins breakdown and release of a bioactive peptide by a cell-envelope proteinase from Lactobacillus delbrueckii subsp. lactis CRL 581
HÉBERT EM.,; MAMONE G; PICARIELLO G; RAYA R; SAVOY DE GIORI G.; FERRANTI P; ADDEO F
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
AMER SOC MICROBIOLOGY
Año: 2008 vol. 74 p. 3682 - 3682
The cell envelope-associated proteinases (CEPs) of the lactobacilli have key roles in bacterial nutrition and contribute to the development of the organoleptic properties of fermented milk products as well, as they can release bioactive health-beneficial peptides from milk proteins. The influence of the peptide supply, carbohydrate source, and osmolites on the CEP activity of the cheese starter Lactobacillus delbrueckii subsp. lactis CRL 581 was investigated. The CEP activity levels were controlled by the peptide content of the growth medium. Themaximum activity was observed in a basal minimal defined medium, whereas in the presence of Casitone, Casamino Acids, or yeast extract, the synthesis of CEP was inhibited 99-, 70-, and 68-fold, respectively. The addition of specific di- or tripeptides containing branched-chain amino acids, such as leucylleucine, prolyl-leucine, leucylglycylglycine, or leucylproline, to the growth medium negatively af fected CEP activity, whereas dipeptides without branched-chain amino acids had no ef fect on the enzymes production. The carbon source and osmolites did not af fect CEP activity. The CEP of L. delbrueckii subsp. lactis CRL 581 exhibited a mixed-type CEPI/III variant caseinolytic specificity. Mass-spectrometric screening of the main peptide peaks isolated by reverse-phase high-pressure liquid chromatography allowed the identification of 33 and 32 peptides in the alpha s1- and beta-casein hydrolysates, respectively. By characterizing the peptide sequence in these hydrolysates, a pattern of alpha s1- and beta-casein breakdown was defined and is reported herein, this being the first report for a CEP of L. delbrueckii subsp. lactis. In this pattern, a series of potentially bioactive peptides (antihyper-tensive and phosphopeptides) which are encrypted within the precursor protein could be visualized.