Casitone-mediated expression of the prtP and prtM genes in Lactococcus lactis subsp. lactis BGIS29

Miladinov, Natasa; Kuipers, Oscar P.; Topisirović, Ljubiša

doi:10.1007/s00203-001-0361-7

Cited by 18 publications

(11 citation statements)

References 30 publications

(32 reference statements)

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“…lactis is similar to that of other LAB, including L. lactis (10,22,31), L. helveticus (14,39), L. delbrueckii subsp. bulgaricus (25), and L. rhamnosus (33).…”

Section: Resultsmentioning

confidence: 54%

“…The proteolytic system of lactococci is the best documented among LAB (10,18,22,23,28,29,31). The results of early experiments showed that in Lactococcus lactis, the synthesis of the cell wall proteinase PrtP during cell growth in peptide-rich medium is reduced compared to the rates of synthesis in milk or whey permeate medium with relatively low concentrations of peptides (18,28).…”

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confidence: 99%

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Characterization of the Pattern of α _s1 - and β-Casein Breakdown and Release of a Bioactive Peptide by a Cell Envelope Proteinase from Lactobacillus delbrueckii subsp. lactis CRL 581

Hébert

Mamone²,

Picariello³

et al. 2008

Appl Environ Microbiol

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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. The maximum 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, prolylleucine, leucylglycylglycine, or leucylproline, to the growth medium negatively affected CEP activity, whereas dipeptides without branched-chain amino acids had no effect on the enzyme's production. The carbon source and osmolites did not affect CEP activity. The CEP of L. delbrueckii subsp. lactis CRL 581 exhibited a mixed-type CEP I/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 ␣ s1 -and ␤-casein hydrolysates, respectively. By characterizing the peptide sequence in these hydrolysates, a pattern of ␣ s1 -and ␤-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 (antihypertensive and phosphopeptides) which are encrypted within the precursor protein could be visualized.

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“…lactis is similar to that of other LAB, including L. lactis (10,22,31), L. helveticus (14,39), L. delbrueckii subsp. bulgaricus (25), and L. rhamnosus (33).…”

Section: Resultsmentioning

confidence: 54%

mentioning

confidence: 99%

Characterization of the Pattern of α _s1 - and β-Casein Breakdown and Release of a Bioactive Peptide by a Cell Envelope Proteinase from Lactobacillus delbrueckii subsp. lactis CRL 581

Hébert

Mamone²,

Picariello³

et al. 2008

Appl Environ Microbiol

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“…A long distance between the transcription start and the start of the gene is characteristic of the proteinase genes of LAB (15,21,42). A minimal prtR promoter region determined by deletion analysis of (16,20,33,35). Therefore, we assume that when nutrition-dependent negative regulation is lifted, expression is up-regulated and the proteolytic activity of PrtR is elevated.…”

Section: Discussionmentioning

confidence: 99%

“…It has been shown that expression of prtP is repressed in the presence of rich nitrogen sources such as casein hydrolysates (Casitone), Casamino Acids, and specific dipeptides, e.g., leucylproline and prolylleucine (16,17,33,35). Regulation of the proteolytic system in lactobacilli is poorly studied, but it seems to be similar to that of L. lactis (20).…”

mentioning

confidence: 99%

Identification and Genetic Characterization of a Novel Proteinase, PrtR, from the Human Isolate Lactobacillus rhamnosus BGT10

Pastar¹,

Tonic²,

Golić³

et al. 2003

Appl Environ Microbiol

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A novel proteinase, PrtR, produced by the human vaginal isolate Lactobacillus rhamnosus strain BGT10 was identified and genetically characterized. The prtR gene and flanking regions were cloned and sequenced. The deduced amino acid sequence of PrtR shares characteristics that are common for other cell envelope proteinases (CEPs) characterized to date, but in contrast to the other cell surface subtilisin-like serine proteinases, it has a smaller and somewhat different B domain and lacks the helix domain, and the anchor domain has a rare sorting signal sequence. Furthermore, PrtR lacks the insert domain, which otherwise is situated inside the catalytic serine protease domain of all CEPs, and has a different cell wall spacer (W) domain similar to that of the cell surface antigen I and II polypeptides expressed by oral and vaginal streptococci. Moreover, the PrtR W domain exhibits significant sequence homology to the consensus sequence that has been shown to be the hallmark of human intestinal mucin protein. According to its ␣ S1 -and ␤-casein cleavage efficacy, PrtR is an efficient proteinase at pH 6.5 and is distributed throughout all L. rhamnosus strains tested. Proteinase extracts of the BGT10 strain obtained with Ca 2؉ -free buffer at pH 6.5 were proteolytically active. The prtR promoter-like sequence was determined, and the minimal promoter region was defined by use of prtR-gusA operon fusions. The prtR expression is Casitone dependent, emphasizing that nitrogen depletion elevates its transcription. This is in correlation with the catalytic activity of the PrtR proteinase.Lactic acid bacteria (LAB) have multiple amino acid auxotrophies, and in order to grow in protein-rich media they depend on the expression of a complex proteolytic system. Most of the genetic and biochemical studies of the LAB proteolytic system have focused on strains used in production of fermented milk products, and these data have been extensively reviewed (28,31,34). Essentially, a cell wall-bound extracellular proteinase (CEP) is responsible for the breakdown of casein, the major milk protein, into oligopeptides. Oligopeptides are then transported via oligopeptide transport systems into the bacterial cell, where the intracellular peptidases hydrolyze different oligopeptides to free amino acids.Three distinctly different genes encoding CEPs, referred to as prtP, prtB, and prtH (50), have been cloned and sequenced from dairy LAB. The prtP genes from a number of different Lactococcus lactis strains and from Lactobacillus paracasei NCDO151 have been cloned and sequenced (31). The different prtP genes encode proteinases with greater than 95% sequence identity. Distinct prt genes have been found in thermophilic lactobacilli. Genes encoding PrtB and PrtH from Lactobacillus delbrueckii subsp. bulgaricus NCDO1489 and Lactobacillus helveticus CNRZ32, respectively, have been sequenced and characterized (15,42). In order to produce the enzymatically active PrtP, L. lactis and L. paracasei require the presence of an upstream-located and divergently ...

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“…paracasei two genes are generally considered to be responsible for the initial breakdown of casein by dairy LAB: the prtP, and divergently oriented prtM gene Haandrikman et al 1989;Kojic et al 1991;Miladinov et al 2001;Pastar et al 2007).…”

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confidence: 99%

Variation in specificity of the PrtP extracellular proteinases in Lactococcus lactis and Lactobacillus paracasei subsp. paracasei

et al. 2009

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Comparison of cell-wall-bound extracellular proteinases (CEPs) from Lactobacillus paracasei (LBP) ssp. paracasei natural isolates BGHN14, BGAR75 and BGAR76 with Lactococcus lactis (LCL) ssp. cremoris Wg2, in their action on alpha(S1)-, beta- and kappa-casein was done. The CEPs of LBP strains were able to degrade alpha(S1)- and beta-caseins and their caseinolytic specificity depended on the type of buffer used. These CEPs, compared with LCL Wg2, differ in four amino acid residues in small segments predicted to be involved in substrate binding. The most striking features of this comparison are the presence of Ala instead of Ser(329) and the presence of Thr instead of Asn(256) and Ala(299), in the subtilisin-like region of the CEP in LBP natural isolates. Additional conservative amino acid substitution Leu to Ile(364) was found.

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Casitone-mediated expression of the prtP and prtM genes in Lactococcus lactis subsp. lactis BGIS29

Cited by 18 publications

References 30 publications

Characterization of the Pattern of α _s1 - and β-Casein Breakdown and Release of a Bioactive Peptide by a Cell Envelope Proteinase from Lactobacillus delbrueckii subsp. lactis CRL 581

Characterization of the Pattern of α _s1 - and β-Casein Breakdown and Release of a Bioactive Peptide by a Cell Envelope Proteinase from Lactobacillus delbrueckii subsp. lactis CRL 581

Identification and Genetic Characterization of a Novel Proteinase, PrtR, from the Human Isolate Lactobacillus rhamnosus BGT10

Variation in specificity of the PrtP extracellular proteinases in Lactococcus lactis and Lactobacillus paracasei subsp. paracasei

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Casitone-mediated expression of the prtP and prtM genes in Lactococcus lactis subsp. lactis BGIS29

Cited by 18 publications

References 30 publications

Characterization of the Pattern of α s1 - and β-Casein Breakdown and Release of a Bioactive Peptide by a Cell Envelope Proteinase from Lactobacillus delbrueckii subsp. lactis CRL 581

Characterization of the Pattern of α s1 - and β-Casein Breakdown and Release of a Bioactive Peptide by a Cell Envelope Proteinase from Lactobacillus delbrueckii subsp. lactis CRL 581

Identification and Genetic Characterization of a Novel Proteinase, PrtR, from the Human Isolate Lactobacillus rhamnosus BGT10

Variation in specificity of the PrtP extracellular proteinases in Lactococcus lactis and Lactobacillus paracasei subsp. paracasei

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Characterization of the Pattern of α _s1 - and β-Casein Breakdown and Release of a Bioactive Peptide by a Cell Envelope Proteinase from Lactobacillus delbrueckii subsp. lactis CRL 581

Characterization of the Pattern of α _s1 - and β-Casein Breakdown and Release of a Bioactive Peptide by a Cell Envelope Proteinase from Lactobacillus delbrueckii subsp. lactis CRL 581