Skip to main content

Advertisement

Log in

Characterization of exopolysaccharide produced by probiotic Enterococcus durans DU1 and evaluation of its anti-biofilm activity

  • Original Paper
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

Exopolysaccharides (EPS) produced by lactic acid bacteria are complicated polymers with industrial applications. LAB were isolated, screened for EPS production, and their probiotic properties determined. The anti-biofilm activity of EPS was investigated. Safety of EPS-producing isolate was investigated and it was molecularly identified through 16S rRNA sequencing. Finally, anti-biofilm and emulsification activity of EPS was studied and it was characterized using FT-IR, TGA, 1H-NMR, DLS and HPLC. Thirteen LAB were isolated from dairy products. They showed probiotic characteristics like acid resistance (0–6.51 CFU ml−1) hydrophobicity (8–54.04%), autoaggregation (0% [t = 2 h]–99.8% [t = 24 h]) and coaggregation with food borne pathogens. Among them, Enterococcus durans DU1 had ability to produce EPS. EPS of Enterococcus durans DU1 showed antibiofilm activity against Y. enterocolitica (24.06–51.36%), S. aureus (12.33–49.6%), and B. cereus (11.66–27.16%). FT-IR showed this EPS had characteristic absorption peaks due to the presence of the pyran ring of sugars. 1H NMR showed that EPS has N-acetyl, methyl, and alkyl groups in its structure. The HPLC analysis showed that EPS is a heteropolysaccharide and consists of sucrose, glucose, and fructose. EPS showed significant thermal stability (20% weight loss) under 300 °C and zeta potential of − 18.1 mV. This EPS can be used in the food industry with no adverse effect on consumers.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Abid Y et al (2018) Production and structural characterization of exopolysaccharides from newly isolated probiotic lactic acid bacteria. Int J Biol Macromol 108:719–728

    Article  CAS  PubMed  Google Scholar 

  • Abushelaibi A, Al-Mahadin S, El-Tarabily K, Shah NP, Ayyash M (2017) Characterization of potential probiotic lactic acid bacteria isolated from camel milk. LWT-Food Sci Technol 79:316–325

    Article  CAS  Google Scholar 

  • Angmo K, Kumari A, Bhalla TC (2016) Probiotic characterization of lactic acid bacteria isolated from fermented foods and beverage of Ladakh. LWT-Food Sci Technol 66:428–435

    Article  CAS  Google Scholar 

  • Ayyash M, Abushelaibi A, Al-Mahadin S, Enan M, El-Tarabily K, Shah N (2018) In-vitro investigation into probiotic characterisation of Streptococcus and Enterococcus isolated from camel milk. LWT 87:478–487

    Article  CAS  Google Scholar 

  • Ayyash M et al (2020a) Characterization, bioactivities, and rheological properties of exopolysaccharide produced by novel probiotic Lactobacillus plantarum C70 isolated from camel milk. Int J Biol Macromol 144:938–946

    Article  CAS  PubMed  Google Scholar 

  • Ayyash M et al (2020b) Exopolysaccharide produced by potential probiotic Enterococcus faecium MS79: characterization, bioactivities and rheological properties influenced by salt and pH. LWT 131:109741

    Article  CAS  Google Scholar 

  • Benito Y, Pin C, Marin ML, Garcia ML, Selgas MD, Casas C (1997) Cell surface hydrophobicity and attachment of pathogenic and spoilage bacteria to meat surfaces. Meat Sci 45:419–425

    Article  CAS  PubMed  Google Scholar 

  • Caccamo MT, Gugliandolo C, Zammuto V, Magazù S (2020) Thermal properties of an exopolysaccharide produced by a marine thermotolerant Bacillus licheniformis by ATR-FTIR spectroscopy. Int J Biol Macromol 145:77–83

    Article  CAS  PubMed  Google Scholar 

  • Campana R, van Hemert S, Baffone W (2017) Strain-specific probiotic properties of lactic acid bacteria and their interference with human intestinal pathogens invasion. Gut Pathogens 9:1–12

    Article  Google Scholar 

  • Choi N-Y, Bae Y-M, Lee S-Y (2015) Cell surface properties and biofilm formation of pathogenic bacteria. Food Sci Biotechnol 24:2257–2264

    Article  CAS  Google Scholar 

  • Cirrincione S, Breuer Y, Mangiapane E, Mazzoli R, Pessione E (2018) ’Ropy’phenotype, exopolysaccharides and metabolism: STUDY on food isolated potential probiotics. LAB Microbiol Res 214:137–145

    Article  CAS  PubMed  Google Scholar 

  • CLSI (2017) Performance standards for antimicrobial susceptibility testing. Clinical and Laboratory Standards Institute Wayne

    Google Scholar 

  • De Man J, RogosaSharpe DME (1960) A medium for the cultivation of lactobacilli. J Appl Bacteriol 23:130–135

    Article  Google Scholar 

  • Drakou E-M, Amorim CL, Castro PM, Panagiotou F, Vyrides I (2018) Wastewater valorization by pure bacterial cultures to extracellular polymeric substances (EPS) with high emulsifying potential and flocculation activities. Waste Biomass Valoriz 9:2557–2564

    Article  CAS  Google Scholar 

  • EL-Adawi HI, Khalil MA, EL-Sheekh MM, El-Deeb NM, Hussein MZ (2012) Cytotoxicity assay and antioxidant activities of the lactic acid bacterial strains. Afr J Microbiol Res 6:1700–1712

    Google Scholar 

  • Gunyakti A, Asan-Ozusaglam M (2019) Lactobacillus gasseri from human milk with probiotic potential and some technological properties. LWT 109:261–269

    Article  CAS  Google Scholar 

  • Han M, Du C, Xu Z-Y, Qian H, Zhang W-G (2016) Rheological properties of phosphorylated exopolysaccharide produced by Sporidiobolus pararoseus JD-2. Int J Biol Macromol 88:603–613

    Article  CAS  PubMed  Google Scholar 

  • Hou Q, Bai X, Li W, Gao X, Zhang F, Sun Z, Zhang H (2018) Design of primers for evaluation of lactic acid bacteria populations in complex biological samples. Front Microbiol 9:2045

    Article  PubMed  PubMed Central  Google Scholar 

  • Imran MYM et al (2016) Statistical optimization of exopolysaccharide production by Lactobacillus plantarum NTMI05 and NTMI20. Int J Biol Macromol 93:731–745

    Article  CAS  PubMed  Google Scholar 

  • Kavitake D, Balyan S, Devi PB, Shetty PH (2020) Evaluation of oil-in-water (O/W) emulsifying properties of galactan exopolysaccharide from Weissella confusa KR780676. J Food Sci Technol 57:1579–1585

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Koebmann BJ, Nilsson D, Kuipers OP, Jensen PR (2000) The membrane-bound H+-ATPase complex is essential for growth of Lactococcus lactis. J Bacteriol 182:4738–4743

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Korcz E, Varga L (2021) Exopolysaccharides from lactic acid bacteria: Techno-functional application in the food industry. Trends Food Sci Technol 110:375–384

  • Lakra AK, Domdi L, Hanjon G, Tilwani YM, Arul V (2020) Some probiotic potential of Weissella confusa MD1 and Weissella cibaria MD2 isolated from fermented batter. LWT 125:109261

    Article  CAS  Google Scholar 

  • Lee S-H, Kim S-J, Lee Y-S, Song M-D, Kim I-H, Won H-S (2011) De novo generation of short antimicrobial peptides with simple amino acid composition. Regul Pept 166:36–41

    Article  CAS  PubMed  Google Scholar 

  • Lin X, Xia Y, Yang Y, Wang G, Zhou W, Ai L (2020a) Probiotic characteristics of Lactobacillus plantarum AR113 and its molecular mechanism of antioxidant. LWT 126:109278

    Article  CAS  Google Scholar 

  • Lin X, Xia Y, Yang Y, Wang G, Zhou W, Ai L (2020b) Probiotic characteristics of Lactobacillus plantarum AR113 and its molecular mechanism of antioxidant. LWT 126:109278

    Article  CAS  Google Scholar 

  • Liu T, Zhou K, Yin S, Liu S, Zhu Y, Yang Y, Wang C (2019) Purification and characterization of an exopolysaccharide produced by Lactobacillus plantarum HY isolated from home-made Sichuan Pickle. Int J Biol Macromol 134:516–526

    Article  CAS  PubMed  Google Scholar 

  • Mahdhi A et al (2017) Extracellular polysaccharide derived from potential probiotic strain with antioxidant and antibacterial activities as a prebiotic agent to control pathogenic bacterial biofilm formation. Microb Pathog 109:214–220

    Article  CAS  PubMed  Google Scholar 

  • Mallappa RH, Singh DK, Rokana N, Pradhan D, Batish VK, Grover S (2019) Screening and selection of probiotic Lactobacillus strains of Indian gut origin based on assessment of desired probiotic attributes combined with principal component and heatmap analysis. LWT 105:272–281

    Article  CAS  Google Scholar 

  • Matsumoto M, Ohishi H, Benno Y (2004) H+-ATPase activity in Bifidobacterium with special reference to acid tolerance. Int J Food Microbiol 93:109–113

    Article  CAS  PubMed  Google Scholar 

  • Montoro BP, Benomar N, Lavilla Lerma L, Castillo Gutiérrez S, Gálvez A, Abriouel H (2016) Fermented Aloreña table olives as a source of potential probiotic Lactobacillus pentosus strains. Front Microbiol 7:1583

    Article  PubMed  PubMed Central  Google Scholar 

  • Morth JP, Pedersen BP, Buch-Pedersen MJ, Andersen JP, Vilsen B, Palmgren MG, Nissen P (2011) A structural overview of the plasma membrane Na+, K+-ATPase and H+-ATPase ion pumps. Nat Rev Mol Cell Biol 12:60–70

    Article  CAS  PubMed  Google Scholar 

  • Mulaw G, Sisay Tessema T, Muleta D, Tesfaye A (2019) In vitro evaluation of probiotic properties of lactic acid bacteria isolated from some traditionally fermented Ethiopian food products. Int J Microbiol 2019:7179514

  • Nambiar RB, Sellamuthu PS, Perumal AB, Sadiku ER, Phiri G, Jayaramudu J (2018) Characterization of an exopolysaccharide produced by Lactobacillus plantarum HM47 isolated from human breast milk. Process Biochem 73:15–22

    Article  CAS  Google Scholar 

  • Palaniyandi SA, Damodharan K, Suh J-W, Yang SH (2018) Functional characterization of an exopolysaccharide produced by Bacillus sonorensis MJM60135 isolated from ganjang. J Microbiol Biotechnol 28:663–670

    Article  CAS  PubMed  Google Scholar 

  • Polak-Berecka M, Choma A, Waśko A, Górska S, Gamian A, Cybulska J (2015) Physicochemical characterization of exopolysaccharides produced by Lactobacillus rhamnosus on various carbon sources. Carbohydr Polym 117:501–509

    Article  CAS  PubMed  Google Scholar 

  • Rahnama Vosough P, Habibi Najafi MB, Edalatian Dovom MR, Javadmanesh A, Mayo B (2021) Evaluation of antioxidant, antibacterial and cytotoxicity activities of exopolysaccharide from Enterococcus strains isolated from traditional Iranian Kishk. J Food Meas Characte 15:5221–5230

    Article  Google Scholar 

  • Rajoka MSR et al (2019) Characterization and anti-tumor activity of exopolysaccharide produced by Lactobacillus kefiri isolated from Chinese kefir grains. J Funct Foods 63:103588

    Article  CAS  Google Scholar 

  • Saif FAA, Sakr EA (2020) Characterization and bioactivities of exopolysaccharide produced from probiotic Lactobacillus plantarum 47FE and Lactobacillus pentosus 68FE. Bioactive Carbohydr Diet Fibre 24:100231

    Article  CAS  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees Mol Biol Evol 4:406–425

  • Sarikaya H, Aslim B, Yuksekdag Z (2017) Assessment of anti-biofilm activity and bifidogenic growth stimulator (BGS) effect of lyophilized exopolysaccharides (l-EPSs) from Lactobacilli strains. Int J Food Prop 20:362–371

    Article  CAS  Google Scholar 

  • Sasikumar K, Vaikkath DK, Devendra L, Nampoothiri KM (2017) An exopolysaccharide (EPS) from a Lactobacillus plantarum BR2 with potential benefits for making functional foods. Bioresour Technol 241:1152–1156

    Article  CAS  PubMed  Google Scholar 

  • Sayem S et al (2011) Anti-biofilm activity of an exopolysaccharide from a sponge-associated strain of Bacillus licheniformis. Microb Cell Fact 10:1–12

    Article  CAS  Google Scholar 

  • Sharma V, Harjai K, Shukla G (2018) Effect of bacteriocin and exopolysaccharides isolated from probiotic on P. aeruginosa PAO1 biofilm. Folia Microbiol 63:181–190

    Article  CAS  Google Scholar 

  • Silva LA, Neto JHPL, Cardarelli HR (2019) Exopolysaccharides produced by Lactobacillus plantarum: technological properties, biological activity, and potential application in the food industry. Ann Microbiol 69:321–328

    Article  CAS  Google Scholar 

  • Singh SP, Qureshi A, Hassan W (2021) Mechanisms of action by antimicrobial agents: a review. McGill J Med 19(4):1–10

  • Smetanková J, Hladíková Z, Valach F, Zimanová M, Kohajdová Z, Greif G, Greifová M (2012) Influence of aerobic and anaerobic conditions on the growth and metabolism of selected strains of Lactobacillus plantarum. Acta Chim Slov 5:204

    Article  Google Scholar 

  • Song Y et al. (2020) Antibiofilm activity of Lactobacillus plantarum 12 exopolysaccharides against Shigella flexneri. Appl Environ Microbiol 86(15):e00694-20

  • Trabelsi L, M’sakni NH, Ouada HB, Bacha H, Roudesli S (2009) Partial characterization of extracellular polysaccharides produced by cyanobacterium Arthrospira platensis. Biotechnol Bioprocess Eng 14:27–31

    Article  CAS  Google Scholar 

  • Vettori MHPB, Franchetti SMM, Contiero J (2012) Structural characterization of a new dextran with a low degree of branching produced by Leuconostoc mesenteroides FT045B dextransucrase. Carbohyd Polym 88:1440–1444

    Article  CAS  Google Scholar 

  • Vlková E, Rada V, Popelářová P, Trojanová I, Killer J (2006) Antimicrobial susceptibility of bifidobacteria isolated from gastrointestinal tract of calves. Livest Sci 105:253–259

    Article  Google Scholar 

  • Wang J, Zhao X, Yang Y, Zhao A, Yang Z (2015) Characterization and bioactivities of an exopolysaccharide produced by Lactobacillus plantarum YW32. Int J Biol Macromol 74:119–126

    Article  CAS  PubMed  Google Scholar 

  • Wang X, Shao C, Liu L, Guo X, Xu Y, Lü X (2017) Optimization, partial characterization and antioxidant activity of an exopolysaccharide from Lactobacillus plantarum KX041. Int J Biol Macromol 103:1173–1184

    Article  CAS  PubMed  Google Scholar 

  • Won S-M, Chen S, Park KW, Yoon J-H (2020) Isolation of lactic acid bacteria from kimchi and screening of Lactobacillus sakei ADM14 with anti-adipogenic effect and potential probiotic properties. LWT 126:109296

    Article  CAS  Google Scholar 

  • Wu W, Pang B, Yang R, Liu G, Ai C, Jiang C, Shi J (2020) Improvement of the probiotic potential and yield of Lactobacillus rhamnosus cells using corn steep liquor. LWT 131:109862

    Article  CAS  Google Scholar 

  • Xiaomei G, Houming W, Guirong M (2004) Isolation, purification and structural elucidation of EPS-II, an extracellular polysaccharide from Enterococcus durans. Chem J Chin Univ 25:1288–1290

    Google Scholar 

  • Xu X et al (2020) Antibacterial potential of a novel Lactobacillus casei strain isolated from Chinese northeast sauerkraut and the antibiofilm activity of its exopolysaccharides. Food Funct 11:4697–4706

    Article  CAS  PubMed  Google Scholar 

  • Yang Z, Li S, Zhang X, Zeng X, Li D, Zhao Y, Zhang J (2010) Capsular and slime-polysaccharide production by Lactobacillus rhamnosus JAAS8 isolated from Chinese sauerkraut: potential application in fermented milk products. J Biosci Bioeng 110:53–57

    Article  CAS  PubMed  Google Scholar 

  • Ye G, Chen Y, Wang C, Yang R, Bin X (2018) Purification and characterization of exopolysaccharide produced by Weissella cibaria YB-1 from pickle Chinese cabbage. Int J Biol Macromol 120:1315–1321

    Article  CAS  PubMed  Google Scholar 

  • Zhang G, Zhang W, Sun L, Sadiq FA, Yang Y, Gao J, Sang Y (2019) Preparation screening, production optimization and characterization of exopolysaccharides produced by Lactobacillus sanfranciscensis Ls-1001 isolated from Chinese traditional sourdough. Int J Biol Macromol 139:1295–1303

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Damghan University for its financial support of this research.

Funding

No funding was secured for this manuscript.

Author information

Authors and Affiliations

Authors

Contributions

OS: investigation, methodology. FS: methodology, validation, investigation, formal analysis, data curation, writing—review & editing, supervision, project administration. AR: editing.

Corresponding author

Correspondence to Fatemeh Salimi.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

Ethics approval

Not applicable.

Consent to participate

The authors have contributed substantially to the manuscript and approved the final submission.

Consent for publication

The authors have approved the publication rules.

Additional information

Communicated by Erko Stackebrandt.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Soliemani, O., Salimi, F. & Rezaei, A. Characterization of exopolysaccharide produced by probiotic Enterococcus durans DU1 and evaluation of its anti-biofilm activity. Arch Microbiol 204, 419 (2022). https://doi.org/10.1007/s00203-022-02965-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00203-022-02965-z

Keywords

Navigation