{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,20]],"date-time":"2026-06-20T05:25:24Z","timestamp":1781933124435,"version":"3.54.5"},"reference-count":58,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2024,3,12]],"date-time":"2024-03-12T00:00:00Z","timestamp":1710201600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Science and Technology Innovation Key Project of China Light Industry Group (Sinolight) Co., Ltd.","award":["Z02022YY01"],"award-info":[{"award-number":["Z02022YY01"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>The lactic acid bacteria Streptococcus thermophilus and Lactobacillus helveticus are commonly used as starter cultures in dairy product production. This study aimed to investigate the characteristics of fermented milk using different ratios of these strains and analyze the changes in volatile compounds during fermentation and storage. A 10:1 ratio of Streptococcus thermophilus CICC 6063 to Lactobacillus helveticus CICC 6064 showed optimal fermentation time (4.2 h), viable cell count (9.64 log10 colony-forming units\/mL), and sensory evaluation score (79.1 points). In total, 56 volatile compounds were identified and quantified by solid-phase microextraction and gas chromatography\u2013mass spectrometry (SPME-GC-MS), including aldehydes, ketones, acids, alcohols, esters, and others. Among these, according to VIP analysis, 2,3-butanedione, acetoin, 2,3-pentanedione, hexanoic acid, acetic acid, acetaldehyde, and butanoic acid were identified as discriminatory volatile metabolites for distinguishing between different time points. Throughout the fermentation and storage process, the levels of 2,3-pentanedione and acetoin exhibited synergistic dynamics. These findings enhance our understanding of the chemical and molecular characteristics of milk fermented with Streptococcus thermophilus and Lactobacillus helveticus, providing a basis for improving the flavor and odor of dairy products during fermentation and storage.<\/jats:p>","DOI":"10.3390\/molecules29061257","type":"journal-article","created":{"date-parts":[[2024,3,12]],"date-time":"2024-03-12T12:14:40Z","timestamp":1710245680000},"page":"1257","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Fermentation Characteristics of Fermented Milk with Streptococcus thermophilus CICC 6063 and Lactobacillus helveticus CICC 6064 and Volatile Compound Dynamic Profiles during Fermentation and Storage"],"prefix":"10.3390","volume":"29","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-6441-0101","authenticated-orcid":false,"given":"Xiaoxin","family":"Zhao","sequence":"first","affiliation":[{"name":"China Center of Industrial Culture Collection, China National Research Institute of Food and Fermentation Industries Co., Ltd., Beijing 100015, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanyuan","family":"Ge","sequence":"additional","affiliation":[{"name":"China Center of Industrial Culture Collection, China National Research Institute of Food and Fermentation Industries Co., Ltd., Beijing 100015, China"},{"name":"College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuejian","family":"Yu","sequence":"additional","affiliation":[{"name":"China Center of Industrial Culture Collection, China National Research Institute of Food and Fermentation Industries Co., Ltd., Beijing 100015, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chong","family":"Liu","sequence":"additional","affiliation":[{"name":"China Center of Industrial Culture Collection, China National Research Institute of Food and Fermentation Industries Co., Ltd., Beijing 100015, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haizhi","family":"Li","sequence":"additional","affiliation":[{"name":"China Center of Industrial Culture Collection, China National Research Institute of Food and Fermentation Industries Co., Ltd., Beijing 100015, China"},{"name":"College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xi","family":"Wang","sequence":"additional","affiliation":[{"name":"China Center of Industrial Culture Collection, China National Research Institute of Food and Fermentation Industries Co., Ltd., Beijing 100015, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Su","family":"Yao","sequence":"additional","affiliation":[{"name":"China Center of Industrial Culture Collection, China National Research Institute of Food and Fermentation Industries Co., Ltd., Beijing 100015, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,3,12]]},"reference":[{"key":"ref_1","unstructured":"Codex Alimentarius (2011). Milk and Milk Products, FAO\/WHO."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1002\/jib.569","article-title":"Pre-fermentation with lactic acid bacteria in sour beer production","volume":"125","author":"Dysvik","year":"2019","journal-title":"J. Inst. Brew."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1038\/s41575-020-00390-5","article-title":"The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on fermented foods","volume":"18","author":"Marco","year":"2021","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"897","DOI":"10.1007\/s00253-014-6130-8","article-title":"Putrescine production via the agmatine deiminase pathway increases the growth of Lactococcus lactis and causes the alkalinization of the culture medium","volume":"99","author":"Linares","year":"2015","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.tifs.2003.09.004","article-title":"Lactic acid bacteria as functional starter cultures for the food fermentation industry","volume":"15","author":"Leroy","year":"2004","journal-title":"Trends Food Sci. Technol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Gao, S., Jiang, Y., Zhang, X., Cui, S., Liu, X., Zhao, J., Zhang, H., and Chen, W. (2022). Comparative Peptidomics Analysis of Milk Fermented by Lactobacillus helveticus. Foods, 11.","DOI":"10.3390\/foods11233885"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Taverniti, V., and Guglielmetti, S. (2012). Health-promoting properties of Lactobacillus helveticus. Front. Microbiol., 3.","DOI":"10.3389\/fmicb.2012.00392"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Hassan, M.U., Nayab, H., Shafique, F., Williamson, M.P., Almansouri, T.S., Asim, N., Shafi, N., Attacha, S., Khalid, M., and Ali, N. (2020). Probiotic properties of Lactobacillus helveticus and Lactobacillus plantarum isolated from traditional Pakistani yoghurt. BioMed Res. Int., 2020.","DOI":"10.1155\/2020\/8889198"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1007\/s00394-004-0441-y","article-title":"The effect of Lactobacillus helveticus fermented milk on acute changes in calcium metabolism in postmenopausal women","volume":"43","author":"Narva","year":"2004","journal-title":"Eur. J. Nutr."},{"key":"ref_10","unstructured":"Rul, F. (2017). Yogurt: Microbiology, Organoleptic Properties and Probiotic Potential, CRC Press."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1007\/s12602-017-9279-5","article-title":"Comparative growth behaviour and biofunctionality of lactic acid bacteria during fermentation of soy milk and bovine milk","volume":"10","author":"Hati","year":"2018","journal-title":"Probiotics Antimicrob. Proteins"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1002\/fsn3.2016","article-title":"A weak post-acidification Lactobacillus helveticus UV mutant with improved textural properties","volume":"9","author":"Guan","year":"2021","journal-title":"Food Sci. Nutr."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1046\/j.1365-2672.2001.01219.x","article-title":"Enhanced flavour formation by combination of selected lactococci from industrial and artisanal origin with focus on completion of a metabolic pathway","volume":"90","author":"Ayad","year":"2001","journal-title":"J. Appl. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"191","DOI":"10.3168\/jds.2019-17312","article-title":"Characteristics of volatile flavor components in traditional fermented yak milk produced in different ecoregions of the Qinghai-Tibetan plateau","volume":"103","author":"Fang","year":"2020","journal-title":"J. Dairy Sci."},{"key":"ref_15","unstructured":"Krastanov, A., Yeboah, P.J., Wijemanna, N.D., Eddin, A.S., Ayivi, R.D., and Ibrahim, S.A. (2023). Current Issues and Advances in the Dairy Industry, IntechOpen."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1010","DOI":"10.1080\/10942912.2010.513024","article-title":"Volatile profile of non-fermented milk and milk fermented by Bifidobacterium animalis subsp","volume":"15","author":"Zareba","year":"2012","journal-title":"lactis. Int. J. Food Prop."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/j.foodchem.2009.04.081","article-title":"Characterisation by SPME\u2013GC\u2013MS of the volatile profile of a Spanish soft cheese PDO Torta del Casar during ripening","volume":"118","author":"Delgado","year":"2010","journal-title":"Food Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"7775","DOI":"10.1128\/AEM.01122-10","article-title":"Mixed-culture transcriptome analysis reveals the molecular basis of mixed-culture growth in Streptococcus thermophilus and Lactobacillus bulgaricus","volume":"76","author":"Sieuwerts","year":"2010","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1007\/978-3-642-37916-1_19","article-title":"Screening of the Best Strain for Naked Oat Fermentation Beverage and its Production Process Study","volume":"Volume 1","author":"Wang","year":"2014","journal-title":"Proceedings of the 2012 International Conference on Applied Biotechnology (ICAB 2012)"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1093\/chemse\/1.3.339","article-title":"Odor intensity: Mixtures and masking","volume":"1","author":"Cain","year":"1975","journal-title":"Chem. Senses"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/j.tifs.2019.03.009","article-title":"Cheese ripening: A review on modern technologies towards flavor enhancement, process acceleration and improved quality assessment","volume":"88","author":"Khattab","year":"2019","journal-title":"Trends Food Sci. Technol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"916","DOI":"10.1016\/j.foodhyd.2007.05.004","article-title":"Correlation between rheological properties and limonene release in pectin gels using an electronic nose","volume":"22","author":"Monge","year":"2008","journal-title":"Food Hydrocoll."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4374","DOI":"10.1128\/AEM.02725-08","article-title":"Fermentation pH influences the physiological-state dynamics of Lactobacillus bulgaricus CFL1 during pH-controlled culture","volume":"75","author":"Rault","year":"2009","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1693","DOI":"10.1111\/1750-3841.13779","article-title":"Effects of 4 probiotic strains in coculture with traditional starters on the flavor profile of yogurt","volume":"82","author":"Tian","year":"2017","journal-title":"J. Food Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3886","DOI":"10.1007\/s13197-013-0943-9","article-title":"Effect of carbon dioxide on sensory attributes, physico-chemical parameters and viability of Probiotic L. helveticus MTCC 5463 in fermented milk","volume":"51","author":"Shah","year":"2014","journal-title":"J. Food Sci. Technol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/j.foodhyd.2014.01.015","article-title":"Structure development and acidification kinetics in fermented milk containing oat \u03b2-glucan, a yogurt culture and a probiotic strain","volume":"39","author":"Lazaridou","year":"2014","journal-title":"Food Hydrocoll."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1002\/fbe2.12033","article-title":"Analysis of cofermentation characteristics of Lactobacillus bulgaricus and Streptococcus thermophilus based on microrheology","volume":"1","author":"Yang","year":"2022","journal-title":"Food Bioeng."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.foodres.2017.09.097","article-title":"Rennet-induced coagulation properties of yak casein micelles: A comparison with cow casein micelles","volume":"102","author":"Zhang","year":"2017","journal-title":"Food Res. Int."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Zhong, Z., Hu, R., Zhao, J., Liu, W., Kwok, L.Y., Sun, Z., Zhang, H., and Chen, Y. (2021). Acetate kinase and peptidases are associated with the proteolytic activity of Lactobacillus helveticus isolated from fermented food. Food Microbiol., 94.","DOI":"10.1016\/j.fm.2020.103651"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1007\/s13594-011-0010-0","article-title":"Acid coagulation behavior of homogenized milk: Effect of interacting and non-interacting droplets observed by rheology and diffusing wave spectroscopy","volume":"91","author":"Titapiccolo","year":"2011","journal-title":"Dairy Sci. Technol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"938","DOI":"10.1080\/10408390903044081","article-title":"Volatile flavor compounds in yogurt: A review","volume":"50","author":"Cheng","year":"2010","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Zhang, L., Mi, S., Liu, R., Sang, Y., and Wang, X. (2020). Evaluation of volatile compounds in milks fermented using traditional starter cultures and probiotics based on odor activity value and chemometric techniques. Molecules, 25.","DOI":"10.3390\/molecules25051129"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1002\/ffj.1945","article-title":"Effects of different commercial lipases on the volatile profile of lipolysed milk fat","volume":"24","author":"Wang","year":"2009","journal-title":"Flavour Fragr. J."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Cadwallader, K.R., Singh, T.K., and Jerrell, J.P. (2007). Streamlined Analysis of Short-, Medium-, and Long-Chain Free Fatty Acids in Dairy Products, ACS Publications.","DOI":"10.1021\/bk-2007-0971.ch006"},{"key":"ref_35","first-page":"25","article-title":"Nutritional value and current research status of goat milk","volume":"36","author":"Liu","year":"2013","journal-title":"J. Dairy Sci. Technol"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1016\/j.jfoodeng.2011.09.020","article-title":"A review of methods of low alcohol and alcohol-free beer production","volume":"108","author":"Silva","year":"2012","journal-title":"J. Food Eng."},{"key":"ref_37","first-page":"5","article-title":"Intensification of the synthesis of flavour compounds in yogurt by milk enrichment with their precursors","volume":"15","author":"Baranowska","year":"2006","journal-title":"Pol. J. Food Nutr. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1109","DOI":"10.1128\/AEM.71.2.1109-1113.2005","article-title":"High-level acetaldehyde production in Lactococcus lactis by metabolic engineering","volume":"71","author":"Bongers","year":"2005","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Villavicencio, J.D., Zoffoli, J.P., Plotto, A., and Contreras, C. (2021). Aroma compounds are responsible for an herbaceous off-flavor in the sweet cherry (Prunus avium L.) Cv. regina during fruit development. Agronomy, 11.","DOI":"10.3390\/agronomy11102020"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Dan, T., Ren, W., Liu, Y., Tian, J., Chen, H., Li, T., and Liu, W. (2019). Volatile flavor compounds profile and fermentation characteristics of milk fermented by Lactobacillus delbrueckii subsp. bulgaricus. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.02183"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1128","DOI":"10.3168\/jds.2011-4834","article-title":"Diacetyl levels and volatile profiles of commercial starter distillates and selected dairy foods","volume":"95","author":"Wijaya","year":"2012","journal-title":"J. Dairy Sci."},{"key":"ref_42","unstructured":"Vedamuthu, E.R. (2013). Manufacturing Yogurt and Fermented Milks, John Wiley & Sons."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Zhao, M., Li, H., Zhang, D., Li, J., Wen, R., Ma, H., Zou, T., Hou, Y., and Song, H. (2023). Variation of Aroma Components of Pasteurized Yogurt with Different Process Combination before and after Aging by DHS\/GC-O-MS. Molecules, 28.","DOI":"10.3390\/molecules28041975"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/j.ijfoodmicro.2018.08.030","article-title":"The addition of citrate stimulates the production of acetoin and diacetyl by a citrate-positive Lactobacillus crustorum strain during wheat sourdough fermentation","volume":"289","author":"Comasio","year":"2019","journal-title":"Int. J. Food Microbiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"169","DOI":"10.3168\/jds.S0022-0302(96)76348-8","article-title":"Compounds involved in the flavor of surface mold-ripened cheeses: Origins and properties","volume":"79","author":"Molimard","year":"1996","journal-title":"J. Dairy Sci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2488","DOI":"10.3168\/jds.2016-11528","article-title":"Characterization of volatile compounds in fermented milk using solid-phase microextraction methods coupled with gas chromatography-mass spectrometry","volume":"100","author":"Dan","year":"2017","journal-title":"J. Dairy Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2741","DOI":"10.1080\/10408398.2020.1858269","article-title":"Role of lactic acid bacteria in flavor development in traditional Chinese fermented foods: A review","volume":"62","author":"Hu","year":"2022","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1016\/S0958-6946(02)00124-3","article-title":"Key odorants in various cheese types as determined by gas chromatography-olfactometry","volume":"12","author":"Curioni","year":"2002","journal-title":"Int. Dairy J."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1016\/j.idairyj.2005.07.008","article-title":"Autolysis of selected Lactobacillus helveticus adjunct strains during Cheddar cheese ripening","volume":"16","author":"Kenny","year":"2006","journal-title":"Int. Dairy J."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1016\/j.idairyj.2004.08.024","article-title":"Role, mechanisms and control of lactic acid bacteria lysis in cheese","volume":"15","author":"Lortal","year":"2005","journal-title":"Int. Dairy J."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1111\/j.1541-4337.2011.00151.x","article-title":"Scientific and technical aspects of yogurt aroma and taste: A review","volume":"10","author":"Routray","year":"2011","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_52","unstructured":"(2009). Milk Products\u2014Determination of the Acidification Activity of Dairy Cultures by Continuous pH Measurement (CpH) (Standard No. ISO 26323:2009(E))."},{"key":"ref_53","unstructured":"(2016). Food Safety National Standard for the Determination of Food Acidity (Standard No. GB5009.239-2016)."},{"key":"ref_54","unstructured":"(2016). National Standard for Food Safety Food Microbiology Inspection Lactic Acid Bacteria Inspection (Standard No. GB4789.35-2016)."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.postharvbio.2017.02.008","article-title":"Quality tracing of peanuts using an array of metal-oxide based gas sensors combined with chemometrics methods","volume":"128","author":"Xu","year":"2017","journal-title":"Postharvest Biol. Technol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"5654","DOI":"10.3168\/jds.2021-21733","article-title":"Inoculum size of co-fermentative culture affects the sensory quality and volatile metabolome of fermented milk over storage","volume":"105","author":"Zhang","year":"2022","journal-title":"J. Dairy Sci."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Zeng, H., Wang, Y., Han, H., Cao, Y., and Wang, B. (2022). Changes in Key Aroma Compounds and Esterase Activity of Monascus-Fermented Cheese across a 30-Day Ripening Period. Foods, 11.","DOI":"10.3390\/foods11244026"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"105473","DOI":"10.1016\/j.idairyj.2022.105473","article-title":"Screening of mixed-species starter cultures for increasing flavour during fermentation of milk","volume":"135","author":"Dan","year":"2022","journal-title":"Int. Dairy J."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/29\/6\/1257\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:12:25Z","timestamp":1760105545000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/29\/6\/1257"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,12]]},"references-count":58,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2024,3]]}},"alternative-id":["molecules29061257"],"URL":"https:\/\/doi.org\/10.3390\/molecules29061257","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,3,12]]}}}