Document Type : Research Article

Authors

1 Department of Food Science and Technology, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran.

2 Department of Food Technology and Processing Faculty, Department of Chemical Technologies, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran.

3 Biosystems Engineering Faculty, Department of Agricultural Research, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran

Abstract

Intoduction: Probiotics are defined as essential live microorganisms that, when administered in adequate amounts, confer a health benefit on the host. The range of beneficial properties reaches from lowering cholesterol to preventing cancer. The most important probiotic microorganisms belong to the group of lactic acid bacteria. Lactobacillus reuteri which naturally occurs in the human intestine possess probiotic properties with good colonization potential. The development of probiotic foods presents many challenges, particularly with respect to the stability of the bioactive compounds during processing, storage and passage through acidic gastric environment. Therefore, it is a great challenge to bring the probiotics into a stable form, which guarantees, that the microorganisms reach their target location, the human intestine, in an adequate amount. Microencapsulation helps improve survival probiotic bacteria from environmental stresses. In most studies, probiotic bacteria are entrapped in a gel matrix of natural biological materials such as alginate, or gellan. The core and wall solutions are turned into drops of the desired size by employing an emulsion method. The main problem in the probiotic entrapment approach is that gel bead entrapment technologies generally stabilize the bacteria in liquid products and are difficult to scale up. In order to extend the shelf life of encapsulated probiotics, a glassy state form of the embedding matrix is required. This can be achieved by employing such as air-suspension fluidized-bed coating. In the present research, an air-suspension fluidized-bed technique for generation of core and shell microcapsules containing probiotic Lactobacillus reuteri cells and the efficacy of shellac and sodium alginate at different concentrations on viability of capsules in simulated gastrointestinal conditions was evaluated.

Materials and methods: Pure freeze-dried Lactobacillus reuteri PT-1655 were obtained from Persian Type Culture Collection (Tehran, Iran) and were activated by inoculation in the MRS broth at 37°C for 36-48 h. The air-suspension process was performed in a Wurster coater system with a bottomspraying atomizer. The growth curve of lactobacillus reuteri were determined by measuring the optical density (turbidity) at 600 nm to estimate the time when the growth curve enters a stationary phase in which bacteria develop a general stress resistance and are thus more resistant to various types of stresses. In various pretests the fluidization pressure, the atomization pressure and the spraying rate of the microencapsulation process were varied to examine their influence on process conditions, especially on the particle development. Several different solutions of Lactobacillus reuteri were prepared and evaluated for percentage survival during the coating. The solution containing Lactobacillus reuteri (6–12 g/100 g solution), maltodextrin (4–7 g/100 g solution) and sorbitol (4–7 g/100 g solution) concentrations was spray-coated at three inlet temperatures: 37, 47 and 62°C onto and absorbed by the inert carrier microcrystalline cellulose to produce nonagglomerating dry coated. For the coating processes an aqueous shellac solution at 3 concentrations (16, 17 and 18% (w/v)), containing plasticizers in the ratios of 95 + 5 and an aqueous sodium alginate solution at 3 concentrations (0.5, 1 and 1.5% (w/v)), were used. Simulated gastric juice was prepared fresh daily containing 3.2 mg of pepsin, 1 ml of NaCl solution (0.5%) and acidified with HCl (1.2 M) to pH 1.5 ± 0.5. Tolerance to gastric juice was examined by placing freshly prepared cells in a tube containing sterile simulated gastric juice for 1 h and incubated at 37°C for 2 h. To characterize the morphology of the MCC particles coated with the different matrix formulations, SEM images were taken. Experimental data have been represented as the mean with standard deviation (SD) of different independent determinations. The significance of differences was evaluated by analysis of variance (ANOVA). Differences were considered statistically significant at p

Keywords

Bian, L., 2008, An in vitro antimicrobial and safety study of Lactobacillus reuteri DPC16 for validation of probiotic concept, Master’s thesis, Massey University, Auckland, New Zealand.
Carlise, B., Fritzen-Freire, A., Elane, S., Prudêncio, A., Renata, D., Amboni, A., Stephanie, S.& Pinto, A.,2012, Microencapsulation of bifidobacteria by spray drying in the presence of prebiotics. Food Research International, 45, 306–312.
Graff, S., Hussain, S., Chaumeil, J.C.& Charrueaul, C., 2007, Increased intestinal delivery of viable Saccharomyces boulardii by encapsulation in microspheres. Pharmaceutical Research, 25, 1290-1296.
Hamad, S.A., Stoyanov, S.D.& Paunov, V.N., 2012, Triggered cell release from shellac-cell composite microcapsules, Soft Matter, 5069-5077.
Kahm, M., AhrCampus, R.& Hasenbrink, G., 2010, grofit: Fitting Biological Growth Curves with R. Journal of Statistical Software, 33(7), 23-45.
Kartheek, A., 2011, Microencapsulation of probiotics (Lactobacillus acidophilus and Lactobacillus rhamnosus) in raspberry powder by spray drying: optimization and storage stability studies. Master’s thesis, McGill University, Quebec, Canada.
Lee, K. Y.& Heo, T., 2000, Survival of Bifidobacterium longum immobilized in calcium alginate beads in simulated gastric juices and bile salt solution. Applied and Environmental Microbiology, 66, 869–873.
Mandal, S., Puniya, K.& Singh, K., 2006, Effect of alginate concentrations on survival of microencapsulated Lactobacillus casei NCDC-298. International Dairy Journal, 126(2-3), 249-28-4.
Marinescu, D., 2012, Bile Salt Hydrolyzing Lactobacillus reuteri (NCIMB 30242) for the reduction of markers of metabolic disease. Master’s thesis, McGill University, Quebec, Canada. Meng, X.C., Stanton, C., Fitzgerald, G.F., Daly, C.& Ross, R.P., 2006, Anhydrobiotics: The challenges of drying probiotic cultures. Food Chemistry, 106, 1406–1416.
Michael, T., Cook, G., Tzortzis, D.& Charalampopoulos, V.,2010, Microencapsulation of probiotics for gastrointestinal delivery. Journal of Controlled Release, 162, 56–67.
Muthukumarasamy, P., Allan-Wojtas, P.& Holley, R.A., 2006, Stability of Lactobacillus reuteri in different types of microcapsules. Journal of Food Science, 71, 20-24.
Piar, H.& Kok, k., 2014, Enteric coating of granules containing the probiotic Lactobacillus acidophilus. Acta Pharmaceutica, 64, 247–256.
Reid, G., 2015, The growth potential for dairy probiotics. International Dairy Journal, 49: 16–22. Stephen, R.L., Werner Jim, R., Anthony, H.J., Richard, H., Archer, D.& Pearce, L., 2007, Air-suspension particle coating in the food industry: Part I — state of the art. Powder Technology, 171, 25–33.
Sabikhi, L., Babu, R., Thompkinson, D. K.& Kapila, S., 2010, Resistance of Microencapsulated Lactobacillus acidophil LA1 to Processing Treatments and Simulated Gut Conditions. Food Bioprocess Technology, 3, 586–593.
Santivarangkna, C., Kulozik, U.& Foerst, P., 2007, Alternative Drying Processes for the Industrial Preservation of Lactic Acid Starter Cultures. Biotechnology, 23, 302-315.
Sanjay, K., Jain, A., Yashwant, G.& Manisha, A., 2007, Design and Development of Hydrogel Beads for Targeted Drug Delivery to the Colon. AAPS PharmSciTech, 8 (3), 56.
Schell, D.& Beermann, C., 2014, Fluidized bed microencapsulation of Lactobacillus reuteri with sweet whey and shellac for improved acid resistance and in-vitro gastro-intestinal survival. Food Research International, 62, 308.
Semyonov, D., Ramon, O., Kovacs, A., Friedlander, L.& Shimoni, E., 2014, Air-Suspension Fluidized-Bed Microencapsulation of Probiotics. Drying Technology: An International Journal, 30(16), 1918-1930.
Semyonov, D., Ramon, O., Kaplun, Z., Levin-Brener, L., Gurevich, N.& Shimoni, E., 2010, Microencapsulation of Lactobacillus paracasei by spray freeze drying. Food Research International, 43(1), 193–20.
Sinha, A., Dudani, T.&Ranganathan, B.,2001, National dairy Research protective effect of fortified skim milk as suspending medium for freeze drying of different Lactic acid bacteria. Journal of food science, 39.
Stephan, A., 2008, Innovative product formulations applying the fluidised bed technology. Doctoral thesis, book edited by Everlon Cid Rigobelo, ISBN 978-953-51-0776-7
Stummer, S., Salar-Behzadi, S., Unger, F., M, Oelzant., Penning, M.& Viernstein, H.,2010, Application of shellac for the development of probiotic formulations. Food Research International, 43, 1312–1320.
Sultana, K., Godward, G., Reynolds, N., Arumugaswamy, R., Peiris, P.& Kailasapathy, K., 2000, Encapsulatio of probiotic bacteria with alginate-starch and evaluation of survival in simulated gastrointestinal conditions and in yoghurt. International Journal of Food Microbiology, 62(1-2), 47-55.
Wagner, R., Shemedia, J.& Johnson, L., 2012, Protection of Vaginal Epithelial Cells with Probiotic Lactobacilli and the Effect of Estrogen against Infection by Candida albicans Microbiology Division, National Center for Toxicological Research, Jefferson, USA. Open Journal of Medical Microbiology, 2, 54-64
Wolfgang, K., Andreas, R.& Klaus, D., 2000, In vitro growth behaviour of probiotic bacteria in culture media with carbohydrates of prebiotic importance. Microbial Ecology in Health and Disease,12, 27–34.
Wurster, E., 1959, Air-Suspension Technique of Coating Drug Particles. Journal of the American Pharmaceuticasaslociation, XLVIII, 8
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