Biotechnology production of medium for cultivation and lyophilization of lactic acid bacteria

S.O. Gujvinska, A.P. Paliy, O.V. Dunaeva, A.P. Paliy, N.V. Berezhna


The aim of our work was to develop a medium for the cultivation of lactic acid bacteria and to select the optimal protective mediums for the freeze-drying of lactic acid bacteria. The objects of research were cultures of microorganisms Lactobacillus plantarum 7 and Lactobacillus casei 27 isolated, selected and stored in the National Scientific Center Institute of Experimental and Clinical Veterinary Medicine (NSC "IECVM", Kharkiv). Cultivation of lactobacilli was carried out on medium MRS and Blaurock for 24-72 hours at the temperature of 37 °C. To develop nutrient media for the cultivation of lactobacilli, the selected components in the main composition of the medium: lactose, sodium phosphate disubstituted, sodium citrate. As components of the protective medium, solutions of sucrose, gelatinase, sodium acetate, and aerosol® were used. Because of the studies, the optimal composition of the nutrient medium was determined (lactose 1.2 %, sodium phosphate disubstituted 0.3 %, and sodium citrate 0.6 %) for the cultivation of lactic acid bacteria on which bacteria grow and actively accumulate a significant number of viable cells from 7.1 to 8.2 × 106 CFU/cm3. In the various cultivation regimes of lactobacilli, pH 7.0 is optimal, and the temperature of the inoculum incubation is 37±0,5 °C. An alternative protective medium is added with the addition of 10 % sucrose, 1.0 % gelatinase and 3.0 % aerosil, which ensures the viability of viable cells after lyophilization of strains of L. plantarum 7 and L. casei 27, respectively, 96 % and 97 %.


lactobacillus;cultivation;pH;temperature;culture medium;protective medium

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Arkad'eva, Z.A., Karryeva, D.A., Baranova, N.A. (1983). Preservation of lactic acid bacteria. Nauchnye Doki Vyss Shkoly Biol Nauki, (2),101-4. PMID: 6404311

Bannikova, L.O. (1975). Selection of lactic acid bacteria and their use in the dairy industry. Moscow: Food industry. (In Russian)

Basjul, O.V., Jamborko, G.V., Ivanycja, V.O. (2014). Influence of the composition of protective environments on preservation of viability and biotechnological properties of lyophilized bacteria Lactobacillus Plantarum ONU315. Bulletin of Odessa National University, 19 (1(34)), 9-15. (In Ukrainian)

Belletti, N., Gatti, M., Bottari, B., Neviani, E., Tabanelli, G., Gardini, F. (2009). Antibiotic resistance of Lactobacilli isolated from two Italian hard cheeses. J. Food Prot., – 72 (10), 2162-2169. PMID: 19833040

Bujalance, C., Jiménez-Valera, M., Moreno, E., Ruiz-Bravo, A. (2006). A selective differential medium for Lactobacillus plantarum. J. Microbiol. Methods, 66 (3), 572-575. doi: 10.1016/j.mimet.2006.02.005

De Man, J.C., Rogosa, M., Sharpe, M.E. (1960). A medium for the cultivation of Lactobacilli. J. Appl. Microbiol., 23 (1), 130-135. doi: 10.1111/j.1365-2672.1960.tb00188.x

Gujvinska, S.O., Paliy, A.P. (2018). Determination of antagonistic and adhesive properties of Lactobacterium and Bifidobacterium. Mikrobiol. Z., 80 (1), 36-44. doi:

Gujvinska, S.O. (2014). Selection of lactic acid bacteria for the manufacture of probiotic drugs. Veterinary medicine, 99, 196-200. (In Ukrainian)

Charteris, W.P., Kelly, P.M., Morelli, L., Collins, J.K. (2000). Effect of conjugated bile salts on antibiotic susceptibility of bile salt-tolerant Lactobacillus and Bifidobacterium isolates. J. Food Prot., 63 (10), 1369-1376. PMID: 11041136

Del, Re.B., Sgorbati, B., Miglioli, M., Palenzona, D. (2000). Adhesion, autoaggregation and hydrophobicity of 13 strains of Bifidobacterium longum. Lett. Appl. Microbiol., 31 (6), 438-442. PMID: 11123552

Jamalifar, H., Rahimi, H., Samadi, N., Shahverdi, A.R., Sharifian, Z., Hosseini, F., Eslahi, H., Fazeli, M.R. (2011). Antimicrobial activity of different Lactobacillus species against multi-drug resistant clinical isolates of Pseudomonas aeruginosa. Iran. J. Microbiol., 3(1), 21-25. PMID: 22347578

Jankowski, D.S. (2007). Selection of lactobacilli strains with promising biotic properties. Bulletin of Agrarian Science, 12, 60-63. (In Ukrainian)

Kapraljants, L.V., Liventsova, O., Tregub, N.S. (2016). Cultivation of bifido- and lactobacilli in sodium environments with selenium. Food Science and Technology, 10 (1), 26-30. (In Russian)

Khilko, T.V., Osadcha, A.I. (2004). Growth characteristics of Lactobacillus plantarum, Bacillus subtilis and Bacillus licheniformis in deep cultivation in monoculture and in conditions of coexistence. Mikrobiol. Z., 66(2), 63-68. (In Ukrainian)

Kos, B., Suskovic, J., Vukovic, S., Simpraga, M., Frece, J., Matosic, S. (2003). Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. J. Appl. Microbiol., 94 (6), 981-987. PMID: 12752805

Kovalenko, N.K. (2002). Biotechnology of cultivation of lactic acid bacteria. Dairy industry, 2, 24-25. (In Ukrainian)

Kvasnikov, E.P., Nesterenko, O.A. (1975). Lactic Acid Bacteria and Ways of Their Use. Moscow: Nauka. (In Russian)

Labinskaya, A.S. (1978). Microbiology with the technique of microbiological research. Moscow: Medicine. (In Russian)

Lahteinen, T., Malinen, E., Koort, J.M., Mertaniemi-Hannus, U., Hankimo, T., Karikoski, N., Pakkanen, S., Laine, H., Sillanpaa, H., Soderholm, H., Palva, A. (2010). Probiotic properties of Lactobacillus isolates originating from porcine intestine and feces. Anaerobe, 16 (3), 293-300. doi:

Lee, H.M., Lee, Y. (2008). A differential medium for lactic acid-producing bacteria in a mixed culture. Let. in Appl. Microbiol., 46 (6), 676-681. doi: 10.1111/j.1472-765X.2008.02371.x

Miele, E., Pascarella, F., Giannetti, E., Quaglietta, L., Baldassano, R.N., Staiano, A. (2009). Effect of a probiotic preparation (VSL#3) on induction and maintenance of remission in children with ulcerative colitis. Am. J. Gastroenterol., 104, 437-443. doi: 10.1038/ajg.2008.118

Ng, S.C., Hart, A.L., Kamm, M.A., Stagg, A.J., Knight, S.C. (2009) Mechanisms of action of probiotics: recent advances. Inflamm. Bowel Dis., 15 (2), 300-310. doi:

Patel, A.K., Ahire, J.J., Pawar, S.P., Chaudhari B.L., Chincholkar, S.B. (2009). Comparative accounts of probiotic characteristics of Bacillus spp. isolated from food wastes. Food Res. Internat., 42 (4), 505-510. doi:

Prabhurajeshwar, C., Chandrakanth, R.K. (2017). Probiotic potential of Lactobacilli with antagonistic activity against pathogenic strains: An in vitro validation for the production of inhibitory substances. Biomedical J., 40 (5), 270-283. doi:

Reid, G. (2006). Safe and efficacious probiotics: what are they? Trends Microbiol., 14 (8); 348-352. doi: 10.1016/j.tim.2006.06.006

Romanchuk, I.O. (1999). Increased survival of bacterial cultures in the production of lyophilized preparations. Bulletin of Agrarian Science, 11, 63-65. (In Ukrainian)

Schillinger, U., Holzapfel, W.H. (2003). Chapter 8 Culture media for lactic acid bacteria. Progress in Industrial Microbiology, 37, 127-140. doi:

Stiles, M.E. (1996). Biopreservation by lactic acid bacteria. Antonie Van Leeuwenhoek, 70 (2-4), 331-345. PMID: 8879414

Stoyanova, L.G., Arkadyeva, Z.A. (2000). Comparison of storage methods for lactic acid bacteria. Microbiology, 69 (1), 98-104. (In Ukrainian)

Vankerckhoven, V., Huys, G., Vancanneyt, M., Vael, C., Klare, I., Romond, M.B., et al. (2008). Biosafety assessment of probiotics used for human consumption: recommendations from the EU-PROSAFE project. Trends Food Sci. Technol., 19 (2), 102-114. doi:

Yamborko, G.V., Solovyova, I.L. (2007). The effectiveness of different ways of storing industrial strains of bacteria of the genus Lactobacillus. Microbiology and Biotechnology, 1, 53-59. (In Ukrainian)


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