title: Protease and esterase activity of staphylococci.
abstract: The aim of this work was to characterize protease and esterase activities of staphylococci in order to establish if they could contribute to the release of amino acids and short-chain fatty acids during ripening of fermented sausages. Eighteen Staphylococcus strains belonging to the species Staphylococcus xylosus (5), S. saprophyticus (3), S. equorum (4), S. carnosus (4) and S. simulans (2), previously isolated from different types of Southern Italian fermented sausages, were screened for proteinase, aminopeptidase and esterase activities. Most of the staphylococci strains lacked detectable levels of proteinase activity against casein-fluorescein isothiocynate. In the active strains, this activity was extracellular or cell-envelope associated. The studied staphylococci strains also showed low levels of aminopeptidase activities, which preferentially hydrolysed substrates containing L-methionine, L-leucine and L-phenylalanine as N-terminal residue. In contrast, all staphylococcal strains possessed significant activity against short-chain fatty acid esters. The maximum esterase activities were detected in whole-cell suspensions and cell-free extracts and to a lesser extent in the extracellular medium. The substrates preferentially hydrolysed were rho-nitrophenyl (rho-NP) butyrate and rho-NP caprylate and, secondly, rho-NP palmitate. The extracellular extracts of most of the strains were only active against rho-NP butyrate except for those of S. equorum (SI3, SI4) and S. simulans (Ssm12, Ssm21), which also hydrolysed rho-NP caprylate and rho-NP palmitate. The cell-free extracts and whole cells were mainly active against rho-NP butyrate and rho-NP caprylate, showing activity levels from 1760 U/mg of proteins to 54 U/mg of proteins and from 12,200 U/mg of proteins to 133 U/mg of proteins respectively. These activities were especially high in the strains that belonged to S. xylosus and S. equorum species. The diversity of the studied metabolic properties and, especially, the esterase activities in different staphylococcal species and even strains of the same species emphasize the relevance of these in vitro characterization studies for a rational selection of new starter cultures.
authors.affiliation: Dipartimento di Scienza degli Alimenti Universitá degli Studi di Napoli Federico II, 80055 Portici, Naples, Italy. acasabur@unina.it
authors.forename: Annalisa
authors.initials: A
authors.lastname: Casaburi
authors.forename: Francesco
authors.initials: F
authors.lastname: Villani
authors.forename: Fidel
authors.initials: F
authors.lastname: Toldrá
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Sanz
mesh_terms: D000626:Aminopeptidases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D004950:Esterases / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D005511:Food Handling / Q000379:methods*; D005516:Food Microbiology; D006868:Hydrolysis; D008461:Meat Products / Q000382:microbiology*; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D013045:Species Specificity; D013210:Staphylococcus / Q000145:classification* / Q000201:enzymology*; D013379:Substrate Specificity; D013552:Swine
chemical_list: D004950:Esterases; D010447:Peptide Hydrolases; D000626:Aminopeptidases