Proteolytic activities, peptide utilization and oligopeptide transport systems of wild Lactococcus lactis strains

2010 ◽  
Vol 20 (3) ◽  
pp. 156-162 ◽  
Author(s):  
A. Picon ◽  
M.A. García-Casado ◽  
M. Nuñez
Keyword(s):  
Lactococcus Lactis ◽  
Transport Systems ◽  
Proteolytic Activities ◽  
Oligopeptide Transport

scholarly journals The Oligopeptide Transport System Is Essential for the Development of Natural Competence in Streptococcus thermophilus Strain LMD-9

2009 ◽  
Vol 191 (14) ◽  
pp. 4647-4655 ◽  
Author(s):  
Rozenn Gardan ◽  
Colette Besset ◽  
Alain Guillot ◽  
Christophe Gitton ◽  
Véronique Monnet
Keyword(s):  
Quorum Sensing ◽  
Streptococcus Thermophilus ◽  
Defined Medium ◽  
Mass Spectrometry Analysis ◽  
Transport Systems ◽  
Label Free ◽  
Spectrometry Analysis ◽  
Proteomic Approach ◽  
Liquid Chromatography Tandem Mass ◽  
Oligopeptide Transport

ABSTRACT In gram-positive bacteria, oligopeptide transport systems, called Opp or Ami, play a role in nutrition but are also involved in the internalization of signaling peptides that take part in the functioning of quorum-sensing pathways. Our objective was to reveal functions that are controlled by Ami via quorum-sensing mechanisms in Streptococcus thermophilus, a nonpathogenic bacterium widely used in dairy technology in association with other bacteria. Using a label-free proteomic approach combining one-dimensional electrophoresis with liquid chromatography-tandem mass spectrometry analysis, we compared the proteome of the S. thermophilus LMD-9 to that of a mutant deleted for the subunits C, D, and E of the ami operon. Both strains were grown in a chemically defined medium (CDM) without peptides. We focused our attention on proteins that were no more detected in the ami deletion mutant. In addition to the three subunits of the Ami transporter, 17 proteins fulfilled this criterion and, among them, 7 were similar to proteins that have been identified as essential for transformation in S. pneumoniae. These results led us to find a condition of growth, the early exponential state in CDM, that allows natural transformation in S. thermophilus LMD-9 to turn on spontaneously. Cells were not competent in M17 rich medium. Furthermore, we demonstrated that the Ami transporter controls the triggering of the competence state through the control of the transcription of comX, itself controlling the transcription of late competence genes. We also showed that one of the two oligopeptide-binding proteins of strain LMD-9 plays the predominant role in the control of competence.

scholarly journals Utilization of Oligopeptides by Listeria monocytogenes Scott A

1998 ◽  
Vol 64 (3) ◽  
pp. 1059-1065 ◽  
Author(s):  
Annette Verheul ◽  
Frank M. Rombouts ◽  
Tjakko Abee
Keyword(s):  
Listeria Monocytogenes ◽  
Transport System ◽  
Growth Stimulation ◽  
Sole Source ◽  
Fermented Milk ◽  
Defined Medium ◽  
Peptide Transport ◽  
Transport Systems ◽  
Oligopeptide Transport System ◽  
Oligopeptide Transport

ABSTRACT For effective utilization of peptides, Listeria monocytogenes possesses two different peptide transport systems. The first one is the previously described proton motive force (PMF)-driven di- and tripeptide transport system (A. Verheul, A. Hagting, M.-R. Amezaga, I. R. Booth, F. M. Rombouts, and T. Abee, Appl. Environ. Microbiol. 61:226–233, 1995). The present results reveal that L. monocytogenes possesses an oligopeptide transport system, presumably requiring ATP rather than the PMF as the driving force for translocation. Experiments to determine growth in a defined medium containing peptides of various lengths suggested that the oligopeptide permease transports peptides of up to 8 amino acid residues. Peptidase activities towards several oligopeptides were demonstrated in cell extract from L. monocytogenes, which indicates that upon internalization, the oligopeptides are hydrolyzed to serve as sources of amino acids for growth. The peptide transporters of the nonproteolytic L. monocytogenes might play an important role in foods that harbor indigenous proteinases and/or proteolytic microorganisms, since Pseudomonas fragi as well as Bacillus cereus was found to enhance the growth ofL. monocytogenes to a large extent in a medium in which the milk protein casein was the sole source of nitrogen. In addition, growth stimulation was elicited in this medium when casein was hydrolyzed by using purified protease from Bacillus licheniformis. The possible contribution of the oligopeptide transport system in the establishment of high numbers of L. monocytogenes cells in fermented milk products is discussed.

Identification and functional characterisation of cellobiose and lactose transport systems in Lactococcus lactis IL1403

2007 ◽  
Vol 189 (3) ◽  
pp. 187-196 ◽  
Author(s):  
Magdalena Kowalczyk ◽  
Muriel Cocaign-Bousquet ◽  
Pascal Loubiere ◽  
Jacek Bardowski
Keyword(s):  
Lactococcus Lactis ◽  
Transport Systems ◽  
Functional Characterisation ◽  
Lactis Il1403 ◽  
Lactose Transport

scholarly journals Specificity Mutants of the Binding Protein of the Oligopeptide Transport System of Lactococcus lactis

2000 ◽  
Vol 182 (6) ◽  
pp. 1600-1608 ◽  
Author(s):  
Antonia Picon ◽  
Edmund R. S. Kunji ◽  
Frank C. Lanfermeijer ◽  
Wil N. Konings ◽  
Bert Poolman
Keyword(s):  
Lactococcus Lactis ◽  
Fold Increase ◽  
Binding Activity ◽  
Kinetic Properties ◽  
Wild Type ◽  
Mutant Proteins ◽  
Serovar Typhimurium ◽  
Peptide Binding Site ◽  
Oligopeptide Transport

ABSTRACT The kinetic properties of wild-type and mutant oligopeptide binding proteins of Lactococcus lactis were determined. To observe the properties of the mutant proteins in vivo, the oppAgene was deleted from the chromosome of L. lactis to produce a strain that was totally defective in oligopeptide transport. Amplified expression of the oppA gene resulted in an 8- to 12-fold increase in OppA protein relative to the wild-type level. The amplified expression was paralleled by increased bradykinin binding activity, but had relatively little effect on the overall transport of bradykinin via Opp. Several site-directed mutants were constructed on the basis of a comparison of the primary sequences of OppA fromSalmonella enterica serovar Typhimurium and L. lactis, taking into account the known structure of the serovar Typhimurium protein. Putative peptide binding-site residues were mutated. All the mutant OppA proteins exhibited a decreased binding affinity for the high-affinity peptide bradykinin. Except for OppA(D471R), the mutant OppA proteins displayed highly defective bradykinin uptake, whereas the transport of the low-affinity substrate KYGK was barely affected. Cells expressing OppA(D471R) had a similarKm for transport, whereas theV max was increased more than twofold as compared to the wild-type protein. The data are discussed in the light of a kinetic model and imply that the rate of transport is determined to a large extent by the donation of the peptide from the OppA protein to the translocator complex.

Evidence for two different broad-specificity oligopeptide transporters in intestinal cell line Caco-2 and colonic cell line CCD841

2011 ◽  
Vol 300 (6) ◽  
pp. C1260-C1269 ◽  
Author(s):  
Paresh Chothe ◽  
Nagendra Singh ◽  
Vadivel Ganapathy
Keyword(s):  
Amino Acids ◽  
Cell Line ◽  
Mammalian Cells ◽  
Peptide Transport ◽  
Transport Systems ◽  
Intestinal Cell ◽  
Small Peptides ◽  
Intestinal Cell Line ◽  
Deltorphin Ii ◽  
Oligopeptide Transport

Recently the existence of two different Na+-coupled oligopeptide transport systems has been described in mammalian cells. These transport systems are distinct from the previously known H+/peptide cotransporters PEPT1 and PEPT2, which transport only dipeptides and tripeptides. To date, the only peptide transport system known to exist in the intestine is PEPT1. Here we investigated the expression of the Na+-coupled oligopeptide transporters in intestinal cell lines, using the hydrolysis-resistant synthetic oligopeptides deltorphin II and [d-Ala2,d-Leu5]enkephalin (DADLE) as model substrates. Caco-2 cells and CCD841 cells, both representing epithelial cells from human intestinal tract, were able to take up these oligopeptides. Uptake of deltorphin II was mostly Na+ dependent, with more than 2 Na+ involved in the uptake process. In contrast, DADLE uptake was only partially Na+ dependent. The uptake of both peptides was also influenced by H+ and Cl−, although to a varying degree. The processes responsible for the uptake of deltorphin II and DADLE could be differentiated not only by their Na+ dependence but also by their modulation by small peptides. Several dipeptides and tripeptides stimulated deltorphin II uptake but inhibited DADLE uptake. These modulating small peptides were, however, not transportable substrates for the transport systems that mediate deltorphin II or DADLE uptake. These two oligopeptide transport systems were also able to take up several nonopioid oligopeptides, consisting of 9–17 amino acids. This represents the first report on the existence of transport systems in intestinal cells that are distinct from PEPT1 and capable of transporting oligopeptides consisting of five or more amino acids.

scholarly journals Specificity of peptide transport systems in Lactococcus lactis: evidence for a third system which transports hydrophobic di- and tripeptides.

1995 ◽  
Vol 177 (16) ◽  
pp. 4652-4657 ◽  
Author(s):  
C Foucaud ◽  
E R Kunji ◽  
A Hagting ◽  
J Richard ◽  
W N Konings ◽  
...  
Keyword(s):  
Lactococcus Lactis ◽  
Peptide Transport ◽  
Transport Systems

scholarly journals Transcriptional Pattern of Genes Coding for the Proteolytic System of Lactococcus lactis and Evidence for Coordinated Regulation of Key Enzymes by Peptide Supply

2001 ◽  
Vol 183 (12) ◽  
pp. 3614-3622 ◽  
Author(s):  
Eric Guédon ◽  
Pierre Renault ◽  
S. Dusko Ehrlich ◽  
Christine Delorme
Keyword(s):  
Lactococcus Lactis ◽  
Defined Medium ◽  
Transport Systems ◽  
Luciferase Reporter ◽  
Catabolic Repression ◽  
Proteolytic System ◽  
Transcriptional Pattern ◽  
Genes Encoding ◽  
Highly Expressed Genes ◽  
Responsive Element

ABSTRACT The transcription of 16 genes encoding 12 peptidases (pepC, pepN, pepX, pepP, pepA, pepF2, pepDA1, pepDA2, pepQ, pepT, pepM, and pepO1), PI and PIII proteinases (prtP1 and prtP3), and three transport systems (dtpT, dtpP, and opp-pepO1) ofLactococcus lactis MG1363 was analyzed in response to different environmental factors. Promoter fusions with luciferase reporter genes and/or mRNA analysis were used to study the effects of sugar sources, growth at 37°C, and peptide supply on the transcription of these genes. Only transcription of thepepP gene is modulated by the source of sugar. The presence of potential catabolite-responsive element (CRE) boxes in its promoter region suggests that expression of this gene is directly controlled by catabolic repression. Elevated temperature had no significant effect on the level of transcription of these genes. prtP1, prtP3, pepC, pepN, pepX, and the opp-pepO1 operon are the most highly expressed genes in chemically defined medium, and their expression is repressed 5- to 150-fold by addition of peptide sources such as Casitone in the medium. Moreover, the transcription ofprtP1, prtP3, pepC, pepN, and the opp-pepO1operon is repressed two- to eight-fold by the dipeptides leucylproline and prolylleucine. The transcription of pepDA2 might also be repressed by the peptide sources, but this effect is not observed on the regulation of dtpT, pepP, pepA, pepF2, pepDA1, pepQ, pepT, pepM, and the dtpP operon. The significance of these results with respect to the functions of different components of the proteolytic system in L. lactis are discussed.

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