Journal of food science and technology(Iran)

Journal of food science and technology(Iran)

Evaluation of the Technological Characteristics of a Lacticaseibacillus casei ANNC 26 Strain Isolated from Traditional Khiki Cheese as a Co-Starter Culture

Document Type : Original Article

Authors
Alireza Vasiee 1
Behrooz Alizadeh Behbahani 2
Parisa Ghasemi 2
Zeinab Mousavi 3
1 Department of Food Science and Technology, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
2 Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani
3 Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani,
10.48311/fsct.2026.118719.83022
Abstract
The functional efficacy of starter cultures in industrial settings relies heavily on their resilience to stress. This study comprehensively evaluated the key technological properties of an indigenous Lacticaseibacillus casei ANNC 26 strain isolated from traditional Khiki cheese; thermal resistance, exopolysaccharide (EPS) production, acidifying ability, and enzymatic activities. The thermal resistance assessment showed the strain possessed significant stability against heat stress. The highest survival (57.42%) was recorded at 50 °C, and even at 80 °C, a 25.73% survival was observed, indicating its resilience under thermal conditions. The strain showed significant EPS synthesis, induced most strongly by sucrose and glucose, validating its potential as a multifunctional bio-additive for enhancing texture and water retention. With an acidifying activity (ΔpH24h=1.48), the strain was classified as a Moderate Acidifier, making it ideal for use as an Adjunct Culture in long-ripened cheeses requiring gradual pH reduction. Furthermore, it demonstrated high proteolytic activity (21.69 ± 0.09 mm halo diameter) and "good" autolytic activity. These high enzymatic activities confirm its potential to release flavor precursors and bioactive peptides. Collectively, these favorable characteristics confirm the indigenous L. casei ANNC 26 strain as a promising Adjunct Starter candidate for significantly enhancing the texture, flavor, and stability of Khiki cheese, thereby supporting its industrial development.
Keywords
Traditional Khiki Cheese
Exopolysaccharide
Acidifying Ability
L. casei
Subjects
[1] Marco, M. L., Sanders, M. E., Gänzle, M., Arrieta, M. C., Cotter, P. D., De Vuyst, L., Hill, C., Holzapfel, W., Lebeer, S., Merenstein, D., Reid, G., Wolfe, B. E., & Hutkins, R. (2017). The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on fermented foods. Nature Reviews Gastroenterology & Hepatology, 14(3), 196–208.
[2] Rezac, S., Kok, C. R., Heermann, M., & Hutkins, R. (2018). Fermented foods as a dietary source of live organisms. Frontiers in Microbiology, 9, 1785.
[3] Liu, W., Zhang, R., Shuai, Y., Li, B., Wang, J., & He, X. (2019). Role of lactic acid bacteria in fermented foods and their health benefits. Food Science and Human Wellness, 8(2), 103–109.
[4] Tamang, J. P., Cotter, P. D., Endo, A., Han, N. S., Kort, R., Liu, S. Q., Mayo, B., Westerik, N., & Hutkins, R. (2020). Fermented foods in a global age: East meets West. Comprehensive Reviews in Food Science and Food Safety, 19(1), 184–217.
[5] LeBlanc, J. G., Milani, C., de Giori, G. S., Sesma, F., van Sinderen, D., & Ventura, M. (2017). Bacteria as vitamin suppliers to their host: A gut microbiota perspective. Current Opinion in Biotechnology, 44, 16–23.
[6] Hill, C., Guarner, F., Reid, G., Gibson, G. R., Merenstein, D. J., Pot, B., Morelli, L., Canani, R. B., Flint, H. J., Salminen, S., Calder, P. C., & Sanders, M. E. (2014). Expert consensus document: The International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nature Reviews Gastroenterology & Hepatology, 11(8), 506–514.
[7] Hayaloglu, A. A. (2014). Cheese varieties ripened under brine: A review. Trends in Food Science & Technology, 41(2), 226–259.
[8] Hayaloglu, A. A., & Farkye, N. Y. (2011). Cheese with brine: White brined cheeses. In J. W. Fuquay (Ed.), Encyclopedia of Dairy Sciences (2nd ed., pp. 783–789). Academic Press.
[9] Randazzo, C. L., Todaro, A., Pino, A., Pitino, I., Corona, O., Caggia, C., & Di Bella, S. (2018). Microbiota and metabolome during controlled and spontaneous fermentation of Nocellara Etnea table olives. Food Microbiology, 73, 136–146.
[10] Zago, M., Fornasari, M. E., Carminati, D., Burns, P., Suàrez, V., Vinderola, G., Reinheimer, J., & Giraffa, G. (2011). Characterization and probiotic potential of Lactobacillus plantarum strains isolated from cheeses. Food Microbiology, 28(5), 1033–1040.
[11] Linares, D. M., Gómez, C., Renes, E., Fresno, J. M., Tornadijo, M. E., Ross, R. P., & Stanton, C. (2017). Lactic acid bacteria and bifidobacteria with potential to design natural biofunctional health-promoting dairy foods. Frontiers in Microbiology, 8, 846.
[12] Alizadeh Behbahani, B., & Noshad, M. (2021). Isolation and Identification of Lactobacillus Strains from Behbahan Local Cheeses and Investigation of Technological and Antimicrobial Properties of These Strains against Major Food Pathogens. Iranian Journal of Nutrition Sciences and Food Technology, 16(1), 133-142.
[13] Dimitrellou, D., Kandylis, P., & Kourkoutas, Y. (2019). Thermal tolerance of probiotic microorganisms in functional dairy products: A review. Foods, 8(10), 431.
[14] García-Cano, I., Rocha-Mendoza, D., Ortega-Anaya, J., Wang, K., Kosmerl, E., & Jiménez-Flores, R. (2019). Lactic acid bacteria isolated from dairy products as potential producers of EPS and their effect on textural properties of fresh cheese. Food Microbiology, 84, 103247.
[15] Kulkarni, S., Haq, S. F., Samant, S., & Sukumaran, S. (2018). Adaptation of Lactobacillus acidophilus to thermal stress yields a thermotolerant variant which also exhibits improved survival at pH 2. Probiotics and antimicrobial proteins, 10(4), 717-727.
[16] Ogunremi, O. R., Leischtfeld, S. F., & Schwenninger, S. M. (2022). MALDI-TOF MS profiling and exopolysaccharide production properties of lactic acid bacteria from Kunu-zaki-A cereal-based Nigerian fermented beverage. International Journal of Food Microbiology, 366, 109563.
[17] Xanthopoulos, V., Hatzikamari, M., Adamidis, T., Tsakalidou, E., Tzanetakis, N., & Litopoulou‐Tzanetaki, E. (2000). Heterogeneity of Lactobacillus plantarum isolates from Feta cheese throughout ripening. Journal of Applied Microbiology, 88(6), 1056-1064.
[18] Saci, A., Gharbi, S., Djadouni, F., & Karkachi, N. (2025). Evaluation of selected functional traits in lactic acid bacteria isolated from Algerian Makatia goat milk for food applications. Acta Biologica Slovenica, 68(4).
[19] Davati, N., Tabatabaee, Y. F., Zibaee, S., Shahidi, F., & Edalatian, M. R. (2016). Isolation and identification of Lactobacillus bacteria from raw milk of Iranian one humped camel (Camelus dromedarius) and evaluation of their technological properties.
[20] Ferrando, V., Quiberoni, A., Reinhemer, J., & Suárez, V. (2015). Resistance of functional Lactobacillus plantarum strains against food stress conditions. Food microbiology, 48, 63-71.
[21] Wang, Y., Hao, F., Lu, W., Suo, X., Bellenger, E., Fu, N., ... & Chen, X. D. (2020). Enhanced thermal stability of lactic acid bacteria during spray drying by intracellular accumulation of calcium. Journal of Food Engineering, 279, 109975.
[22] Gouesbet, G., Jan, G., & Boyaval, P. (2001). Lactobacillus delbrueckii ssp. bulgaricus thermotolerance. Le Lait, 81(1-2), 301-309.
[23] Chen, X., Liu, Y., Fan, M., Wang, Z., Wu, W., & Wang, J. (2017). Thermal and chemical inactivation of Lactobacillus virulent bacteriophage. Journal of Dairy Science, 100(9), 7041-7050.
[24] Noshad, M., Alizadeh Behbahani, B., & Hojjati, M. (2021). Investigation of probiotic and technological characteristics of lactic acid bacteria isolated from native Doogh of Behbahan. Food Research Journal, 31(4), 169-186.
[25] Jurášková, D., Ribeiro, S. C., & Silva, C. C. (2022). Exopolysaccharides produced by lactic acid bacteria: from biosynthesis to health-promoting properties. Foods, 11(2), 156.
[26] Santal, A. R., Singh, N. P., & Singha, T. K. (2019). Characterization of extracellular polymeric substance producing isolates from wastewaters and their antibacterial prospective. J. Appl. Biol. Biotech, 7, 56-62.
[27] Bacosa, H. P., Kamalanathan, M., Chiu, M. H., Tsai, S. M., Sun, L., Labonté, J. M., ... & Quigg, A. (2018). Extracellular polymeric substances (EPS) producing and oil degrading bacteria isolated from the northern Gulf of Mexico. PLoS One, 13(12), e0208406.
[28] Minari, G. D., Piazza, R. D., Sass, D. C., & Contiero, J. (2024). EPS production by Lacticaseibacillus casei using glycerol, glucose, and molasses as carbon sources. Microorganisms, 12(6), 1159.
[29] Cerning, J. C. M. C., Renard, C. M. G. C., Thibault, J. F., Bouillanne, C., Landon, M., Desmazeaud, M., & Topisirovic, L. (1994). Carbon source requirements for exopolysaccharide production by Lactobacillus casei CG11 and partial structure analysis of the polymer. Applied and Environmental Microbiology, 60(11), 3914-3919.
[30] Coda, R., Di Cagno, R., Gobbetti, M., & Rizzello, C. G. (2014). Sourdough lactic acid bacteria: Exploration of non-wheat cereal-based fermentation. Food microbiology, 37, 51-58.
[31] Jialiang, Z., Huaxi, Y., Xianjun, L., Wen, Z., & Xiaodong, X. (2015). Production of exopolysaccharides by Lactobacillus fermentum F6 isolated from sourdough and its application in gluten-free bread. Food Hydrocolloids, 43, 182–188.
[32] Paul, D., Kumari, P. K., & Siddiqui, N. (2023). Yeast carotenoids: Cost-effective fermentation strategies for health care applications. Fermentation, 9(2), 147.
[33] Lin, P. C., Wu, D. T., Xie, J., Zhao, J., & Li, S. P. (2015). Characterization and comparison of bioactive polysaccharides from the tubers of Gymnadenia conopsea. Food Hydrocolloids, 43, 199-206.
[34] Rahmati, F. (2017). Characterization of Lactobacillus, Bacillus and Saccharomyces isolated from Iranian traditional dairy products for potential sources of starter cultures. AIMS microbiology, 3(4), 815.
[35] Mohammed, S., & Çon, A. H. (2021). Isolation and characterization of potential probiotic lactic acid bacteria from traditional cheese. Lwt, 152, 112319.
[36] Iqbal, M. W., Mu, W., Khan, I. M., Mohsin, A., Rehman, A., & Koko, M. Y. F. (2017). Development of probiotic soft cheese with Lactobacillus casei as adjunct culture. Journal of Academia and Industrial Research (JAIR), 6(1), 1.
[37] Barzegar, H., Alizadeh Behbahani, B., & Falah, F. (2021). Safety, probiotic properties, antimicrobial activity, and technological performance of Lactobacillus strains isolated from Iranian raw milk cheeses. Food Science & Nutrition, 9(8), 4094-4107.
[38] Hawaz, E., Guesh, T., Kebede, A., & Menkir, S. (2016). Characterization of lactic acid bacteria from camel milk and their technological properties to use as a starter culture. East African Journal of Sciences, 10(1), 49-60.
[39] Vuillemard, J. C., Amiot, J., & Gauthier, S. (1986). Evaluation de l’activité protéolytique de bactéries lactiques par une méthode de diffusion sur plaque. Microbiol. Alim. Nutr, 3, 327-332.
[40] Namdari, A., & Nejati, F. (2016). Development of Antioxidant Activity during Milk Fermentation by Wild Isolates of Lactobacillus helveticus. Applied Food Biotechnology, 3(3), 178–186.
[41] El-Salam, M. H. A., & El-Shibiny, S. (2017). Bioactive peptides of buffalo, camel, goat, sheep, mare, and yak milks and milk products. Food Reviews International, 33(1), 1–29.
[42] Fox, P. F., Guinee, T. P., Cogan, T. M., & McSweeney, P. L. H. (2017). Fundamentals of Cheese Science (2nd ed.). Springer.
[43] Ong, L., Dagastine, R. R., Kentish, S. E., & Gras, S. L. (2011). Microstructure and composition of full fat Cheddar cheese made with ultrafiltered milk retentate. Food Research International, 44(1), 91–98.
[44] Xu, Y., Wang, T., Kong, J., & Wang, H. L. (2015). Identification and functional characterization of AclB, a novel cell-separating enzyme from Lactobacillus casei. International Journal of Food Microbiology, 203, 93-100.
[45] Dalca, S., Simsek, Ö., Gursoy, O., & Yilmaz, Y. (2018). Selection of autolytic lactic acid bacteria as potential adjunct cultures to accelerate ripening of white-brined cheeses. Mljekarstvo, 68(4).