Structural characterization, biofunctionality, and in vitro digestibility of exopolysaccharides from Lactiplantibacillus plantarum E1K2R2

Nadia Aliouche , Mohamed Sifour , Selay Tornaci , Ebru Toksoy Oner , Enes Dertli , Houria Ouled-Haddar

Systems Microbiology and Biomanufacturing ›› 2026, Vol. 6 ›› Issue (1) : 4

PDF
Systems Microbiology and Biomanufacturing ›› 2026, Vol. 6 ›› Issue (1) :4 DOI: 10.1007/s43393-025-00402-y
Original Article
research-article

Structural characterization, biofunctionality, and in vitro digestibility of exopolysaccharides from Lactiplantibacillus plantarum E1K2R2

Author information +
History +
PDF

Abstract

Recently, microbial polysaccharides have gained significant importance as biomaterials, owing to their health benefits and inherent biocompatibility. Accordingly, the present study focuses on characterizing exopolysaccharides (EPS) from Lactiplantibacillus plantarum E1K2R2, investigating their structure, biocompatibility, antioxidant properties, antitumor effects, and digestibility under simulated saliva and gastrointestinal conditions. The EPS was identified as a heteropolysaccharide primarily composed of galactose and glucose monomers, with a molecular weight ranging from 3.15 × 103 to 3.95 × 105 Da. The measured particle size was 498.7 nm, while its negative zeta potential (− 7.20 mV) pointed to an acidic profile. FTIR analysis further supported this finding by identifying characteristic hydroxyl and carboxyl functional groups at 3273.57 cm−1 and 1646.91 cm−1, respectively. 1H NMR spectroscopy demonstrated that monomers in the EPS were linked by α- glycosidic bonds. Functionally, the EPS displayed dose-dependent antioxidant activity within 0.1–1 mg/mL, equivalent to 0.07–0.17 mM/mL ascorbic acid, with ABTS·+ radical scavenging ranging from 33.63 to 65.63%, demonstrating strong free radical–scavenging potential. In cytotoxicity assays, the EPS moderately inhibited HepG2 cell proliferation (6.49–20.53%) while maintaining 95% viability in normal HUVEC cells at 1 mg/mL after 24 h of incubation, indicating selective cytotoxicity toward cancer cells and good biocompatibility with normal cells. The EPS also resists simulated saliva and gastrointestinal conditions and promotes the growth of some potential probiotic lactic acid bacteria, indicating that digestive conditions do not adversely affect the EPS. These findings suggest that EPS has promising applications due to its antioxidant, antitumor, and prebiotic contexts, supported by its high biocompatibility.

Keywords

Antioxidant activity / Anti-tumor activity / Biocompatibility / Exopolysaccharides / Lactiplantibacillus plantarum E1K2R2 / Structural characterization

Cite this article

Download citation ▾
Nadia Aliouche, Mohamed Sifour, Selay Tornaci, Ebru Toksoy Oner, Enes Dertli, Houria Ouled-Haddar. Structural characterization, biofunctionality, and in vitro digestibility of exopolysaccharides from Lactiplantibacillus plantarum E1K2R2. Systems Microbiology and Biomanufacturing, 2026, 6(1): 4 DOI:10.1007/s43393-025-00402-y

登录浏览全文

4963

注册一个新账户 忘记密码

References

[1]

Andrew M, Jayaraman G. Molecular characterization and biocompatibility of exopolysaccharide produced by moderately halophilic bacterium Virgibacillus dokdonensis from the saltern of Kumta Coast. Polymers, 2022, 14(19): 3986

[2]

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

[3]

Liu Z, Zhang Z, Qiu L, Zhang F, Xu X, Wei H, Tao X. Characterization and bioactivities of the exopolysaccharide from a probiotic strain of Lactobacillus plantarum WLPL04. J Dairy Sci, 2017, 100(96895-6905

[4]

Wu J, Yan D, Liu Y, Luo X, Li Y, Cao C, Zhang L. Purification, structural characteristics, and biological activities of exopolysaccharide isolated from Leuconostoc mesenteroides SN-8. Front Microbiol, 2021, 12: 644226

[5]

Kowsalya M, Velmurugan T, Mythili R, Kim W, Sudha KG, Ali S, Rajeshkumar MP. Extraction and characterization of exopolysaccharides from Lactiplantibacillus plantarum strain PRK7 and PRK 11, and evaluation of their antioxidant, emulsion, and antibiofilm activities. Int J Biol Macromol, 2023, 242: 124842

[6]

Taylan O, Yilmaz MT, Dertli E. Partial characterization of a levan type exopolysaccharide (EPS) produced by Leuconostoc mesenteroides showing immunostimulatory and antioxidant activities. Int J Biol Macromol, 2019, 136: 436-444

[7]

Abid Y, Casillo A, Gharsallah H, Joulak I, Lanzetta R, Corsaro MM, Azabou S. Production and structural characterization of exopolysaccharides from newly isolated probiotic lactic acid bacteria. Int J Biol Macromol, 2018, 108: 719-728

[8]

Bamigbade G, Ali AH, Subhash A, Tamiello-Rosa C, Al Qudsi FR, Esposito G, Ayyash M. Structural characterization, biofunctionality, and environmental factors impacting rheological properties of exopolysaccharide produced by probiotic Lactococcus lactis C15. Sci Rep, 2023, 13(1): 17888

[9]

Wu J, Wu Z, Pan Y, Luo D, Zhong Q. Effects of different stress conditions on the production, bioactivities, physicochemical and structural characteristics of exopolysaccharides synthetized by Schleiferilactobacillus harbinensis Z171. Int J Biol Macromol, 2024, 257: 128675

[10]

Zhao D, Jiang J, Liu L, Wang S, Ping W, Ge J. Characterization of exopolysaccharides produced by Weissella confusa XG-3 and their potential biotechnological applications. Int J Biol Macromol, 2021, 178: 306-315

[11]

Kumar K, Rajulapati V, Goyal A. In vitro prebiotic potential, digestibility and biocompatibility properties of laminari-oligosaccharides produced from curdlan by β-1, 3-endoglucanase from Clostridium thermocellum. 3 Biotech, 2020, 10: 1-10

[12]

Bachtarzi N, Gomri MA, Meradji M, Gil-Cardoso K, Ortega N, Chomiciute G, Kharroub K. In vitro assessment of biofunctional properties of Lactiplantibacillus plantarum strain Jb21-11 and the characterization of its exopolysaccharide. Int Microbiol, 2024, 27(1239-256

[13]

Bouzaiene T, Mohamedhen Vall M, Ziadi M, Ben Rejeb I, Yangui I, Aydi A, Moktar H. Exopolysaccharides from Lactiplantibacillus plantarum C7 exhibited antibacterial, antioxidant, anti-enzymatic, and prebiotic activities. Fermentation, 2024, 10(7): 339-350

[14]

Jiang B, Wei M, He Y, Wang Y, Zhang Q, Wei H, Tao X. Antitumor effect of exopolysaccharide from Lactiplantibacillus plantarum WLPL09 on melanoma mice via regulating immunity and gut microbiota. Int J Biol Macromol, 2024, 254: 127624

[15]

Edwards U, Rogall T, Blöcker H, Emde M, Böttger EC. Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res, 1989, 17(19): 7843-7853

[16]

Li W, Mutuvulla M, Chen X, Jiang M, Dong M. Isolation and identification of high viscosity-producing lactic acid bacteria from a traditional fermented milk in Xinjiang and its role in fermentation process. Eur Food Res Technol, 2012, 235: 497-505

[17]

Xu R, Ma S, Wang Y, Liu L, Li P. Screening, identification and statistic optimization of a novel exopolysaccharide producing Lactobacillus paracasei HCT. Afr J Microbiol Res, 2010, 4(9783-795

[18]

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 1976, 72(1-2): 248-254

[19]

Ozturk S, Aslim B, Suludere Z, Tan S. Metal removal of cyanobacterial exopolysaccharides by uronic acid content and monosaccharide composition. Carbohydr Polym, 2014, 101: 265-271

[20]

Dertli E, Colquhoun IJ, Côté GL, Le Gall G, Narbad A. Structural analysis of the α-D-glucan produced by the sourdough isolate Lactobacillus brevis E25. Food Chem, 2018, 242: 45-52

[21]

Xu Y, Cui Y, Wang X, Yue F, Shan Y, Liu B, X. Purification, characterization and bioactivity of exopolysaccharides produced by Lactobacillus plantarum KX041. Int J Biol Macromol, 2019, 128: 480-492

[22]

Huang YY, Wu JM, Wu WT, Lin JW, Liang YT, Hong ZZ, Liu DM. Structural, antioxidant, and immunomodulatory activities of an acidic exopolysaccharide from Lactiplantibacillus plantarum DMDL 9010. Front Nutr, 2022, 9: 1073071

[23]

Nehal F, Sahnoun M, Smaoui S, Jaouadi B, Bejar S, Mohammed S. Characterization, high production and antimicrobial activity of exopolysaccharides from Lactococcus lactis F-mou. Microb Pathog, 2019, 132: 10-19

[24]

Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med, 1999, 26(9-10): 1231-1237

[25]

Wichienchot S, Jatupornpipat M, Rastall RA. Oligosaccharides of pitaya (dragon fruit) flesh and their prebiotic properties. Food Chem, 2010, 120(3): 850-857

[26]

Rui Y, Wan P, Chen G, Xie M, Sun Y, Zeng X, Liu Z. Simulated digestion and fermentation in vitro by human gut microbiota of intra-and extra-cellular polysaccharides from Aspergillus cristatus. Lwt, 2019, 116: 108508

[27]

Minekus M, Alminger M, Alvito P, Ballance S, Bohn TORSTEN C, Bourlieu C, Brodkorb A. A standardised static in vitro digestion method suitable for food–an international consensus. Food Funct, 2014, 5(6): 1113-1124

[28]

Ujiroghene OJ, Liu L, Zhang S, Lu J, Zhang C, Pang X, Lv J. Potent α-amylase inhibitory activity of sprouted quinoa-based yoghurt beverages fermented with selected anti-diabetic strains of lactic acid bacteria. RSC Adv, 2019, 9(179486-9493

[29]

Ogidi CO, Ubaru AM, Ladi-Lawal T, Thonda OA, Aladejana OM, Malomo O. Bioactivity assessment of exopolysaccharides produced by Pleurotus pulmonarius in submerged culture with different agro-waste residues. Heliyon, 2020, 6(12): e05685

[30]

Zhang R, Zhou Z, Ma Y, Du K, Sun M, Zhang H, Chen P. Exopolysaccharide from Lactiplantibacillus plantarum YT013 and its apoptotic activity on gastric cancer AGS cells. Fermentation, 2023, 9(6539-554

[31]

Zehir Şentürk D, Dertli E, Erten H, Şimşek Ö. Structural and technological characterization of ropy exopolysaccharides produced by Lactobacillus plantarum strains isolated from Tarhana. Food Sci Biotechnol, 2020, 29: 121-129

[32]

Yao M, Zhang M, Lai T, Yang Z. Characterization and in vitro fecal microbiota regulatory activity of a low-molecular-weight exopolysaccharide produced by Lactiplantibacillus plantarum NMGL2. Foods, 2022, 11(3): 393-411

[33]

Laws AP, Chadha MJ, Chacon-Romero M, Marshall VM, Maqsood M. Determination of the structure and molecular weights of the exopolysaccharide produced by Lactobacillus acidophilus 5e2 when grown on different carbon feeds. Carbohydr Polym, 2008, 343(2): 301-307

[34]

Fukuda K, Shi T, Nagami K, Leo F, Nakamura T, Yasuda K, Urashima T. Effects of carbohydrate source on physicochemical properties of the exopolysaccharide produced by Lactobacillus fermentum TDS030603 in a chemically defined medium. Carbohydr Polym, 2010, 79(4): 1040-1045

[35]

Ayyash M, Abu-Jdayil B, Itsaranuwat P, Galiwango E, Tamiello-Rosa C, Abdullah H, Hamed F. Characterization, bioactivities, and rheological properties of exopolysaccharide produced by novel probiotic Lactobacillus plantarum C70 isolated from camel milk. Int J Biol Macromol, 2020, 144: 938-946

[36]

Zhu Y, Wang X, Pan W, Shen X, He Y, Yin H, Liu S. Exopolysaccharides produced by yogurt-texture improving Lactobacillus plantarum RS20D and the immunoregulatory activity. Int J Biol Macromol, 2019, 121: 342-349

[37]

Yu L, Ye G, Qi X, Yang Y, Zhou B, Zhang Y, Ping W. Purification, characterization and probiotic proliferation effect of exopolysaccharides produced by Lactiplantibacillus plantarum HDC-01 isolated from sauerkraut. Front Microbiol, 2023, 14: 1210302

[38]

Zaghloul EH, Ibrahim MI. Production and characterization of exopolysaccharide from newly isolated marine probiotic Lactiplantibacillus plantarum EI6 with in vitro wound healing activity. Front Microbiol, 2022, 13: 903363

[39]

Amiri S, Mokarram RR, Khiabani MS, Bari MR, Khaledabad MA. Exopolysaccharides production by Lactobacillus acidophilus LA5 and Bifidobacterium animalis subsp. lactis BB12: optimization of fermentation variables and characterization of structure and bioactivities. Int J Biol Macromol, 2019, 123: 752-765

[40]

Mohan CC, Harini K, Aafrin BV, Babuskin S, Karthikeyan S, Sudarshan K, Sukumar M. Extraction and characterization of polysaccharides from tamarind seeds, rice mill residue, okra waste and sugarcane bagasse for its bio-thermoplastic properties. Carbohydr Polym, 2018, 186: 394-401

[41]

Zang W, Cao H, Ge J, Zhao D. Structures, physical properties and antibacterial activity of silver nanoparticles of Lactiplantibacillus plantarum exopolysaccharide. Int J Biol Macromol, 2024, 263 130083

[42]

Yang X, Ren Y, Li L. The relationship between charge intensity and bioactivities/processing characteristics of exopolysaccharides from lactic acid bacteria. LWT, 2022, 153: 112345

[43]

Ge Z, Azi F, Bao X, Yin X, Feng X, Zhang M, Dong M. Optimization and characterization of exopolysaccharides from Leuconostoc citreum BH10 and its functional properties in vitro. Food Prod Process Nutr, 2023, 5(1): 23-40

[44]

Hamidi M, Jafari H, Siminska-Stanny J, Okoro OV, Fatimi A, Shavandi A. Anionic exopolysaccharide from Cryptococcus laurentii 70766 as an alternative for alginate for biomedical hydrogels. Int J Biol Macromol, 2022, 212: 370-380

[45]

Sharma V, Ghosh M. Characterization of immunomodulatory, anticancer and antioxidant properties of an extracellular polymer produced by Enterococcus sp. in vegetable waste medium. Environ Sustain, 2021, 4(2419-428

[46]

Prajapati D, Bhatt A, Gupte A. Evaluation of bioactive attributes and emulsification potential of exopolysaccharide produced by a brown-rot fungus Fomitopsis meliae AGDP-2. Appl Biochem Biotechnol, 2023, 195(5): 2974-2992

[47]

McCallum N, Najlah M. The anticancer activity of monosaccharides: perspectives and outlooks. Cancers, 2024, 16(16): 2775-2801

[48]

Ahmad MF, Ahmad FA, Alsayegh AA, Zeyaullah M, Babalghith AO, Faidah H, Bantun F. Probiotics and cancer: mechanistic insights and organ-specific impact. Biomolecules, 2025, 15(6): 879-916

[49]

Guo R, Chen M, Ding Y, Yang P, Wang M, Zhang H, Ma H. Polysaccharides as potential anti-tumor biomacromolecules—a review. Front Nutr, 2022, 9: 838179

[50]

Bouallegue A, Casillo A, Chaari F, La Gatta A, Lanzetta R, Corsaro MM, Ellouz-Chaabouni S. Levan from a new isolated Bacillus subtilis AF17: purification, structural analysis and antioxidant activities. Int J Biol Macromol, 2020, 144: 316-324

[51]

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

[52]

Aliouche N, Sifour M, Ouled-Haddar H. Assessment of the potential bioactivities and biocompati-bility of exopolysaccharides secreted by three strains of Lactiplantibacillus plantarum isolated from infant faeces. J Biol Act Prod Nat, 2024, 14(6): 702-718

[53]

Wang C, Li W, Chen Z, Gao X, Yuan G, Pan Y, Chen H. Effects of simulated gastrointestinal digestion in vitro on the chemical properties, antioxidant activity, α-amylase and α-glucosidase inhibitory activity of polysaccharides from Inonotus obliquus. Food Res Int, 2018, 103: 280-288

[54]

Zhou X, Zhang Z, Huang F, Yang C, Huang Q. In vitro digestion and fermentation by human fecal microbiota of polysaccharides from flaxseed. Molecules, 2020, 25(19): 4354

[55]

Hongpattarakere T, Cherntong N, Wichienchot S, Kolida S, Rastall RA. In vitro prebiotic evaluation of exopolysaccharides produced by marine isolated lactic acid bacteria. Carbohydr Polym, 2012, 87(1846-852

[56]

Liang L, Liu G, Zhang F, Li Q, Linhardt RJ. Digestibility of squash polysaccharide under simulated salivary, gastric and intestinal conditions and its impact on short-chain fatty acid production in type-2 diabetic rats. Carbohydr Polym, 2020, 235: 115904

[57]

Wang T, Ye Z, Liu S, Yang Y, Dong J, Wang K, Liu D. Effects of crude Sphallerocarpus gracilis polysaccharides as potential prebiotics on acidifying activity and growth of probiotics in fermented milk. LWT, 2021, 149: 111882

[58]

Das D, Baruah R, Goyal A. A food additive with prebiotic properties of an α-d-glucan from Lactobacillus plantarum DM5. Int J Biol Macromol, 2014, 69: 20-26

[59]

Paventi G, Di Martino C, Coppola F, Iorizzo M. β-Glucosidase activity of Lactiplantibacillus plantarum: a key player in food fermentation and human health. Foods, 2025, 14(9): 1451-1479

RIGHTS & PERMISSIONS

Jiangnan University

AI Summary AI Mindmap
PDF

9

Accesses

0

Citation

Detail

Sections
Recommended

AI思维导图

/