Gastrointestinal Mikroflorayı Desteklemek İçin Bacillus clausii Sporları İçeren Probiyotik Tofu

Öz

Amaç: Bu çalışma, vegan bireylerin probiyotik tüketimi için diyetlerini değiştirmeden ve ek probiyotik hapları kullanmak zorunda kalmadan yararlanabilecekleri probiyotik Bacillus clausii sporları ile zenginleştirilmiş bir tofu ürünü geliştirmek amacıyla yapılmıştır.
Yöntem: B. clausii, 2xSG ortamında sporlanacak şekilde kültürlendi. Elde edilen B. clausii sporlarının tofuya eklenmesi, probiyotiğin tofu içinde en yüksek miktarda tutulabilmesini sağlamak için prosedürdeki optimum adımı belirlemek için iki farklı yöntemle gerçekleştirildi; i) pıhtılaştırıcı MgCl2’den önce probiyotik sporların eklenmesi (P-C tofu), ii) pıhtılaştırıcıdan sonra probiyotik sporların eklenmesi (C-P tofu).
Bulgular: P-C tofu ve C-P tofudaki probiyotik sporların verimleri sırasıyla 5,45±0,40 ve 5,33±0,56 log cfu/g olarak hesaplandı ve bu hedeflenen 6 log cfu/g seviyeye oldukça yaklaşmıştır. Probiyotik konsantrasyonu açısından P-C ve C-P tofu yöntemleri arasında anlamlı bir fark gözlenmedi (P=0,36; t testi). P-C tofunun -18°C’de saklanması probiyotik yükü 1 ayda 0,45 log cfu/g azaltmıştır (P=0,003). Soğuk zincir depolamanın bir günlük 20°C’de kesintiye uğraması 0,85 log seviyede anlamlı bir probiyotik azalmasına neden oldu (P=0,01). Sote pişirme uygulaması tofudaki spor yükünü 1,22 log seviyede anlamlı derecede azaltmıştır (P=0,004). P-C ve probiyotik içermeyen tofuların duyusal değerlendirmesinde genel kabul edilebilirlik özelliklerinde anlamlı bir farklılık gözlenmedi (P>0,05).
Sonuç: Probiyotikle zenginleştirilmiş tofu, tüketicilerin sağlığını olumlu yönde etkilerken veganlar ve vejetaryenler için sağlıklı bir alternatif seçim olabilir.

Anahtar Kelimeler

• Tofu
• Bacillus clausii
• Probiyotikler
• Zenginleştirme

Proje Numarası

1919B011801877

Probiotic Tofu with Bacillus clausii Spores to Support Gastrointestinal Microflora

Öz

Objective: This study was conducted to develop a tofu product enriched with probiotic Bacillus clausii spores that vegan individuals can benefit from without changing their diets for probiotic consumption and without having to use the supplementary probiotic pills.
Method: B. clausii was cultured to sporulate in 2xSG medium. The resulting B. clausii spores were added to tofu by two different methods to determine the optimum step in the procedure to ensure that the probiotic was able to retain in tofu at the highest amount; i) addition of probiotic spores before coagulant MgCl2 (P-C tofu), ii) addition of probiotic spores after the coagulant (C-P tofu).
Results: The yields of probiotic spores in P-C and C-P tofu were calculated to be 5.45±0.40 and 5.33±0.56 log cfu/g, respectively, which was slightly lower than the targeted level, 6 log cfu/g. No significant difference was observed between the P-C and C-P tofu methods in terms of probiotic concentration (P=0.36; t test). Storage of P-C tofu at -18°C decreased the probiotic load by 0.45 log cfu/g in 1 month (P=0.003). One-day temperature abuse at 20°C of cold chain storage resulted in a significant 0.85 log reduction (P=0.01). Sauté cooking was found to decrease the spore load in tofu significantly by 1.22 log (P=0.004). Sensory evaluation of P-C and probiotic free tofu did not reveal a significant difference in general acceptability features (P>0.05).
Conclusion: Probiotic enriched tofu may be a healthy alternative choice for vegans and vegetarians while positively affecting the health of consumers.

Anahtar Kelimeler

• Tofu
• Bacillus clausii
• Probiotics
• Fortification

Destekleyen Kurum

Tübitak BİDEB 2209-A

Proje Numarası

1919B011801877

Teşekkür

The authors would like to thank Aydın Adnan Menderes University, Science Technology Research and Application Center, where this study was performed. Authors also thank İnci Özcebeci from Vegan İşler Ltd. Şti. in Istanbul, Turkey for generously donating soy beans, Dr. Daniel R. Zeigler, Dr. Munkhtsetseg Banzragch, and Öyküsu Atılgan for their suggestions and comments.

Kaynakça

1. 1. Cutting, S. M. (2011). Bacillus probiotics. Food Microbiology. 28, 214–220
2. 2. Akan, E., and Kınık, Ö. (2015). Gıda üretimi ve depolanması sırasında probiyotiklerin canlılıklarını etkileyen faktörler - factors effecting probiotic viability during processing and storage of food. Celal Bayar Üniversitesi Fen Bilimleri Dergisi. 11, 155–166
3. 3. Thomas, D. W., Greer, F. R., American Academy of Pediatrics Committee on Nutrition, & American Academy of Pediatrics Section on Gastroenterology, Hepatology, and Nutrition (2010). Probiotics and prebiotics in pediatrics. Pediatrics, 126(6), 1217–1231.
4. 4. Rivera-Espinoza, Y., and Gallardo-Navarro, Y. (2010). Non-dairy probiotic products. Food Microbiology. 27, 1–11
5. 5. Pimentel, T. C., da Costa, W. K. A., Barão, C. E., Rosset, M., and Magnani, M. (2021). Vegan probiotic products: A modern tendency or the newest challenge in functional foods. Food Research International. 140, 110033
6. 6. Panghal, A., Janghu, S., Virkar, K., Gat, Y., Kumar, V., and Chhikara, N. (2018) Potential non-dairy probiotic products – A healthy approach. Food Bioscience. 21, 80–89
7. 7. Crittenden, R., Laitila, A., Forssell, P., Mättö, J., Saarela, M., Mattila-Sandholm, T., et al. (2001). Adhesion of bifidobacteria to granular starch and its implications in probiotic technologies. Applied and Environmental Microbiology, 67, 3469–3475
8. 8. Patrone, V., Molinari, P., and Morelli, L. (2016) Microbiological and molecular characterization of commercially available probiotics containing Bacillus clausii from India and Pakistan. International Journal of Food Microbiology. 237, 92–97

9. 9. Ploll, U., Petritz, H., and Stern, T. (2020) A social innovation perspective on dietary transitions: Diffusion of vegetarianism and veganism in Austria. Environmental Innovation and Societal Transitions. 36, 164–176
10. 10. Zimmer, J., Lange, B., Frick, J. S., Sauer, H., Zimmermann, K., Schwiertz, A., et al. (2012). A vegan or vegetarian diet substantially alters the human colonic faecal microbiota. European Journal of Clinical Nutrition. 66, 53–60
11. 11. Rajkumar, H., Mahmood, N., Kumar, M., Varikuti, S. R., Challa, H. R., and Myakala, S. P. (2014) Effect of Probiotic (VSL#3) and Omega-3 on Lipid Profile, Insulin Sensitivity, Inflammatory Markers, and Gut Colonization in Overweight Adults: A Randomized, Controlled Trial. Mediators of Inflammation. 2014, 1–8
12. 12. Kovats, S. K., Doyle, M. P., and Tanaka, N. (1984) Evaluation of the microbiological safety of tofu. Journal of Food Protection. 47, 618–622
13. 13. Jubayer, M., Uddin, M., and Faruque, M. (2014) Standardization parameters for production of tofu using WSD-Y-1 machine. Journal of the Bangladesh Agricultural University. 11, 307–312
14. 14. Duc, L. H., Hong, H. A., Barbosa, T. M., Henriques, A. O., and Cutting, S. M. (2004) Characterization of Bacillus probiotics available for human use. Applied and Environmental Microbiology. 70, 2161–2171
15. 15. Horosheva, T. V., Vodyanoy, V., and Sorokulova, I. (2014) Efficacy of Bacillus probiotics in prevention of antibiotic‐associated diarrhoea: a randomized, double‐blind, placebo‐controlled clinical trial. JMM Case Reports. 1, 3–8
16. 16. Sudha, M. R., Bhonagiri, S., and Kumar, M. A. (2013) Efficacy of Bacillus clausii strain UBBC-07 in the treatment of patients suffering from acute diarrhoea. Beneficial Microbes. 4, 211–216
17. 17. Turnbull, P. C. B. (1996) Bacillus. in S. Baron (Ed.), Medical Microbiology, 4th Ed., University of Texas Medical Branch at Galveston, Galveston, TX
18. 18. Schaeffer, P., Millet, J., and Aubert, J. P. (1965) Catabolic repression of bacterial sporulation. Proceedings of the National Academy of Sciences. 54, 704–711
19. 19. Leighton, T. J., and Doi, R. H. (1971) The stability of messenger ribonucleic acid during sporulation in Bacillus subtilis. Journal of Biological Chemistry. 246, 3189–3195
20. 20. Smith, L. S., Wallace, L., and Rastogi, V. K. (2011) Studies on Sporulation Optimization and Chracterization of Bacillus subtilis Spore Quality, Edgewood Chemical Biological Center, Maryland. https://apps.dtic.mil/sti/citations/ADA553789
21. 21. Zielinska, D., Kaminska, A., and Kolozyn-Krajewska, D. (2015) Development of tofu production method with probiotic bacteria addition. Journal of Microbiology, Biotechnology and Food Sciences. 04, 485–490
22. 22. Nielsen, P., Fritze, D., and Priest, F. G. (1995) Phenetic diversity of alkaliphilic Bacillus strains: proposal for nine new species. Microbiology. 141, 1745–1761
23. 23. Leighton, T. (1974) Further studies on the stability of sporulation messenger ribonucleic acid in Bacillus subtilis. Journal of Biological Chemistry. 249, 7808–7812
24. 24. Yano, K., Masuda, K., Akanuma, G., Wada, T., Matsumoto, T., Shiwa, Y., et al. (2016). Growth and sporulation defects in Bacillus subtilis mutants with a single rrn operon can be suppressed by amplification of the rrn operon. Microbiology. 162, 35–45
25. 25. Dey, A., Prasad, R., Kaur, S., Singh, J., and Luwang, M. D. (2017) Tofu: technological and nutritional potential. Indian Food Industry Magazine. 36, 8–24
26. 26. Chavan, S. M., and Verma, R. C. (2017) Textural and Sensory Characteristics of Freeze Dried Tofu. International Journal of Current Microbiology and Applied Sciences. 6, 119–128
27. 27. Sánchez, B., Arias, S., Chaignepain, S., Denayrolles, M., Schmitter, J. M., Bressollier, P., et al. (2009). Identification of surface proteins involved in the adhesion of a probiotic Bacillus cereus strain to mucin and fibronectin. Microbiology. 155, 1708–1716
28. 28. Chang, K. L., Lin, Y. S., and Chen, R. (2003). The effect of chitosan on the gel properties of tofu (soybean curd). Journal of Food Engineering. 57, 315–319
29. 29. Li, M., Chen, F., Yang, B., Lai, S., Yang, H., Liu, K., Bu, G., et al. (2015). Preparation of organic tofu using organic compatible magnesium chloride incorporated with polysaccharide coagulants. Food Chemistry. 167, 168–174
30. 30. Shen, Y. R., and Kuo, M. I. (2017) Effects of different carrageenan types on the rheological and water-holding properties of tofu. LWT. 78, 122–128
31. 31. Li, L., Wang, C., Li, K., Qin, W., Wu, D., Hu, B., et al. (2021). Influence of soybean protein isolate-dextran conjugates on the characteristics of glucono-δ-lactone-induced tofu. LWT. 139, 110588
32. 32. Spotti, M. J., Martinez, M. J., Pilosof, A. M. R., Candioti, M., Rubiolo, A. C., and Carrara, C. R. (2014) Influence of Maillard conjugation on structural characteristics and rheological properties of whey protein/dextran systems. Food Hydrocolloids. 39, 223–230
33. 33. Lakshmanan, R., de Lamballerie, M., and Jung, S. (2006) Effect of soybean-to-water ratio and pH on pressurized soymilk properties. Journal of Food Science. 71, E384–E391
34. 34. Lambrecht, H. S., Nielsen, S. S., Liska, B. J., and Nielsen, N. C. (1996) Effect of soybean storage on tofu and soymilk production. Journal of Food Quality. 19, 189–202
35. 35. Ng, K. H., Lye, H. S., Easa, A. M., and Liong, M. T. (2008). Growth characteristics and bioactivity of probiotics in tofu-based medium during storage. Annals of Microbiology. 58, 477–487
36. 36. Liu, D. M., Li, L., Yang, X. Q., Liang, S. Z., and Wang, J. S. (2006). Survivability of Lactobacillus rhamnosus during the preparation of soy cheese. Food Technology and Biotechnology. 44, 417– 422
37. 37. Dalev, D., Bielecka, M., Troszynska, A., Ziajka, S., and Lamparski, G. (2006) Sensory quality of new probiotic beverages based on cheese whey and soy preparation. Polish Journal of Food and Nutrition Sciences. 15, 71–77
38. 38. Kandiah, J., and Laird, J. (2002) Sensory attributes and acceptance of flavored soy nuts by college students. Nutrition Research. 22, 405–410
39. 39. Miguel, D., Valdez, G., and Rossi, E. (2004) Sensory and chemical aspects of frozen soy yogurt fermented with Enterococcus faecium and Lactobacillus jugurti. Alimentos e Nutrição Araraquara. 15, 197–201
40. 40. Wenrich, T., and Cason, K. (2004) Consumption and perceptions of soy among low-income adults. Journal of Nutrition Education and Behavior. 36, 140–145