Proline-specific activities of<i>Propionibacterium freudenreichii</i>subsp.<i>shermanii</i>

Quelen, Laurent; Dupuis, C.; Boyaval, Patrick

doi:10.1017/s0022029900031411

Cited by 9 publications

(9 citation statements)

References 14 publications

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“…shermanii CIP103027 strain (approximately 3 and 2.2 amino acid residues, respectively), and consequently their release is low. With regard to proline, such a low re1ease from the tryptic/chymotryptic~-casein peptides can be attributed to its central or ante-penultimate position in the peptide sequences [26], which perhaps should require previous proteolysis by other aminopeptidases for being released by the very specifie enzyme activities already shown in propionibacteria (proline iminopeptidase [25,27], prolinase and prolidase [29]), and X-prolyl dipeptidyl aminopeptidase [13]). …”

Section: Discussionmentioning

confidence: 99%

“…They are mainly intracellularly located [12,23,27,31]. Four of them identified in P. freudenreichii are supposed to play a major role in Swiss cheese flavor development: a proline iminopeptidase [25,27], prolinase and prolidase [29] and two phenylalanine aminopeptidases [10]. The amino acids, Pro and Phe, are released at a high level during ripening and shown respectively as component participating to the sweetness and as a precursor of floral flavor of the cheese [3,6,17,18].…”

Section: Dairy Propionibacteriamentioning

confidence: 99%

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Strain variability of the cell-free proteolytic activity of dairy propionibacteria towards β-casein peptides

Lemée

Gagnaire

Maubois

1998

Lait

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-Strain selection of dairy propionibacteria is based on COz' acetate and propionate production. However, another criterion such as proteolytic activity could be of industrial interest for selection of flavor ripening agents. Given that the propionibacteria are variously able to autolyse and that proteolytic activity is mainly intracellularly located, the proteolytic potential was evaluated on the cell-free extracts of 37 dairy propionibacteria strains. The substrate used to detect proteolytic activity was a tryptic/chymotryptic hydrolysate of~-casein. The evaluation was performed at a temperature of 30 "C and pH of 5.7. The increase in the free NH z groups over a 24 h period was monitored as weil as the evolution of the reversed-phase chromatographie profiles. In addition, for 14 strains, the free amino acids released from the~-casein peptides were determined. Ali the strains were able to hydrolyze~-casein peptides. Difference of proteolytic activity was observed among and within the four dairy propionibacteria species. This variability was expressed more in terms of nature of the products released (peptides and free amino acids) than in terms of degree of proteolytic activity. From the results, a strain selection system may be developed using as substrate the tryptic/chymotryptic~-casein hydrolysate and as criteria the degree of hydrolysis at 24 h and the amount and the nature of free ami no acids released by cellfree ex tracts. Such a selection will give useful data for cheesemakers, especially if characterization of autolytic ability in the cheese environment is also determined. © InralElsevier, Paris.propionibacteria / proteolysis / peptide / Cree amino acid / selection Résumé -Souche dépendance de l'activité protéolytique intracellulaire de bactéries propioniques laitières sur un substrat laitier (hydrolysat de caséine~). La sélection des souches de bactéries propioniques est fondée sur la production de COz' d'acétate et de propionate. Cependant, un autre critère tel que l'activité protéolytique pourrait présenter un intérêt industriel afin de sélectionner des souches susceptibles de produire des précurseurs de flaveurs. En tenant compte du fait que les bactéries propioniques possèdent une capacité variable à s'autolyser et que * Correspondence and reprints. E-mail: gagnaire@labtechno.roazhon.inra.fr of intracellular peptidase activities in cheese curd during ripening requires evidently an autolytic mechanism induced in propionibacteria cells, either physiologically or by environmental conditions. Such an autolysis was recently observed under in vitro conditions [20, 21, 23, 24] and in Emmental (Valence F., personal communication). l'activité protéolytique est localisée principalement dans la fraction cytoplasmique de la bacté-rie, nous avons évalué le potentielprotéolytiqued'extrait intracellulairede 37 souchesde bactéries propioniques. Le substrat utilisé pour tester l'activité de manière globale était un hydrolysat trypsique/chymotrypsique de caséine~. L'hydrolyse était réalisée à 30 "C et à pH...

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Section: Discussionmentioning

confidence: 99%

Section: Dairy Propionibacteriamentioning

confidence: 99%

Strain variability of the cell-free proteolytic activity of dairy propionibacteria towards β-casein peptides

Lemée

Gagnaire

Maubois

1998

Lait

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“…Sebastiani and Tschager [82] also observed the role of aspartate metabolism in succinate production in Swiss-type cheese. PAB are equipped with a complex intracellular peptidase system [43,68] and there is strong evidence that PAB can utilise short chain peptides [4,64,65]. It is likely that amino acids and peptides, produced by proteolysis during ripening form a pool sorne of which are subsequently used by PAB; the extent of this utilisation may explain at least sorne of the variations in the relative concentrations of propionate, acetate, CO 2 and succinate produced.…”

Section: Fermentation Of Substra Te Mixturesmentioning

confidence: 99%

Metabolism of lactate and sugars by dairy propionibacteria: A review

Piveteau¹

1999

Lait

106

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-Dairy propionibacteria are important organisms for the manufacture of Swiss-type cheese, for the biological production of propionate and vitamin 8 12 and have probiotic properties. In ail the se applications, their metabolic activities play a critical role. A complete understanding of propionate fermentation and of the metabolic routes used is therefore necessary. Dairy propionibacteria have a complex metabolism and involves several cycles. Lactate or sugars utilisation yields pyruvate which can be reduced to produce propionate via the transcarboxylase cycles, or oxidised to yield acetate and CO 2 • During the coupled oxidation-reduction, ATP is produced by an electron transport system and fumarate acts as the final acceptor. Although propionibacteria are mainly anaerobes, the electron transport system can be used in the presence of oxygen and they possess the citrate cycle, but can not grow under normal atmospheric oxygen pressure. The proportions of propionate acetate and CO 2 produced vary depending on the strain used and this can be explained, to sorne extent, by their relative ability to utilise pyruvate via reactions of the citrate cycle. The physicochemical environ ment during growth affects propionic acid fermentation; it is impaired by the presence of oxygen and nitrate, but fermentation at acidic pHs enhances propionate production. Fermentation in presence of more than one substrate is complex and still poorly understood. When both L-and D-lactate isomers are available, L-lactate is used preferentially. Although sugar fermentation is more efficient, in the presence of lactate and sugars, there is evidence that lactate is used preferentially. Propionic acid fermentation is affected by the utilisation of amino acids, especially aspartate; co-metabolism of lactate and aspartate results in a lower propionate production and a decrease of the ratio propionate:acetate. There is evidence that the utilisation of the products of proteolysis is an important event in the ripening of Swiss-type chee se and could account for the low propionate:acetate ratios observed in Swiss-type cheese. © Inra/Elsevier, Paris.propionic acid bacteria {lactate { sugar { metabolism Résumé -Métabolisme du lactate et des sucres par les bactéries propioniques laitières: une revue. Les bactéries propioniques laitières sont importantes pour la fabrication de fromages de type emmental, pour la production biologique de propionate et de vitamine 8 12 ainsi que pour leurs propriétés probiotiques. Dans ces domaines, leurs activités métaboliques ont un rôle crucial. Il est donc Oral communication at the 2nd Symposium on Propionibacteria, Cork, Ireland, June 25-27, 1998. * Correspondence and reprints. P.Piveteau@hotmail.com 24 P. Piveteau nécessaire de comprendre de façon approfondie la fermentation propionique et les voies métabo-liques empruntées. Les bactéries propioniques possèdent un métabolisme complexe impliquant plusieurs cycles. Le pyruvate issu de l'utilisation du lactate et des sucres peut être réduit, ce qui conduit à la pr...

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“…These observations are in contrast to recent work by Valence et al (46), in which autolysis of P. freudenreichii was reported to occur to a limited extent and very late during cold storage of Swiss cheese. Research on proteolysis by propionibacteria has mainly focused on free proline and intracellular proline-specific peptidase activity, and release of peptidases into cheese through autolysis has been assumed (11,23,27,33,43). Langsrud et al (21,22) observed a correlation between autolysis of propionibacteria in milk and production of free proline, and Østlie et al (31) confirmed this correlation for broth media.…”

Section: Resultsmentioning

confidence: 99%

Use of DNA Quantification To Measure Growth and Autolysis of Lactococcus and Propionibacterium spp. in Mixed Populations

Treimo

Vegarud

Langsrud

et al. 2006

Appl Environ Microbiol

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Autolysis is self-degradation of the bacterial cell wall that results in the release of enzymes and DNA. Autolysis of starter bacteria, such as lactococci and propionibacteria, is essential for cheese ripening, but our understanding of this important process is limited. This is mainly because the current tools for measuring autolysis cannot readily be used for analysis of bacteria in mixed populations. We have now addressed this problem by species-specific detection and quantification of free DNA released during autolysis. This was done by use of 16S rRNA gene single-nucleotide extension probes in combination with competitive PCR. We analyzed pure and mixed populations of Lactococcus lactis subsp. lactis and three different species of Propionibacterium. Results showed that L. lactis subsp. lactis INF L2 autolyzed first, followed by Propionibacterium acidipropionici ATCC 4965, Propionibacterium freudenreichii ISU P59, and then Propionibacterium jensenii INF P303. We also investigated the autolytic effect of rennet (commonly used in cheese production). We found that the effect was highly strain specific, with all the strains responding differently. Finally, autolysis of L. lactis subsp. lactis INF L2 and P. freudenreichii ISU P59 was analyzed in a liquid cheese model. Autolysis was detected later in this cheese model system than in broth media. A challenge with DNA, however, is DNA degradation. We addressed this challenge by using a DNA degradation marker. We obtained a good correlation between the degradation of the marker and the target in a model experiment. We conclude that our DNA approach will be a valuable tool for use in future analyses and for understanding autolysis in mixed bacterial populations.The present tools for measuring autolysis are not easily adaptable to analysis of mixed populations. This severely limits our understanding of autolysis in microbial populations, such as in cheese. Autolysis is self-degradation of the bacterial cell wall due to hydrolysis of the peptidoglycan network by autolysins (32,34,36,37), and the result is leakage of intracellular components (9, 27). Lactococcus lactis subsp. lactis and Propionibacterium freudenreichii are bacteria essential to the ripening of Norwegian Jarlsberg cheese (17). The propionibacteria produce propionic acid, acetic acid, and CO 2 , with the latter responsible for the typical eye formation. In addition, this secondary starter contributes to the characteristic sweet, nutty flavor, provided mainly by free proline (20)(21)(22). Autolysis of lactococci is important for cheese ripening, in which the release of intracellular enzymes plays an important role in cheese flavor development (9, 40, 48). The role and contribution of intracellular enzymes from propionibacteria in the ripening process are poorly understood (11,12,23,27,40). Proteolytic enzymes, such as chymotrypsin and pepsin, are reported to modify autolytic activity (28,29,38,39). Rennet, consisting of chymosin and bovine pepsin, hydrolyzes the -casein in milk and thus is a vital factor in ...

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Proline-specific activities ofPropionibacterium freudenreichiisubsp.shermanii

Cited by 9 publications

References 14 publications

Strain variability of the cell-free proteolytic activity of dairy propionibacteria towards β-casein peptides

Strain variability of the cell-free proteolytic activity of dairy propionibacteria towards β-casein peptides

Metabolism of lactate and sugars by dairy propionibacteria: A review

Use of DNA Quantification To Measure Growth and Autolysis of Lactococcus and Propionibacterium spp. in Mixed Populations

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