A Model to Assess Lactic Acid Bacteria Aminopeptidase Activities in Parmigiano Reggiano Cheese During Ripening
Open accessAbstract
The aim of this work was to investigate in which phases of ripening of Parmigiano Reggiano cheese lactic acid bacteria aminopeptidases present in cheese extract could be involved in release of free amino acids and to better understand the behavior of these enzymes in physical-chemical conditions that are far from their optimum. In particular, we evaluated 6 different substrates to reproduce broad-specificity aminopeptidase N, broad-specificity aminopeptidase C, glutamyl aminopeptidase A, peptidase with high specificity for leucine and alanine, proline iminopeptidase, and X-prolyl dipeptidyl aminopeptidase activities releasing different N-terminal amino acids. The effects of pH, NaCl concentration, and temperature on the enzyme activities of amino acid beta-naphthylamide (betaNA)-substrates were determined by modulating the variables in 19 different runs of an experimental design, which allowed the building of mathematical models able to assess the effect on aminopeptidases activities over a range of values, obtained with bibliographic data, covering different environmental conditions in different zones of the cheese wheel at different aging times. The aminopeptidases tested in this work were present in cell-free Parmigiano Reggiano cheese extract after a 17-mo ripening and were active when tested in model system. The modeling approach shows that to highlight the individual and interactive effects of chemical-physical variables on enzyme activities, it is helpful to determine the true potential of an amino-peptidase in cheese. Our results evidenced that the 6 different lactic acid bacteria peptidases participate in cheese proteolysis and are induced or inhibited by the cheese production parameters that, in turn, depend on the cheese dimension. Generally, temperature and pH exerted the more relevant effects on the enzymatic activities, and in many cases, a relevant interactive effect of these variables was observed. Increasing salt concentration slowed down broad-specificity amino-peptidase C, glutamyl aminopeptidase A, proline iminopeptidase, and peptidase with high specificity for leucine and alanine. Interestingly, this variable did not affect broad-specificity aminopeptidase N and positively affected X-prolyl dipeptidyl aminopeptidase. The models elaborated varying pH, temperatures, and salt concentration and were a useful, low cost, and fast tool to understand the role of the main peptidases in the different phases of cheese ripening in relation to the major environmental factors influencing enzyme activity.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
This large population of bacteria represents a significant reservoir of enzymatic activity because their breakdown and the release of intracellular enzymes are crucial factors in cheese ripening.
“This large population of bacteria represents a significant reservoir of enzymatic activity because their breakdown and the release of intracellular enzymes are crucial factors in cheese ripening [18,69,73,88,90,91,92,93,94,95,96,97,98].”
From Microbiota of Cheese Ecosystems: A Perspective on Cheesemaking (2025) · cites this paper as [92]
Cites
Addelman, S., Box, G. E. P., Hunter, W. G., & Hunter, J. S. (1979) · Technometrics · doi:10.2307/1267766 · Cited by 6,753 · see citing works
Flavour formation by lactic acid bacteria and biochemical flavour profiling of cheese products
Smit, G., Smit, B. A., & Engels, W. J. M. (2005) · FEMS Microbiology Reviews · doi:10.1016/j.fmrre.2005.04.002 · Cited by 949 · see citing works
Proteolytic systems of lactic acid bacteria.
Savijoki, K., Ingmer, H., & Varmanen, P. (2006) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-006-0427-1 · Cited by 717 · see citing works
The proteotytic systems of lactic acid bacteria
Kunji, E. R. S., Mierau, I., Hagting, A., Poolman, B., & Konings, W. N. (1996) · Antonie van Leeuwenhoek · doi:10.1007/bf00395933 · Cited by 715 · see citing works
Peptidases and amino acid catabolism in lactic acid bacteria
Christensen, J. E., Dudley, E. G., Pederson, J. A., & Steele, J. L. (1999) · Antonie van Leeuwenhoek · doi:10.1023/a:1002001919720 · Cited by 630 · see citing works
Amino Acid Catabolic Pathways of Lactic Acid Bacteria
Fernández, M., & Zúñiga, M. (2006) · Critical Reviews in Microbiology · doi:10.1080/10408410600880643 · Cited by 416 · see citing works
Autolysis and proteolysis in different strains of starter bacteria during Cheddar cheese ripening
Wilkinson, M. G., Guinee, T. P., O'Callaghan, D. M., & Fox, P. F. (1994) · Journal of Dairy Research · doi:10.1017/s0022029900028260 · Cited by 208 · see citing works
The proteolytic system of lactic acid bacteria
Kok, J., & De Vos, W. M. (1994) · Genetics and Biotechnology of Lactic Acid Bacteria · doi:10.1007/978-94-011-1340-3_4 · Cited by 205 · see citing works
Autolysis of two strains of Lactococcus lactis during cheese ripening
Chapot, M.-P., Deniel, C., Rousseau, M., Vassal, L., & Gripon, J. C. (1994) · International Dairy Journal · doi:10.1016/0958-6946(94)90016-7 · Cited by 143 · see citing works
Addeo, F., Chianese, L., Salzano, A., Sacchi, R., Cappuccio, U., Ferranti, P., & Malorni, A. (1992) · Journal of Dairy Research · doi:10.1017/s0022029900030673 · Cited by 143 · see citing works
Plasmin System and Microbial Proteases in Milk: Characteristics, Roles, and Relationship
Nielsen, S. S. (2002) · Journal of Agricultural and Food Chemistry · doi:10.1021/jf0201881 · Cited by 111 · see citing works
Crow, V. L., Martley, F. G., Coolbear, T., & Roundhill, S. J. (1995) · International Dairy Journal · doi:10.1016/0958-6946(95)00022-u · Cited by 79 · see citing works
Chemical parameters of the non-volatile fraction of ripened Parmigiano-Reggiano cheese
Careri, M., Spagnoli, S., Panari, G., Zannoni, M., & Barbieri, G. (1996) · International Dairy Journal · doi:10.1016/0958-6946(94)00056-5 · Cited by 53 · see citing works
PepS from Streptococcus thermophilus
Fernández-Esplá, M. D., & Rul, F. (1999) · European Journal of Biochemistry · doi:10.1046/j.1432-1327.1999.00528.x · Cited by 51 · see citing works
Proteolysis of β-Casein as a Marker of Grana Padano Cheese Ripening
Gaiaschi, A., Beretta, B., Poiesi, C., Conti, A. M. F., Giuffrida, M. G., Galli, C. L., & Restani, P. (2001) · Journal of Dairy Science · doi:10.3168/jds.s0022-0302(01)74452-9 · Cited by 51 · see citing works
Ferranti, P., Itolli, E., Barone, F., Malorni, A., Garro, G., Laezza, P., Chianese, L., Migliaccio, F., Stingo, V., & Addeo, F. (1997) · Le Lait · doi:10.1051/lait:1997649 · Cited by 50 · see citing works
Effects of Heat Load Gradient Occurring in Moulding on Characterization and Ripening of Grana Padano
Pellegrino, L., Battelli, G., Resmini, P., Ferranti, P., Barone, F., & Addeo, F. (1997) · Le Lait · doi:10.1051/lait:1997215 · Cited by 47 · see citing works
Mucchetti, G., Locci, F., Gatti, M., Neviani, E., Addeo, F., Dossena, A., & Marchelli, R. (2000) · Journal of Dairy Science · doi:10.3168/jds.s0022-0302(00)74926-5 · Cited by 45 · see citing works
Self-citationGiraffa, G., Rossetti, L., Mucchetti, G., Addeo, F., & Neviani, E. (1998) · Journal of Dairy Science · doi:10.3168/jds.s0022-0302(98)75547-x · Cited by 45 · see citing works
Self-citationProteolytic specificity of cathepsin G on bovine α s1 - and β-caseins
Considine, T., Geary, S. J., Kelly, A. L., & McSweeney, P. L. H. (2002) · Food Chemistry · doi:10.1016/s0308-8146(01)00250-3 · Cited by 43 · see citing works
Varmanen, P., Vesanto, E., Steele, J. L., & Palva, A. (1994) · FEMS Microbiology Letters · doi:10.1111/j.1574-6968.1994.tb07302.x · Cited by 41 · see citing works
Klein, J. R., Dick, A., Schick, J., Matern, H. T., Henrich, B., & Plapp, R. (1995) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1995.tb20296.x · Cited by 34 · see citing works
Nucleotide sequence and distribution of the pepPN gene from Lactobacillus helveticus CNRZ32
Dudley, E. G., & Steele, J. L. (1994) · FEMS Microbiology Letters · doi:10.1111/j.1574-6968.1994.tb06864.x · Cited by 33 · see citing works
Free active peptidases are detected in Emmental juice extracted before ripening in the warm room
Gagnaire, V., Lortal, S., & Léonil, J. (1998) · Journal of Dairy Research · doi:10.1017/s0022029997002525 · Cited by 32 · see citing works
Proteolysis of αs-Casein as a Marker of Grana Padano Cheese Ripening
Gaiaschi, A., Beretta, B., Poiesi, C., Conti, A. M. F., Giuffrida, M. G., Galli, C. L., & Restani, P. (2000) · Journal of Dairy Science · doi:10.3168/jds.s0022-0302(00)75167-8 · Cited by 31 · see citing works
Boquien, C., Corrieu, G., & Desmazeaud, M. (1989) · Applied Microbiology and Biotechnology · doi:10.1007/bf00296631 · Cited by 25 · see citing works
Sforza, S., Galaverna, G., Neviani, E., Pinelli, C., Dossena, A., & Marchelli, R. (2004) · European Journal of Mass Spectrometry · doi:10.1255/ejms.603 · Cited by 24 · see citing works
Self-citationPresence of Peptidase Activities in Different Varieties of Cheese
Gatti, M., Fornasari, M. E., Mucchetti, G., Addeo, F., & Neviani, E. (1999) · Letters in Applied Microbiology · doi:10.1046/j.1365-2672.1999.00541.x · Cited by 23 · see citing works
Self-citationPeptidase activity in various species of dairy thermophilic lactobacilli
Gatti, M., Fornasari, M. E., Lazzi, C., Mucchetti, G., & Neviani, E. (2004) · Journal of Applied Microbiology · doi:10.1046/j.1365-2672.2003.02142.x · Cited by 21 · see citing works
Self-citationSt-PepA, a Streptococcus thermophilus aminopeptidase with high specificity for acidic residues
Rul, F., Gripon, J. C., & Monnet, V. (1995) · Microbiology · doi:10.1099/13500872-141-9-2281 · Cited by 20 · see citing works
Resmini, P., Pellegrino, L. M., Pazzaglia, C., & Hogenboom, J. A. (1985) · Cited by 13 · see citing works
Synthetic peptides as substrate for assaying the proteolytic activity of Lactobacillus helveticus
Caira, S., Ferranti, P., Gatti, M., Fornasari, M. E., Barone, F., Lilla, S., Mucchetti, G., Picariello, G., Chianese, L., Neviani, E., et al. (2003) · Journal of Dairy Research · doi:10.1017/s0022029903006368 · Cited by 11 · see citing works
Self-citationMichele, D. S. (2000) · CINECA IRIS Institutial research information system (University of Pisa) · Cited by 11 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Gatti, M., Bottari, B., Lazzi, C., Neviani, E., & Mucchetti, G. (2014) · Journal of Dairy Science · doi:10.3168/jds.2013-7187 · Cited by 216 · see citing works
Self-citationOlivares, M., Schüppel, V., Hassan, A. M., Beaumont, M., Neyrinck, A. M., Bindels, L. B., Benítez-Páez, A., Sanz, Y., Haller, D., Holzer, P., et al. (2018) · Frontiers in Microbiology · doi:10.3389/fmicb.2018.01900 · Cited by 72 · see citing works
Neviani, E., Bottari, B., Lazzi, C., & Gatti, M. (2013) · Frontiers in Microbiology · doi:10.3389/fmicb.2013.00036 · Cited by 66 · see citing works
Self-citationLazzi, C., Povolo, M., Locci, F., Bernini, V., Neviani, E., & Gatti, M. (2016) · International Journal of Food Microbiology · doi:10.1016/j.ijfoodmicro.2016.06.009 · Cited by 60 · see citing works
Self-citationAntioxidant capacity of water soluble extracts from Parmigiano-Reggiano cheese
Bottesini, C., Paolella, S., Lambertini, F., Galaverna, G., Tedeschi, T., Dossena, A., Marchelli, R., & Sforza, S. (2013) · International Journal of Food Sciences and Nutrition · doi:10.3109/09637486.2013.821696 · Cited by 44 · see citing works
Microbiota of Cheese Ecosystems: A Perspective on Cheesemaking
Neviani, E., Gatti, M., Gardini, F., & Levante, A. (2025) · Foods · doi:10.3390/foods14050830 · Cited by 39 · see citing works
Self-citationZago, M., Massimiliano, L., Bonvini, B., Penna, G., Giraffa, G., & Rescigno, M. (2021) · PLoS ONE · doi:10.1371/journal.pone.0245903 · Cited by 33 · see citing works
Comprehensive analysis of proteolysis during 8 months of ripening of high-cooked Old Saare cheese
Taivosalo, A., Kriščiunaite, T., Seiman, A., Part, N., Stulova, I., & Vilu, R. (2017) · Journal of Dairy Science · doi:10.3168/jds.2017-12944 · Cited by 30 · see citing works
Moatsou, G. A., Moschopoulou, E., Beka, A., Tsermoula, P., & Pratsis, D. (2015) · International Dairy Journal · doi:10.1016/j.idairyj.2015.05.010 · Cited by 26 · see citing works
Li, N., Wu, J., Zhang, L.-F., Zhang, Y., & Feng, H. (2010) · Biochimie · doi:10.1016/j.biochi.2010.02.022 · Cited by 26 · see citing works
Merchán, A. V., Ruiz‐Moyano, S., Vázquez-Hernández, M., Benito, M. J., Aranda, E., Rodríguez, A., & Martı́n, A. (2022) · Journal of Dairy Science · doi:10.3168/jds.2021-21368 · Cited by 20 · see citing works
Søndergaard, L., Ryssel, M., Svendsen, C., Høier, E., Andersen, U., Hammershøj, M., Møller, J. R., Arneborg, N., & Jespersen, L. (2015) · International Journal of Food Microbiology · doi:10.1016/j.ijfoodmicro.2015.06.031 · Cited by 20 · see citing works
Neviani, E., Levante, A., & Gatti, M. (2024) · Fermentation · doi:10.3390/fermentation10040186 · Cited by 17 · see citing works
Self-citationMilesi, M. M., Bergamini, C. V., & Hynes, E. R. (2011) · Food Research International · doi:10.1016/j.foodres.2011.01.007 · Cited by 13 · see citing works
Bettera, L., Alinovi, M., Mondinelli, R., & Mucchetti, G. (2020) · Foods · doi:10.3390/foods9081101 · Cited by 12 · see citing works
Self-citationCelano, G., Costantino, G., Calasso, M. P., Randazzo, C. L., & Minervini, F. (2022) · Foods · doi:10.3390/foods11030425 · Cited by 10 · see citing works
The Most Traditional and Popular Italian Cheeses
Gobbetti, M., Neviani, E., & Fox, P. F. (2018) · Springer eBooks · doi:10.1007/978-3-319-89854-4_6 · Cited by 8 · see citing works
Self-citationValentino, V., Magliulo, R., Balivo, A., Krysmann, A., Calvanese, C. M., Esposito, A., Sequino, G., Genovese, A., Porcellato, D., Ercolini, D., et al. (2025) · npj Biofilms and Microbiomes · doi:10.1038/s41522-025-00815-6 · Cited by 8 · see citing works
Technological and functional potentials of indigenous yeasts from traditional Tulum cheese
Aydın, F. (2025) · World Journal of Microbiology and Biotechnology · doi:10.1007/s11274-025-04668-1 · Cited by 6 · see citing works
Short communication: A new minicurd model system for hard cooked cheeses
Vélez, M. A., Perotti, M. C., Rebechi, S. R., & Hynes, E. R. (2015) · Journal of Dairy Science · doi:10.3168/jds.2014-8906 · Cited by 6 · see citing works
Neviani, E. (2024) · Microorganisms · doi:10.3390/microorganisms12122443 · Cited by 5 · see citing works
Self-citationExploring Extremozymes: Adaptations, Bioprospecting, Enzyme Engineering and Applications
Alabhai, J. M., Manjula, B., Mishra, H., & Sangeetha, J. (2025) · Current Enzyme Inhibition · doi:10.2174/0115734080360789250414062505 · Cited by 2 · see citing works
Asim, S. M., Warakaulle, S., Ayyash, M. M., & Kamal‐Eldin, A. (2026) · Journal of Dairy Science · doi:10.3168/jds.2026-28766 · Cited by 2 · see citing works
Development of green brined cheese enriched with Chlorella vulgaris biomass
Hernández, H., Nunes, M. C., Prista, C., Moschopoulou, E., Sakkas, L., & Raymundo, A. (2026) · Algal Research · doi:10.1016/j.algal.2026.104977
- Authors
- Gatti, M., De Dea Lindner, J., Gardini, F., Mucchetti, G., Bevacqua, D., Fornasari, M. E., & Neviani, E.
- Affiliations
- University of Parma (IT)
- University of Bologna (IT)
- Foraggere e Lattiero Casearie (IT)
- Corresponding author
- Gatti, M.
- Venue
- Journal of Dairy Science
- Publisher
- American Dairy Science Association
- Published
- 2008-11
- Volume
- 91
- Issue
- 11
- Pages
- 4129-4137
- ISSN
- 0022-0302 (ISSN-L)
- OpenAlex
- W2043957138
- Cited by
- 24
- References
- 33
- Citations by year
- 2026: 2 · 2025: 4 · 2024: 2 · 2022: 2 · 2021: 1 · 2020: 1 · 2018: 2 · 2017: 1 · 2016: 1 · 2015: 3 · 2013: 3
- FWCI
- 1.07
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Journal of Dairy Science (publishedVersion)
- PubMed (publishedVersion)
- HAL (submittedVersion)
- Archivio istituzionale della ricerca (Alma Mater Studiorum Università di Bologna) (submittedVersion)
- INFM-OAR (INFN Catania) (submittedVersion, other-oa)
- Funders
- “SMEPR Studio e modellazione degli aspetti enzimatici legati alla stagionatura del formaggio Parmigiano-Reggiano”
- Emilia Romagna Region (LR28/98)
- Topics
- Life Sciences › Agricultural and Biological Sciences › Food Science › Probiotics and Fermented Foods
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Hydrolysis and Bioactive Peptides
- Keywords
- Parmigiano-Reggiano cheese, lactic acid bacteria, aminopeptidase, cheese ripening, cheese proteolysis, free amino acids, proline iminopeptidase, X-prolyl dipeptidyl aminopeptidase, pH, NaCl concentration, temperature
- MeSH terms
- Aminopeptidases, Cheese, Food Handling, Hydrogen-Ion Concentration, Lactobacillus, Models, Theoretical, Sodium Chloride, Temperature, Lactic Acid
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
