caa a22 4500 605502110 CHVBK 20210128100604.0 cr unu---uuuuu 210128e20150501xx s 000 0 eng 10.1007/s00253-014-6258-6 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6258-6 Characterization of the mature cell surface proteinase of Lactobacillus delbrueckii subsp. lactis CRL 581 [Elektronische Daten] [Josefina Villegas, Lucía Brown, Graciela Savoy de Giori, Elvira Hebert] The cell envelope-associated proteinase (CEP) of Lactobacillus delbrueckii subsp. lactis CRL 581 (PrtL) has an essential role in bacterial growth, contributes to the flavor and texture development of fermented products, and can release bioactive health-beneficial peptides during milk fermentation. The genome of L. delbrueckii subsp. lactis CRL 581 possesses only one gene that encodes PrtL, which consists of 1924 amino acids and is a multidomain protein anchored to the cell via its W domain. PrtL was extracted from the cell under high ionic strength conditions using NaCl, suggesting an electrostatic interaction between the proteinase and the cell envelope. The released PrtL was purified and biochemically characterized; its activity was maximal at temperatures between 37 and 40°C and at pH between 7 and 8. Under optimal conditions, PrtL exhibited higher affinity for succinyl-alanyl-alanyl-prolyl-phenylalanine-p-nitroanilide than for succinyl-alanyl-glutamyl-prolyl-phenylalanine-p-nitroanilide, while methoxy-succinyl-arginyl-prolyl-tyrosyl-p-nitroanilide was not degraded. A similar α- and β-casein degradation pattern was observed with the purified and the cell envelope-bound proteinase. Finally, on the basis of its specificity towards caseins and the unique combination of amino acids at residues thought to be involved in substrate specificity, PrtL can be classified as a representative of a new group of CEP. Springer-Verlag Berlin Heidelberg, 2014 Lactic acid bacteria nationallicence Proteinase nationallicence Casein nationallicence Proteolytic activity nationallicence Caseinolytic specificity nationallicence Villegas Josefina Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, 4000, San Miguel de Tucumán, Argentina aut Brown Lucía Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, 4000, San Miguel de Tucumán, Argentina aut Savoy de Giori Graciela Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, 4000, San Miguel de Tucumán, Argentina aut Hebert Elvira Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, 4000, San Miguel de Tucumán, Argentina aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/10(2015-05-01), 4277-4286 0175-7598 99:10<4277 2015 99 253 https://doi.org/10.1007/s00253-014-6258-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6258-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Villegas Josefina Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, 4000, San Miguel de Tucumán, Argentina aut NATIONALLICENCE 700 1- Brown Lucía Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, 4000, San Miguel de Tucumán, Argentina aut NATIONALLICENCE 700 1- Savoy de Giori Graciela Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, 4000, San Miguel de Tucumán, Argentina aut NATIONALLICENCE 700 1- Hebert Elvira Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, 4000, San Miguel de Tucumán, Argentina aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/10(2015-05-01), 4277-4286 0175-7598 99:10<4277 2015 99 253