{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T13:35:46Z","timestamp":1788960946175,"version":"build-2803163510"},"reference-count":52,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2020,5,10]],"date-time":"2020-05-10T00:00:00Z","timestamp":1589068800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100009872","name":"Regione Abruzzo","doi-asserted-by":"publisher","award":["MISURA 1.2.4."],"award-info":[{"award-number":["MISURA 1.2.4."]}],"id":[{"id":"10.13039\/501100009872","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>The aim of the study was to evaluate the proteolytic process in Caciocavallo cheese obtained from Friesian cows fed zinc, selenium, and iodine supplementation. Thirty-six Friesian cows, balanced for parity, milk production, and days in milk, were randomly assigned to four groups. The control group (CG) was fed with a conventional feeding strategy, while the three remaining groups received a diet enriched with three different trace elements, respectively zinc (ZG), selenium (SG), and iodine (IG). At the end of the experimental period, samples of milk were collected and used to produce Caciocavallo cheese from each experimental group. Cheese samples were then analyzed after 7 and 120 days from the cheese making in order to obtain information on chemical composition and extent of the proteolytic process, evaluated through the electrophoretic analysis of caseins and the determination of volatiles profile. Both milk and cheese samples were richer in the amount of the microelement respectively used for the integration of the cattle\u2019s diet. The zymographic approach was helpful in evaluating, in milk, the proteolytic function performed by endogenous metalloenzymes specifically able to degrade gelatin and casein; this evaluation did not highlight significant differences among the analyzed samples. In cheese, the electrophoretic analysis in reducing and denaturing condition showed the marked ability of \u03b2-casein to resist the proteolytic action during ripening, whereas the dietary selenium supplementation was shown to perform a protective action against the degradation of S1 and S2 isoforms of \u03b1-casein. The analysis of the volatile profile evidenced the presence of compounds associated with proteolysis of phenylalanine and leucine. This approach showed that selenium was able to negatively influence the biochemical processes that lead to the formation of 3-methyl butanol, although the identification of the specific mechanism needs further investigation.<\/jats:p>","DOI":"10.3390\/molecules25092249","type":"journal-article","created":{"date-parts":[[2020,5,12]],"date-time":"2020-05-12T02:41:20Z","timestamp":1589251280000},"page":"2249","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Proteolytic Volatile Profile and Electrophoretic Analysis of Casein Composition in Milk and Cheese Derived from Mironutrient-Fed Cows"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3102-6804","authenticated-orcid":false,"given":"Andrea","family":"Ianni","sequence":"first","affiliation":[{"name":"Faculty of BioScience and Technology for Food, Agriculture and Environment, University of Teramo, Via Renato Balzarini 1, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9030-4881","authenticated-orcid":false,"given":"Francesca","family":"Bennato","sequence":"additional","affiliation":[{"name":"Faculty of BioScience and Technology for Food, Agriculture and Environment, University of Teramo, Via Renato Balzarini 1, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Camillo","family":"Martino","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise \u201cG. Caporale\u201d, Via Campo Boario 37, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa","family":"Grotta","sequence":"additional","affiliation":[{"name":"Faculty of BioScience and Technology for Food, Agriculture and Environment, University of Teramo, Via Renato Balzarini 1, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicola","family":"Franceschini","sequence":"additional","affiliation":[{"name":"Department of Biotechnological and Applied Clinical Sciences, University of L\u2019Aquila, Via Vetoio 1, 67100 L\u2019Aquila, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7878-9318","authenticated-orcid":false,"given":"Giuseppe","family":"Martino","sequence":"additional","affiliation":[{"name":"Faculty of BioScience and Technology for Food, Agriculture and Environment, University of Teramo, Via Renato Balzarini 1, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,5,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1093\/jn\/118.2.229","article-title":"Effect of dietary selenium on the primary and secondary immune response in calves challenged with infectious bovine rhinotracheitis virus","volume":"118","author":"Reffett","year":"1988","journal-title":"J. Nutr."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1016\/S1367-5931(98)80064-1","article-title":"Zinc enzymes","volume":"2","author":"Coleman","year":"1998","journal-title":"Curr. Opin. Chem. Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.3168\/jds.S0022-0302(70)86355-X","article-title":"Zinc nutrition of cattle: A review","volume":"53","author":"Miller","year":"1970","journal-title":"J. Dairy Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/j.abb.2015.06.024","article-title":"Selenoproteins: Antioxidant selenoenzymes and beyond","volume":"595","author":"Steinbrenner","year":"2016","journal-title":"Arch Biochem. Biophys."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1016\/j.livsci.2009.07.005","article-title":"Effects of selenium yeast on rumen fermentation, lactation performance and feed digestibilities in lactating dairy cows","volume":"126","author":"Wang","year":"2009","journal-title":"Livestock Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1007\/s00394-013-0597-4","article-title":"Influencing factors on iodine content of cow milk","volume":"53","author":"Flachowsky","year":"2014","journal-title":"Eur. J. Nutr."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"745","DOI":"10.1017\/S1751731119002477","article-title":"Whole-transcriptome profiling of sheep fed with a high iodine-supplemented diet","volume":"14","author":"Iannaccone","year":"2019","journal-title":"Animal"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/j.anifeedsci.2017.10.007","article-title":"Transcriptome profiling and functional analysis of sheep fed with high zinc-supplemented diet: A nutrigenomic approach","volume":"234","author":"Elgendy","year":"2017","journal-title":"Anim. Feed Sci. Technol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Biscarini, F., Palazzo, F., Castellani, F., Masetti, G., Grotta, L., Cichelli, A., and Martino, G. (2018). Rumen microbiome in dairy calves fed copper and grape-pomace dietary supplementations: Composition and predicted functional profile. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0205670"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1016\/j.livsci.2009.12.005","article-title":"Effects of either sodium selenite or Se yeast (Sc CNCM I-3060) supplementation on selenium status and milk characteristics in dairy cows","volume":"128","author":"Calamari","year":"2010","journal-title":"Livestock Sci."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Iannaccone, M., Ianni, A., Elgendy, R., Martino, C., Giantin, M., Cerretani, L., Dacasto, M., and Martino, G. (2019). Iodine Supplemented Diet Positively Affect Immune Response and Dairy Product Quality in Fresian Cow. Animals, 9.","DOI":"10.3390\/ani9110866"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Schirone, M., Tofalo, R., Perpetuini, G., Manetta, A.C., Di Gianvito, P., Tittarelli, F., Battistelli, N., Corsetti, A., Suzzi, G., and Martino, G. (2018). Influence of iodine feeding on microbiological and physico-chemical characteristics and biogenic amines content in a raw ewes\u2019 milk cheese. Foods, 7.","DOI":"10.3390\/foods7070108"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/S0958-6946(01)00132-7","article-title":"Flavour formation from amino acids by lactic acid bacteria: Predictions from genome sequence analysis","volume":"12","author":"Kleerebezem","year":"2002","journal-title":"Int. Dairy J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1051\/lait:2000127","article-title":"Biochemical pathways for the production of flavour compounds in cheeses during ripening: A review","volume":"80","author":"McSweeney","year":"2000","journal-title":"Lait"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/S0958-6946(01)00049-8","article-title":"Cheese flavor formation by amino acid catabolism","volume":"11","author":"Yvon","year":"2001","journal-title":"Int. Dairy J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2918","DOI":"10.3168\/jds.2018-15868","article-title":"Zinc supplementation of Friesian cows: Effect on chemical-nutritional composition and aromatic profile of dairy products","volume":"102","author":"Ianni","year":"2019","journal-title":"J. Dairy Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/j.idairyj.2017.03.004","article-title":"Selenium supplementation of diets of dairy cows to produce Se-enriched cheese","volume":"71","author":"Ling","year":"2017","journal-title":"Int. Dairy. J."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"355","DOI":"10.2754\/avb200675030355","article-title":"Zinc supplementation and somatic cell count in milk of dairy cows","volume":"75","author":"Pavlata","year":"2006","journal-title":"Acta Vet. Brno"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1017\/S175173110999098X","article-title":"Iodine and selenium carry over in milk and cheese in dairy cows: Effect of diet supplementation and milk yield","volume":"4","author":"Moschini","year":"2010","journal-title":"Animal"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1007\/s11694-014-9203-6","article-title":"Heavy metals and trace elements levels in milk and milk products","volume":"8","author":"Meshref","year":"2014","journal-title":"J. Food Meas. Charact."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1016\/j.cardiores.2005.12.002","article-title":"Structure and function of matrix metalloproteinases and TIMPs","volume":"69","author":"Nagase","year":"2006","journal-title":"Cardiovasc. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1006\/abio.1993.1572","article-title":"An assay for matrix metalloproteinases and other proteases acting on proteoglycans, casein, or gelatin","volume":"215","author":"Manicourt","year":"1993","journal-title":"Anal. Biochem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.idairyj.2018.07.020","article-title":"Protease activity and protein profile in milk from healthy dairy cows and cows with different types of mastitis","volume":"89","author":"Caggiano","year":"2019","journal-title":"Int. Dairy J."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"660","DOI":"10.1111\/j.1365-2842.2004.01297.x","article-title":"Inhibition of human pulpal gelatinases (MMP-2 and MMP-9) by zinc oxide cements","volume":"31","author":"Santos","year":"2004","journal-title":"J. Oral Rehabil."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1666","DOI":"10.1039\/C3AY41706A","article-title":"Microfluidic chip electrophoresis investigation of major milk proteins: Study of buffer effects and quantitative approaching","volume":"6","author":"Costa","year":"2014","journal-title":"Anal. Met."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3066","DOI":"10.1021\/jf9802272","article-title":"Hydrolysis of Bovine and Caprine Caseins by Rennet and Plasmin in Model Systems","volume":"46","author":"Trujillo","year":"1998","journal-title":"J. Agric. Food Chem."},{"key":"ref_27","first-page":"E115","article-title":"Proteolytic and Rheological Properties of Aging Cheddar-Like Caprine Milk Cheeses Manufactured at Different Times during Lactation","volume":"72","author":"Tunick","year":"2007","journal-title":"J. Food Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jnutbio.2007.02.005","article-title":"Selenium as an anticancer nutrient: Roles in cell proliferation and tumor cell invasion","volume":"19","author":"Zeng","year":"2008","journal-title":"J. Nutr. Biochem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2166","DOI":"10.1016\/j.lwt.2011.05.023","article-title":"Beta casein-micelle as a nano vehicle for solubility enhancement of curcumin; food industry application","volume":"44","author":"Esmaili","year":"2011","journal-title":"LWT\u2014Food Sci. Technol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Ianni, A., Bennato, F., Martino, C., Grotta, L., and Martino, G. (2020). Volatile Flavor Compounds in Cheese as Affected by Ruminant Diet. Molecules, 25.","DOI":"10.3390\/molecules25030461"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1382","DOI":"10.1021\/jf001121l","article-title":"Characteristic aroma components of British farmhouse Cheddar cheese","volume":"49","author":"Suriyaphan","year":"2001","journal-title":"J. Agric. Food Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1007\/s00253-003-1447-8","article-title":"Diversity of L-leucine catabolism in various microorganisms involved in dairy fermentations, and identification of the rate-controlling step in the formation of the potent flavour component 3-methylbutanal","volume":"64","author":"Smit","year":"2004","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1051\/lait:2001002","article-title":"Production of cheese flavour compounds derived from amino acid catabolism by Propionibacterium freudenreichii","volume":"82","author":"Thierry","year":"2002","journal-title":"Lait"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2113","DOI":"10.1007\/s00217-019-03322-z","article-title":"Dietary selenium intake in lactating dairy cows modifies fatty acid composition and volatile profile of milk and 30-day-ripened caciotta cheese","volume":"245","author":"Ianni","year":"2019","journal-title":"Eur. Food Res. Technol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Martino, C., Ianni, A., Grotta, L., Pomilio, F., and Martino, G. (2019). Influence of Zinc Feeding on Nutritional Quality, Oxidative Stability and Volatile Profile of Fresh and Ripened Ewes\u2019 Milk Cheese. Foods, 8.","DOI":"10.3390\/foods8120656"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.idairyj.2019.02.014","article-title":"Zinc supplementation of dairy cows: Effects on chemical composition, nutritional quality and volatile profile of Giuncata cheese","volume":"94","author":"Ianni","year":"2019","journal-title":"Int. Dairy J."},{"key":"ref_37","unstructured":"(2020, March 25). European Union (2010) Directive 2010\/63\/EU of the European parliament and of the council of 22 September 2010 on the protection of animals used for scientific purposes. Available online: https:\/\/eur-lex.europa.eu\/legal-content\/EN\/TXT\/PDF\/?uri=CELEX:32010L0063&from=EN."},{"key":"ref_38","unstructured":"(2020, March 25). European Economic Community (1986) Council Directive 86\/609\/EEC of 24 November 1986 on the approximation of laws, regulations and administrative provisions of the Member States regarding the protection of animals used for experimental and other scientific purposes. Available online: https:\/\/eur-lex.europa.eu\/legal-content\/EN\/TXT\/PDF\/?uri=CELEX:31986L0609&from=EN."},{"key":"ref_39","unstructured":"(2020, March 25). European Commission (2003) Regulation (EC) No. 1831\/2003 of the European Parliament and of the Council of 22 September 2003 on Additives for Use in Animal Nutrition. Available online: https:\/\/eur-lex.europa.eu\/legal-content\/EN\/TXT\/PDF\/?uri=CELEX:32003R1831&from=EN."},{"key":"ref_40","unstructured":"(2020, March 25). European Commission (2014) Regulation No. 121\/2014 of the concerning the authorization of L-selenomethionine as a feed additive for all animal species. Available online: https:\/\/eur-lex.europa.eu\/legal-content\/EN\/TXT\/PDF\/?uri=CELEX:32014R0121&from=EN."},{"key":"ref_41","unstructured":"(2020, March 25). European Commission (2005) Regulation No. 1459\/2005 amending the conditions for authorisation of a number of feed additives belonging to the group of trace elements. Available online: https:\/\/eur-lex.europa.eu\/LexUriServ\/LexUriServ.do?uri=OJ:L:2005:233:0008:0010:EN:PDF."},{"key":"ref_42","unstructured":"NRC (2001). Nutrient Requirements of Dairy Cattle, Natl Acad Press."},{"key":"ref_43","unstructured":"AOAC (1990). Official Methods of Analysis, Association of Official Analytical Chemists. [15th ed.]."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3583","DOI":"10.3168\/jds.S0022-0302(91)78551-2","article-title":"Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition","volume":"74","author":"Robertson","year":"1991","journal-title":"J. Dairy Sci."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"6853","DOI":"10.3168\/jds.2019-16382","article-title":"Effects of selenium supplementation on chemical composition and aromatic profiles of cow milk and its derived cheese","volume":"102","author":"Ianni","year":"2019","journal-title":"J. Dairy Sci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"912","DOI":"10.1016\/j.talanta.2004.04.004","article-title":"Direct determination of Cu and Zn in fruit juices and bovine milk by thermospray flame furnace atomic absorption spectrometry","volume":"64","author":"Nascentes","year":"2004","journal-title":"Talanta"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1017\/S1751731108003212","article-title":"Variability of selected trace elements of different meat cuts determined by ICP-MS and DRC-ICPMS","volume":"3","author":"Gerber","year":"2009","journal-title":"Animal"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"977","DOI":"10.1039\/a801671b","article-title":"Determination of iodine in food samples by inductively coupled plasma mass spectrometry after alkaline extraction","volume":"13","author":"Fecher","year":"1998","journal-title":"J. Anal. At. Spectrom."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Kruger, N.J. (2009). The Bradford method for protein quantitation. The Protein Protocols Handbook, Humana Press.","DOI":"10.1007\/978-1-59745-198-7_4"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1111\/cbdd.13468","article-title":"Oxaprozin: A new hope in the modulation of matrix metalloproteinase 9 activity","volume":"93","author":"Ianni","year":"2019","journal-title":"Chem. Biol. Drug Des."},{"key":"ref_51","unstructured":"Rasband, W.S. (1997). ImageJ Software, National Institutes of Health."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"148","DOI":"10.5713\/ajas.18.0868","article-title":"Chemical-nutritional characteristics and aromatic profile of milk and related dairy products obtained from goats fed with extruded linseed","volume":"33","author":"Bennato","year":"2020","journal-title":"Asian-Australas. J. Anim. Sci."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/25\/9\/2249\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:27:26Z","timestamp":1760174846000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/25\/9\/2249"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,5,10]]},"references-count":52,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2020,5]]}},"alternative-id":["molecules25092249"],"URL":"https:\/\/doi.org\/10.3390\/molecules25092249","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,5,10]]}}}