Spor ve vejetatif formlarda Bacillus coagulans GBI-30 6086 ve Saccharomyces boulardii içeren fesleğen tohumu gamı bazlı probiyotik filmlerin fizikokimyasal özellikleri, depolama stabilitesi ve antibakteriyel aktivitesi

Abstract

Bu çalışma, gıda ambalajı için sürdürülebilir bir alternatif olarak fesleğen tohumu gamı (FTG) bazlı probiyotik filmlerin geliştirilmesini araştırmış ve bu filmlerin fizikokimyasal, yapısal ve biyolojik özelliklerini değerlendirmiştir. FTG ekstraksiyonunun ardından, gliserol ile plastikleştirilmiş biyopolimer filmler hazırlanmış ve Bacillus coagulans GBI-30 6086 (BC; spor ve vejetatif formlar) ile Saccharomyces boulardii ilave edilmiştir. Daha sonra filmler; nem içeriği, kalınlık, suda çözünürlük, su buharı geçirgenliği (SBG), renk parametreleri, antioksidan kapasite ve yüzey morfolojisi açısından karakterize edilmiştir. Elde edilen sonuçlar, probiyotik ilavesinin filmlerin bariyer özellikleri ve mikroyapısı üzerinde etkili olduğunu ve bu etkinin mikrobiyal forma bağlı olarak değiştiğini göstermiştir. BC’nin hem spor hem de vejetatif formları, S. boulardii ve kontrol örneğine kıyasla daha yüksek SBG değerlerine yol açarken, vejetatif form aynı zamanda en yüksek film kalınlığına neden olmuştur. Buna karşılık, S. boulardii daha düşük kalınlık ve azalmış geçirgenlik ile ilişkilendirilmiş olup, daha kompakt bir matris oluşumuna işaret etmiştir. Yirmi bir günlük depolama süresi boyunca BC’nin spor formu en yüksek canlılığı (9.14 log KOB/g) korumuş, bu durum dormansi ve direnç ile ilişkili üstün stabilitesini ortaya koyarken, vejetatif hücreler ve S. boulardii zamanla belirgin şekilde daha düşük canlılık göstermiştir. Probiyotik içeren tüm filmler Klebsiella pneumoniae ve Staphylococcus carnosus’a karşı antibakteriyel aktivite sergilemiş; ancak kontrol formülasyonunun daha yüksek inhibisyon zonları göstermesi, antibakteriyel etkinin esas olarak FTG matrisinin intrinsik özelliklerinden kaynaklandığını göstermiştir. Atomik kuvvet mikroskobu analizleri, probiyotik ilavesinin yapısal bütünlüğü bozmaksızın nanoskalada yüzey pürüzlülüğünü artırdığını ortaya koymuştur. Genel olarak bu bulgular, probiyotiklerle zenginleştirilmiş FTG bazlı filmlerin, gıda kalitesini artırma potansiyeline sahip ve fonksiyonel biyoaktifler için taşıyıcı olarak kullanılabilen çevre dostu bir aktif ambalaj sistemi olduğunu göstermektedir.

Keywords

• Fesleğen tohumu gamı
• Probiyotik film
• Bacillus coagulans
• Saccharomyces boulardii
• Aktif ambalajlama
• Antibakteriyel aktivite

Supporting Institution

Bu araştırma herhangi bir dış finansman desteği almamıştır.

Ethical Statement

Bu çalışma insan veya hayvan denekleri içermemektedir ve bu nedenle etik kurul onayı gerekmemiştir.

Thanks

Yazarların beyan edecekleri herhangi bir teşekkür bulunmamaktadır.

Basil Seed Gum-Based Probiotic Films Containing Bacillus coagulans GBI-30, 6086 Spore and Vegetative Forms and Saccharomyces boulardii: Physicochemical Properties, Storage Stability, and Antibacterial Activity

Abstract

This study investigated the development of basil seed gum (BSG)-based probiotic films as a sustainable alternative for food packaging and evaluated their physicochemical, structural, and biological properties. Following BSG extraction, glycerol-plasticized biopolymer films were prepared and incorporated with Bacillus coagulans GBI-30, 6086 (BC; spore and vegetative forms) and Saccharomyces (S.) boulardii. The films were then characterized in terms of moisture content, thickness, water solubility, water vapor permeability (WVP), color parameters, antioxidant capacity, and surface morphology. The results showed that probiotic incorporation influenced the barrier properties and microstructure of the films, with effects varying depending on the microbial form. Both spore and vegetative forms of BC resulted in higher WVP compared to that of S. boulardii and the control, while the vegetative form also led to the highest film thickness. In contrast, S. boulardii was associated with lower thickness and reduced permeability, suggesting the formation of a more compact matrix. During 21 days of storage, the spore form of BC maintained the highest viability (9.14 log CFU/g), indicating its superior stability associated with dormancy and resistance, whereas vegetative cells and S. boulardii exhibited markedly lower viability over time. All probiotic-containing films exhibited antibacterial activity against Klebsiella pneumoniae and Staphylococcus carnosus; however, the control formulation showed higher inhibition zones, indicating that the antibacterial effect was mainly associated with the intrinsic properties of the BSG matrix. Atomic force microscopy analysis demonstrated that probiotic incorporation increased nanoscale surface roughness without compromising structural integrity. Overall, these results highlight the potential of probiotic-enriched BSG films as an environmentally friendly active packaging system capable of enhancing food quality and serving as a carrier for functional bioactives.

Keywords

• Basil seed gum
• Probiotic film
• Bacillus coagulans
• Saccharomyces boulardii
• Active packaging
• Antibacterial activity

Supporting Institution

This research received no external funding.

Ethical Statement

This study did not involve human or animal subjects, and therefore no ethical approval was required.

Thanks

The authors have no acknowledgments to declare.

References

1. Adibpour, N., Hosseininezhad, M., Pahlevanlo, A., & Hussain, M. A. (2019). A review on Bacillus coagulans as a spore-forming probiotic. Applied Food Biotechnology. 61(2), 91-100. National Nutrition and Food Technology Research Institute. https://doi.org/10.22037/afb.v6i2.23958
2. Ahire, J. J., Kashikar, M. S., & Madempudi, R. S. (2021). Comparative accounts of probiotic properties of spore and vegetative cells of Bacillus clausii UBBC07 and in silico analysis of probiotic function. 3 Biotech, 11(3). https://doi.org/10.1007/s13205-021-02668-0
3. Aslan Kaya, D., Ceylan, H. G., & Atasoy, A. F. (2025). Probiotic edible films supplemented with psyllium (Plantago ovata) mucilage containing Lacticaseibacillus rhamnosus GG: coating for fresh apple slices. Food and Bioprocess Technology, 18(2), 1692-1706. https://doi.org/10.1007/s11947-024-03556-0
4. Azarashkan, Z., Farahani, S., Abedinia, A., Akbarmivehie, M., Motamedzadegan, A., Heidarbeigi, J., & Hayaloğlu, A. A. (2022). Co-encapsulation of broccoli sprout extract nanoliposomes into basil seed gum: effects on in vitro antioxidant, antibacterial and anti-Listeria activities in ricotta cheese. International Journal of Food Microbiology, 376. https://doi.org/10.1016/j.ijfoodmicro.2022.109761
5. Cinbas, A., & Yazici, F. (2008). Effect of the addition of blueberries on selected physicochemical and sensory properties of yoghurts. Food Technology and Biotechnology, 46(4), 434-441.
6. Czerucka, D., Piche, T., & Rampal, P. (2007). Review article: Yeast as probiotics-Saccharomyces boulardii. Alimentary Pharmacology and Therapeutics 26(6), 767-778. https://doi.org/10.1111/j.1365-2036.2007.03442.x
7. Davachi, S. M., Pottackal, N., Torabi, H., & Abbaspourrad, A. (2021). Development and characterization of probiotic mucilage based edible films for the preservation of fruits and vegetables. Scientific Reports, 11(1). https://doi.org/10.1038/s41598-021-95994-5
8. Ebrahimi, B., Mohammadi, R., Rouhi, M., Mortazavian, A. M., Shojaee-Aliabadi, S., & Koushki, M. R. (2018). Survival of probiotic bacteria in carboxymethyl cellulose-based edible film and assessment of quality parameters. LWT, 87, 54-60. https://doi.org/10.1016/j.lwt.2017.08.066

9. Escamilla-García, M., Calderón-Domínguez, G., Chanona-Pérez, J. J., Farrera-Rebollo, R. R., Andraca-Adame, J. A., Arzate-Vázquez, I., Mendez-Mendez, J. V., & Moreno-Ruiz, L. A. (2013). Physical and structural characterisation of zein and chitosan edible films using nanotechnology tools. International Journal of Biological Macromolecules, 61, 196-203. https://doi.org/10.1016/j.ijbiomac.2013.06.051
10. Gagliarini, N., Diosma, G., Garrote, G. L., Abraham, A. G., & Piermaria, J. (2019). Whey protein-kefiran films as driver of probiotics to the gut. LWT, 105, 321-328. https://doi.org/10.1016/j.lwt.2019.02.023
11. Ghasemlou, M., Khodaiyan, F., Oromiehie, A., & Yarmand, M. S. (2011). Development and characterisation of a new biodegradable edible film made from kefiran, an exopolysaccharide obtained from kefir grains. Food Chemistry, 127(4), 1496-1502. https://doi.org/10.1016/j.foodchem.2011.02.003
12. Gholam-Zhiyan, A., Amiri, S., Rezazadeh-Bari, M., & Pirsa, S. (2021). Stability of Bacillus coagulans IBRC-M 10807 and Lactobacillus plantarum PTCC 1058 in milk proteins concentrate (MPC)-based edible film. Journal of Packaging Technology and Research, 5(1), 11-22. https://doi.org/10.1007/s41783-021-00106-3
13. Goktas, H., Dertli, E., & Sagdic, O. (2021). Comparison of functional characteristics of distinct Saccharomyces boulardii strains isolated from commercial food supplements. LWT, 136. https://doi.org/10.1016/j.lwt.2020.110340
14. Gryciuk, A., Milner-Krawczyk, M., Rogalska, M., Banach, A. K., Sitkiewicz, E., Bakun, M., Świadek, M. E., & Mierzejewska, J. (2025). Characteristics of two Saccharomyces cerevisiae strains and their extracellular vesicles as new candidates for probiotics. probiotics and antimicrobial proteins. https://doi.org/10.1007/s12602-025-10624-0
15. Hashemi Gahruie, H., Ziaee, E., Eskandari, M. H., & Hosseini, S. M. H. (2017). Characterization of basil seed gum-based edible films incorporated with Zataria multiflora essential oil nanoemulsion. Carbohydrate Polymers, 166, 93-103. https://doi.org/10.1016/j.carbpol.2017.02.103
16. Hashemi, S. M. B., & Mousavi Khaneghah, A. (2017). Characterization of novel basil-seed gum active edible films and coatings containing oregano essential oil. Progress in Organic Coatings, 110, 35-41. https://doi.org/10.1016/j.porgcoat.2017.04.041
17. Heenan, C. N., Adams, M. C., Hosken, R. W., & Fleet, G. H. (2004). Survival and sensory acceptability of probiotic microorganisms in a nonfermented frozen vegetarian dessert. LWT, 37(4), 461-466. https://doi.org/10.1016/j.lwt.2003.11.001
18. Huang, X., Du, L., Li, Z., Yang, Z., Xue, J., Shi, J., Tingting, S., Zhai, X., Zhang, J., Capanoglu, E., Zhang, N., Sun, W., & Zou, X. (2024). Lactobacillus bulgaricus-loaded and chia mucilage-rich gum arabic/pullulan nanofiber film: An effective antibacterial film for the preservation of fresh beef. International Journal of Biological Macromolecules, 266. https://doi.org/10.1016/j.ijbiomac.2024.131000
19. Jafari, M., Mortazavian, A. M., Hosseini, H., Safaei, F., Mousavi Khaneghah, A., & Sant’Ana, A. S. (2017). Probiotic Bacillus: Fate during sausage processing and storage and influence of different culturing conditions on recovery of their spores. Food Research International, 95, 46-51. https://doi.org/10.1016/j.foodres.2017.03.001
20. Kafashan, A., & Babaei, A. (2024). Development and investigation of a polysaccharide ternary nanocomposite based on basil seed gum/graphene oxide/anthocyanin for intelligent food packaging. International Journal of Biological Macromolecules, 280. https://doi.org/10.1016/j.ijbiomac.2024.135537
21. Kanmani, P., & Lim, S. T. (2013). Development and characterization of novel probiotic-residing pullulan/starch edible films. Food Chemistry, 141(2), 1041-1049. https://doi.org/10.1016/j.foodchem.2013.03.103
22. Khazaei, N., Esmaiili, M., Djomeh, Z. E., Ghasemlou, M., & Jouki, M. (2014). Characterization of new biodegradable edible film made from basil seed (Ocimum basilicum L.) gum. Carbohydrate Polymers, 102(1), 199-206. https://doi.org/10.1016/j.carbpol.2013.10.062
23. Khodaei, D., Hamidi-Esfahani, Z., & Lacroix, M. (2020). Gelatin and low methoxyl pectin films containing probiotics: Film characterization and cell viability. Food Bioscience, 36. https://doi.org/10.1016/j.fbio.2020.100660
24. Li, L., Liu, B., Cao, J., Zhang, H., Tian, F., Yu, L., Chen, W., & Zhai, Q. (2022). Different effects of Bacillus coagulans vegetative cells and spore isolates on constipation-induced gut microbiota dysbiosis in mice. Food and Function, 13(18), 9645-9657. https://doi.org/10.1039/d2fo01668k
25. Liu, S., Cai, H., Zhang, J., Peng, X., Li, P., Guo, X., Liu, Y., Zhao, J., Yan, X., & Zhu, K. (2026). Novel basil seed gum/gelatin composite films incorporating tea polyphenols: Preparation, characterization and application in pork shelf-life extension. Materials Today Communications, 52. https://doi.org/10.1016/j.mtcomm.2026.115024
26. Mehdipour Biregani, Z., Gharachorloo, M., Daneshi, M., & Moslehi Shad, M. (2025). Evaluation of reduced-fat white cheese properties containing a concentrated emulsion stabilized with basil seed gum during shelf-life. Food Chemistry, 496. https://doi.org/10.1016/j.foodchem.2025.146773
27. Miranda, J. S., Costa, B. V., de Oliveira, I. V., de Lima, D. C. N., Martins, E. M. F., de Castro Leite Júnior, B. R., Almeida do Nascimento Benevenuto, W. C., Campelo de Queiroz, I., Ribeiro da Silva, R., & Martins, M. L. (2020). Probiotic jelly candies enriched with native Atlantic Forest fruits and Bacillus coagulans GBI-30 6086. LWT, 126. https://doi.org/10.1016/j.lwt.2020.109275
28. Mokhtaran, L., Sabahi, S., Noshad, M., & Noori, S. M. A. (2025). Investigating the effects of basil (Ocimum basilicum L.) mucilage and cellulose nanofiber-based edible films with post-probiotics on fish fillet shelf life. Applied Food Research, 5(1). https://doi.org/10.1016/j.afres.2025.100704
29. Mortazavi Moghadam, F. A., Khoshkalampour, A., Mortazavi Moghadam, F. A., PourvatanDoust, S., Naeijian, F., & Ghorbani, M. (2023). Preparation and physicochemical evaluation of casein/basil seed gum film integrated with guar gum/gelatin based nanogel containing lemon peel essential oil for active food packaging application. International Journal of Biological Macromolecules, 224, 786-796. https://doi.org/10.1016/j.ijbiomac.2022.10.166
30. Mushtaq, M., Gani, A., Gani, A., Punoo, H. A., & Masoodi, F. A. (2018). Use of pomegranate peel extract incorporated zein film with improved properties for prolonged shelf life of fresh Himalayan cheese (Kalari/kradi). Innovative Food Science and Emerging Technologies, 48, 25-32. https://doi.org/10.1016/j.ifset.2018.04.020
31. Niro, C. M., Medeiros, J. A. de, Bresolin, J. D., Dionísio, A. P., Salgaço, M. K., Sivieri, K., & Azeredo, H. M. C. (2022). Banana leathers as influenced by polysaccharide matrix and probiotic bacteria. Food Hydrocolloids for Health, 2. https://doi.org/10.1016/j.fhfh.2022.100081
32. Oliveira-Alcântara, A. V., Abreu, A. A. S., Gonçalves, C., Fuciños, P., Cerqueira, M. A., Gama, F. M. P., Pastrana, L. M., Rodrigues, S., & Azeredo, H. M. C. (2020). Bacterial cellulose/cashew gum films as probiotic carriers. LWT, 130. https://doi.org/10.1016/j.lwt.2020.109699
33. Oraç, A., Göktepe, Ç. K., Demirci, T., & Akın, N. (2025). Quality and shelf-life improvement of Kashar cheese using basil seed gum-based coatings and films. Mljekarstvo, 75(4), 217-230. https://doi.org/10.15567/mljekarstvo.2025.0402
34. Oraç, A., Konak Göktepe, Ç., Demirci, T., & Akın, N. (2023). Biodegradable Edible Film Based on Basil Seed Gum: The Effect of Gum and Plasticizer Concentrations. Journal of Polymers and the Environment, 31(11), 5003-5014. https://doi.org/10.1007/s10924-023-02923-w
35. Piermaria, J., Diosma, G., Aquino, C., Garrote, G., & Abraham, A. (2015). Edible kefiran films as vehicle for probiotic microorganisms. Innovative Food Science and Emerging Technologies, 32, 193-199. https://doi.org/10.1016/j.ifset.2015.09.009
36. Rahma, L. D., Srisa, A., Wongphan, P., Nakphaichit, M., Sablani, S. S., & Harnkarnsujarit, N. (2025). Pregelatinized starch-based edible films as effective carriers for Bacillus coagulans: influence of starch type on film properties and probiotic viability. Foods, 14(14). https://doi.org/10.3390/foods14142424
37. Romano, N., Tavera-Quiroz, M. J., Bertola, N., Mobili, P., Pinotti, A., & Gómez-Zavaglia, A. (2014). Edible methylcellulose-based films containing fructo-oligosaccharides as vehicles for lactic acid bacteria. Food Research International, 64, 560-566. https://doi.org/10.1016/j.foodres.2014.07.018
38. Sahraeian, S., Hashemi, H., Ghiasi, F., Golmakani, M. T., Valizadeh, S., Tahergorabi, R., Firsukova, Y. A., & Mousavi Khaneghah, A. (2025). Recent trends in basil seed gum-based films and coatings for food packaging: An updated review. Food Chemistry: X, 30. https://doi.org/10.1016/j.fochx.2025.102937
39. Salimiraad, S., Safaeian, S., Basti, A. A., Khanjari, A., & Nadoushan, R. M. (2022). Characterization of novel probiotic nanocomposite films based on nano chitosan/ nano cellulose/ gelatin for the preservation of fresh chicken fillets. LWT, 162. https://doi.org/10.1016/j.lwt.2022.113429
40. Semwal, A., Ambatipudi, K., & Navani, N. K. (2022). Development and characterization of sodium caseinate based probiotic edible film with chia mucilage as a protectant for the safe delivery of probiotics in functional bakery. Food Hydrocolloids for Health, 2. https://doi.org/10.1016/j.fhfh.2022.100065
41. Shaahbaz, M. S., & Jouki, M. (2024). Active microcapsules and edible films obtained from basil seed gum and ginger essential oil: fabrication and characterization. Food Biophysics, 19(4), 1192-1209. https://doi.org/10.1007/s11483-024-09888-w
42. Shahabi, N., Soleimani, S., & Ghorbani, M. (2023). Investigating functional properties of halloysite nanotubes and propolis used in reinforced composite film based on soy protein/basil seed gum for food packaging application. International Journal of Biological Macromolecules, 231. https://doi.org/10.1016/j.ijbiomac.2023.123350
43. Shahrampour, D., Khomeiri, M., Kashiri, M., & Razavi, S. M. A. (2024). Embedding probiotic bacteria into the edible films: antibacterial activity and survival in simulated gastrointestinal condition. Journal of Food Measurement and Characterization, 18(3), 1947-1957. https://doi.org/10.1007/s11694-023-02291-y
44. Shakouri, M., Salami, M., Lim, L. T., Ekrami, M., Mohammadian, M., Askari, G., Emam-Djomeh, Z., & McClements, D. J. (2023). Development of active and intelligent colorimetric biopolymer indicator: anthocyanin-loaded gelatin-basil seed gum films. Journal of Food Measurement and Characterization, 17(1), 472-484. https://doi.org/10.1007/s11694-022-01640-7
45. Soukoulis, C., Behboudi-Jobbehdar, S., Macnaughtan, W., Parmenter, C., & Fisk, I. D. (2017). Stability of Lactobacillus rhamnosus GG incorporated in edible films: Impact of anionic biopolymers and whey protein concentrate. Food Hydrocolloids, 70, 345-355. https://doi.org/10.1016/j.foodhyd.2017.04.014
46. Suwanangul, S., Jaichakan, P., Narkprasom, N., Kraithong, S., Narkprasom, K., & Sangsawad, P. (2023). Innovative ınsights for establishing a synbiotic relationship with Bacillus coagulans: viability, bioactivity, and ın vitro-simulated gastrointestinal digestion. Foods, 12(19). https://doi.org/10.3390/foods12193692
47. Tan, X., Sun, A., Cui, F., Li, Q., Wang, D., Li, X., & Li, J. (2024). The physicochemical properties of cassava starch/carboxymethyl cellulose sodium edible film incorporated of Bacillus and its application in salmon fillet packaging. Food Chemistry: X, 23. https://doi.org/10.1016/j.fochx.2024.101537
48. Tomicic, Z., Colovic, R., Cabarkapa, I., Vukmirovic, D., Djuragic, O., & Tomicic, R. (2016). Beneficial properties of probiotic yeast Saccharomyces boulardii. Food and Feed Research, 43(2), 103-110. https://doi.org/10.5937/ffr1602103t