Investigation of new exopolysaccharides produced by strains of donkey milk
-
Arevik Israyelyan
,
Tsovinar Balabekyan
,
Lara Aleksanyan
,
Inesa Sahakyan
,
Anna Gasparyan
,
Flora Тkhruni
Abstract
Background and Objectives: It has been shown that strains of the genus Enterococcus isolated from donkey milk from different regions of the Republic of Armenia have antimicrobial activity, synthesize different polysaccharides and produce disaccharide polymers (glucose and galactose). The quantitative synthesis of polysaccharides (8-15%) depends on the composition of the nutrient medium, temperature and growing time.
Materials and Methods: Species identification of LAB strains was confirmed by 16S rDNA gene sequencing method using universal primers LAB strains. The methods used for extraction, purification and detection of exopolysaccharides are based on the method of Sørensen et al. (2022). The antibacterial activity of EPS was investigated by agar diffusion assay. Determination of the immunostimulating property was carried out using the ELISA method.
Results: The antimicrobial activity of the polysaccharide and antimicrobial protein-like fractions of the genus Enterococcus strains depends on its concentration, time of interaction with the test culture, and the species of the pathogenic bacteria.
Conclusion: The obtained results were shown that strains isolated from fermented donkey milk that are capable of synthesizing two substances of different nature with high antimicrobial properties during the growth process are promising for further research and application for their use as probiotics and biopreparations in pharmaceuticals.
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9. Wang CL, Huang TH, Liang TW, Fang CY, Wang SL. Production and characterization of exopolysaccharides and antioxidant from Paenibacillus sp. TKU023. N Biotechnol 2011; 28: 559-565.
10. Liu Z, Zhang Z, Qiu L, Zhang F, Xu X, Wei H, et al. Characterization and bioactivities of the exopolysaccharide from a probiotic strain of Lactobacillus plantarum WLPL04. J Dairy Sci 2017; 100: 6895-6905.
11. Nehal F, Sahnoun M, Smaoui S, Jaouadi B, Bejar S, Mohammed S. Characterization high production and antimicrobial activity of exopolysaccharides from Lactococcuslactis F-mou. Microb Pathog 2019; 132: 10-19.
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13. Zhou K, Zeng Y, Yang M, Chen S, He L, Ao X, et al. Production, purification and structural study of an exopolysaccharide from Lactobacillus plantarum BC-25. Carbohydr Polym 2016; 144: 205-214.
14. Zhang L, Liu C, Li D, Zhao Y, Zhang X, Zeng X, et al. Antioxidant activity of an exopolysaccharide isolated from Lactobacillus plantarum C88. Int J Biol Macromol 2013; 54: 270-275.
15. Riaz Rajoka MS, Wu Y, Mehwish HM, Bansal M, Zhao L. Lactobacillus exopolysaccharides: New perspectives on engineering strategies, physiochemical functions, and immunomodulatory effects on host health. Trends Food Sci Technol 2020; 103: 36-48.
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17. Sasikumar K, Kozhummal Vaikkath D, Devendra L. An exopolysaccharide (EPS) from a Lactobacillus plantarum BR2 with potential benefits for making functional foods. Bioresour Technol 2017; 241: 1152-1156.
18. Ishimwe N, Daliri EB, Lee BH, Fang F, Du G. The perspective on cholesterol lowering mechanisms of probiotics. Mol Nutr Food Res 2015; 59: 94-105.
19. Michael DR, Davies TS, Moss JWE, Calvente DL, Ramji DP, Marchesi JR, et al. The anti-cholesterolaemic effect of a consortium of probiotics: An acute study in C57BL/6J mice. Sci Rep 2017; 7: 2883.
20. Xiu L, Sheng S, Hu Z, Liu Y, Li J, Zhang H, et al. Exopolysaccharides from Lactobacillus kiferi as adjuvant enhanced the immuno-protective against staphylococcus aureus infection. Int J Biol Macromol 2020; 161: 10-23.
21. Werning ML, Hernández-Alcántara AM, Ruiz MJ, Soto LP, Dueñas MT, López P, et al. Biological functions of exopolysaccharides from lactic acid bacteria and their potential benefits for humans and farmed animals. Foods 2022; 11: 1284.
22. Tkhruni FN, Karapetyan KJ, Danova ST, Dimova SG, KarimpurFA. Probiotic properties of endemic strains of lactic acid bacteria. J BioSci Biotechnol 2013; 2: 109-115.
23. Sørensen HM, Rochfort KD, Maye S, MacLeod G, Brabazon D, Loscher C, et al. Exopolysaccharides of Lactic acid bacteria: Production, purification and health benefits towards functional food. Nutrients 2022; 14: 2938.
24. Bokulich NA, Amiranashvili L, Chitchyan K, Ghazanchyan N, Darbinyan K, Gagelidze N, et al. Microbial biogeography of the transnational fermented milk matsoni. Food Microbiol 2015; 50: 12-19.
25. Israyelyan AL, TkhruniFN. Investigation of the properties of endemic LAB strains of genus Enterococcus isolated from samples matsoon. J Biotechnol Biochem 2017; 3: 41-45.
26. Cavallarin L, Giribaldi M, de Los Dolores Soto-Del Rio M, Valle E, Barbarino G, Gennero MS, et al. A survey on the milk chemical and microbiological quality in dairy donkey farms located in North Western Italy. Food Control 2015; 50: 230-235.
27. Perna A, Intaglietta I, Simonetti A, Gambacorta E. Donkey milk for manufacture of novel functional fermented beverages. J Food Sci 2015; 80: S1352-S1359.
28. Tkhruni FN, Aghajanyan AE, Balabekyan TR, Khachatryan TV, Karapetyan KJ. Characteristic of bacteriocins of Lactobacillus rhamnosus BTK 20-12 potential probiotic strain. Probiotics Antimicrob Proteins 2020; 12: 716-724.
29. Parente E, Brienza C, Moles M, Ricciardi A. A comparison of methods for the measurement of bacteriocin activity. J Microbiol Methods 1995; 22: 95-108.
30. Bajpai V, Majumder R, Rather IA, Kim K. Extraction, isolation and purification of exopolysaccharide from lactic acid bacteria using ethanol precipitation method. Bangladesh J Pharmacol 2016; 11: 573-576.
31. Engvall E. The ELISA, enzyme-linked immunosorbent assay. Clin Chem 2010; 56: 319-320.
32. Israyelyan A, BalabekyanTs, Sargsyan Z, Tkhruni F. Effect of some metabiotics from strains of the genus Enterococcus on the growth of pathogenic bacteria that Cause Pneumonia. Biol Bull 2024; 51: 91-97.
2. Karapetyan K, Tkhruni F, Chitchyan К, Balabekyan Ts, Khachatryan T, Verdyan A (2020). Using of Lactic Acid Bacteria for Biopreservation of Dairy Products. International Conference on Biotechnology and Health (ICBH) pp: 84-85.
3. Afrikian E. Studies of lactic-acid bacteria in Armenia with emphasis on radioprotective properties. Environ Syst Decis 2012; 32: 256-268.
4. Shenderov BA. Metabiotics: Novel idea or natural development of probiotic conception. Microb Ecol Health Dis 2013; 24: 10.3402/mehd.v24i0.20399.
5. Tkhruni FN, Israyelyan AL, Karapetyan KR, Balabekyan Ts R, Khachatryan LM, Khachatryan TV. Comparative аntimicrobial activity of some metabiotics synthesized by lactic acid bacteria. New Armen Med J 2023; 17: 98-109.
6. Mokoena MP. Lactic acid bacteria and their bacteriocins: Classification, biosynthesis and applications against Uropathogens: A Mini-Review. Molecules 2017; 22: 1255.
7. Dertli E, Mercan E, Arici M, Yilmaz MT, Sagdic O. Characterisation of lactic acid bacteria from turkish sourdough and determination of their exopolysaccharide (EPS) production characteristics. LWT Food Sci Technol 2016; 71: 116-124.
8. Jurášková D, Ribeiro SC, Silva CCG. Exopolysaccharides produced by lactic acid bacteria: from biosynthesis to health-promoting properties. Foods 2022; 11: 156.
9. Wang CL, Huang TH, Liang TW, Fang CY, Wang SL. Production and characterization of exopolysaccharides and antioxidant from Paenibacillus sp. TKU023. N Biotechnol 2011; 28: 559-565.
10. Liu Z, Zhang Z, Qiu L, Zhang F, Xu X, Wei H, et al. Characterization and bioactivities of the exopolysaccharide from a probiotic strain of Lactobacillus plantarum WLPL04. J Dairy Sci 2017; 100: 6895-6905.
11. Nehal F, Sahnoun M, Smaoui S, Jaouadi B, Bejar S, Mohammed S. Characterization high production and antimicrobial activity of exopolysaccharides from Lactococcuslactis F-mou. Microb Pathog 2019; 132: 10-19.
12. Li W, Ji J, Rui X, Yu J, Tang W, Chen X, et al. Production of exopolysaccharides by Lactobacillus helveticus MB2-1 and its functional characteristics in vitro. LWT Food Sci Technol 2014; 59: 732-739.
13. Zhou K, Zeng Y, Yang M, Chen S, He L, Ao X, et al. Production, purification and structural study of an exopolysaccharide from Lactobacillus plantarum BC-25. Carbohydr Polym 2016; 144: 205-214.
14. Zhang L, Liu C, Li D, Zhao Y, Zhang X, Zeng X, et al. Antioxidant activity of an exopolysaccharide isolated from Lactobacillus plantarum C88. Int J Biol Macromol 2013; 54: 270-275.
15. Riaz Rajoka MS, Wu Y, Mehwish HM, Bansal M, Zhao L. Lactobacillus exopolysaccharides: New perspectives on engineering strategies, physiochemical functions, and immunomodulatory effects on host health. Trends Food Sci Technol 2020; 103: 36-48.
16. Rahbar Saadat Y, Yari Khosroushahi A, Pourghassem Gargari B. A comprehensive review of anticancer, immunomodulatory and health beneficial effects of the lactic acid bacteria exopolysaccharides. Carbohydr Polym 2019; 217: 79-89.
17. Sasikumar K, Kozhummal Vaikkath D, Devendra L. An exopolysaccharide (EPS) from a Lactobacillus plantarum BR2 with potential benefits for making functional foods. Bioresour Technol 2017; 241: 1152-1156.
18. Ishimwe N, Daliri EB, Lee BH, Fang F, Du G. The perspective on cholesterol lowering mechanisms of probiotics. Mol Nutr Food Res 2015; 59: 94-105.
19. Michael DR, Davies TS, Moss JWE, Calvente DL, Ramji DP, Marchesi JR, et al. The anti-cholesterolaemic effect of a consortium of probiotics: An acute study in C57BL/6J mice. Sci Rep 2017; 7: 2883.
20. Xiu L, Sheng S, Hu Z, Liu Y, Li J, Zhang H, et al. Exopolysaccharides from Lactobacillus kiferi as adjuvant enhanced the immuno-protective against staphylococcus aureus infection. Int J Biol Macromol 2020; 161: 10-23.
21. Werning ML, Hernández-Alcántara AM, Ruiz MJ, Soto LP, Dueñas MT, López P, et al. Biological functions of exopolysaccharides from lactic acid bacteria and their potential benefits for humans and farmed animals. Foods 2022; 11: 1284.
22. Tkhruni FN, Karapetyan KJ, Danova ST, Dimova SG, KarimpurFA. Probiotic properties of endemic strains of lactic acid bacteria. J BioSci Biotechnol 2013; 2: 109-115.
23. Sørensen HM, Rochfort KD, Maye S, MacLeod G, Brabazon D, Loscher C, et al. Exopolysaccharides of Lactic acid bacteria: Production, purification and health benefits towards functional food. Nutrients 2022; 14: 2938.
24. Bokulich NA, Amiranashvili L, Chitchyan K, Ghazanchyan N, Darbinyan K, Gagelidze N, et al. Microbial biogeography of the transnational fermented milk matsoni. Food Microbiol 2015; 50: 12-19.
25. Israyelyan AL, TkhruniFN. Investigation of the properties of endemic LAB strains of genus Enterococcus isolated from samples matsoon. J Biotechnol Biochem 2017; 3: 41-45.
26. Cavallarin L, Giribaldi M, de Los Dolores Soto-Del Rio M, Valle E, Barbarino G, Gennero MS, et al. A survey on the milk chemical and microbiological quality in dairy donkey farms located in North Western Italy. Food Control 2015; 50: 230-235.
27. Perna A, Intaglietta I, Simonetti A, Gambacorta E. Donkey milk for manufacture of novel functional fermented beverages. J Food Sci 2015; 80: S1352-S1359.
28. Tkhruni FN, Aghajanyan AE, Balabekyan TR, Khachatryan TV, Karapetyan KJ. Characteristic of bacteriocins of Lactobacillus rhamnosus BTK 20-12 potential probiotic strain. Probiotics Antimicrob Proteins 2020; 12: 716-724.
29. Parente E, Brienza C, Moles M, Ricciardi A. A comparison of methods for the measurement of bacteriocin activity. J Microbiol Methods 1995; 22: 95-108.
30. Bajpai V, Majumder R, Rather IA, Kim K. Extraction, isolation and purification of exopolysaccharide from lactic acid bacteria using ethanol precipitation method. Bangladesh J Pharmacol 2016; 11: 573-576.
31. Engvall E. The ELISA, enzyme-linked immunosorbent assay. Clin Chem 2010; 56: 319-320.
32. Israyelyan A, BalabekyanTs, Sargsyan Z, Tkhruni F. Effect of some metabiotics from strains of the genus Enterococcus on the growth of pathogenic bacteria that Cause Pneumonia. Biol Bull 2024; 51: 91-97.
| Files | ||
| Issue | Vol 17 No 1 (2025) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v17i1.17808 | |
| Keywords | ||
| Exopolysaccharides; Protein-like fractions; Interaction; Donkey milk; Antimicrobial activity | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Israyelyan A, Balabekyan T, Aleksanyan L, Sahakyan I, Gasparyan A, ТkhruniF. Investigation of new exopolysaccharides produced by strains of donkey milk. Iran J Microbiol. 2025;17(1):113-121.
