Abstract
The establishment of the health-promoting benefits of probiotics is challenged by the antimicrobial bio-barriers throughout the host’s gastrointestinal (GI) tract after oral administration. Although microencapsulation has been frequently utilised to enhance the delivery of probiotics, microcapsules of sub-100 μm were found to be ineffective and therefore questioned as an effective delivery vehicle for viable probiotics despite the sensory advantage. In this study, four probiotics strains were encapsulated in chitosan-coated alginate microcapsules of sub-100 μm. Only a minor protective effect was observed from this original type of microcapsule. In order to enhance the survival of these probiotics, sucrose, a metabolisable sugar, and lecithin vesicles were added to the wall material. Both of the ingredients could be readily encapsulated with the probiotics, and protected them from stresses in the simulated GI fluids. The metabolisable sugar effectively increased the survival of the probiotics in gastric acid, mainly through energizing the membrane-bound F1F0-ATPases. The lecithin vesicles proved to alleviate the bile salt stress, and hence notably reduced the viability loss at the elevated bile salt concentrations. Overall, three out of the total four probiotics in the reinforced sub-100 μm microencapsules could significantly survive through an 8-h sequential treatment of the simulated GI fluids, giving less than 1-log drop in viable count. The most vulnerable strain of bifidobacteria also yielded a viability increase of 3-logs from this protection. In conclusion, the sub-100 μm microcapsules can be a useful vehicle for the delivery of probiotics, as long as suitable protectants are incorporated in the wall matrix.
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References
Anal A, Singh H (2007) Recent advances in microencapsulation of probiotics for industrial applications and targeted delivery. Trends in Food Science & Technology 18(5):240–251
Anal AK, Bhopatkar D, Tokura S, Tamura H, Stevens WF (2003) Chitosan-alginate multilayer beads for gastric passage and controlled intestinal release of protein. Drug Development and Industrial Pharmacy 29(6):713–724. doi:10.1081/DDC-120021320
Begley M, Gahan C, Hill C (2005) The interaction between bacteria and bile. FEMS Microbiology Reviews 29(4):625–651
Chandramouli V, Kailasapathy K, Peiris P, Jones M (2004) An improved method of microencapsulation and its evaluation to protect Lactobacillus spp. in simulated gastric conditions. Journal of Microbiological Methods 56(1):27–35
Chen X, Sun Z, Meng H, Zhang H (2009) The acid tolerance association with expression of H+-ATPase in Lactobacillus casei. International Journal of Dairy Technology 62(2):272–276
Corcoran B, Stanton C, Fitzgerald G, Ross R (2005) Survival of probiotic lactobacilli in acidic environments is enhanced in the presence of metabolizable sugars. Applied and Environmental Microbiology 71(6):3060
Cotter P, Hill C (2003) Surviving the acid test: responses of gram-positive bacteria to low pH. Microbiology and Molecular Biology Reviews 67(3):429
Cotter P, Gahan C, Hill C (2000) Analysis of the role of the Listeria monocytogenes F0F1-ATPase operon in the acid tolerance response. International Journal of Food Microbiology 60(2–3):137–146
Cui J, Goh J, Kim P, Choi S, Lee B (2000) Survival and stability of bifidobacteria loaded in alginate poly-l-lysine microparticles. International Journal of Pharmaceutics 210(1–2):51–59
de Vrese M, Marteau PR (2007) Probiotics and prebiotics: effects on diarrhea. Journal of Nutrition 137(3):803S–811
Donovan J, Jackson A (1993) Rapid determination by centrifugal ultrafiltration of inter-mixed micellar/vesicular (non-lecithin-associated) bile salt concentrations in model bile: influence of Donnan equilibrium effects. The Journal of Lipid Research 34(7):1121
Drouault S, Corthier G, Ehrlich SD, Renault P (1999) Survival, physiology, and lysis of Lactococcus lactis in the digestive tract. Applied and Environmental Microbiology 65(11):4881–4886
Gänzle M, Hertel C, van der Vossen J, Hammes W (1999) Effect of bacteriocin-producing lactobacilli on the survival of Escherichia coli and Listeria in a dynamic model of the stomach and the small intestine. International Journal of Food Microbiology 48(1):21–35
Giannella RA, Broitman SA, Zamcheck N (1972) Gastric acid barrier to ingested microorganisms in man: studies in vivo and in vitro. Gut 13(4):251–256. doi:10.1136/gut.13.4.251
Hansen L, Allan-Wojtas P, Jin Y, Paulson A (2002) Survival of Ca-alginate microencapsulated Bifidobacterium spp. in milk and simulated gastrointestinal conditions. Food Microbiology 19(1):35–45
Hatakka K, Mutanen M, Holma R, Saxelin M, Korpela R (2008) Lactobacillus rhamnosus LC705 together with Propionibacterium freudenreichii ssp shermanii JS administered in capsules is ineffective in lowering serum lipids. Journal of the American College of Nutrition 27(4):441
Hope M, Bally M, Webb G, Cullis P (1985) Production of large unilamellar vesicles by a rapid extrusion procedure. Characterization of size distribution, trapped volume and ability to maintain a membrane potential. Biochimica et Biophysica Acta (BBA)-Biomembranes 812 (1):55–65
Kekkonen R, Lummela N, Karjalainen H, Latvala S, Tynkkynen S, Jarvenpaa S, Kautiainen H, Julkunen I, Vapaatalo H, Korpela R (2008) Probiotic intervention has strain-specific anti-inflammatory effects in healthy adults. World Journal of Gastroenterology 14(13). doi:10.3748/wjg.14.2029
Larisch B, Poncelet D, Champagne C, Neufeld R (1994) Microencapsulation of Lactococcus lactis subsp. cremoris. Journal of Microencapsulation 11(2):189–195
Lee K, 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(2):869
Leroy F, Falony G, Vuyst L (2008) Latest developments in probiotics. In: Toldrá F (ed) Meat biotechnology. Springer, New York, pp 217–229
Li X-Y, Jin L-J, McAllister TA, Stanford K, Xu J-Y, Lu Y-N, Zhen Y-H, Sun Y-X, Xu Y-P (2007) Chitosan-alginate microcapsules for oral delivery of egg yolk immunoglobulin (IgY). Journal of Agricultural and Food Chemistry 55(8):2911–2917. doi:10.1021/jf062900q
Lorca G, Font de Valdez G (2001) Acid tolerance mediated by membrane ATPases in Lactobacillus acidophilus. Biotechnology Letters 23(10):777–780
Martin CC, Donald MS (1970) The characteristics of mixed micellar solutions with particular reference to bile. The American Journal of Medicine 49(5):590–608
Masco L, Crockaert C, Van Hoorde K, Swings J, Huys G (2007) In vitro assessment of the gastrointestinal transit tolerance of taxonomic reference strains from human origin and probiotic product isolates of Bifidobacterium. Journal of Dairy Science 90(8):3572–3578. doi:10.3168/jds.2006-548
Narain P, DeMaria E, Heuman D (1998) Lecithin protects against plasma membrane disruption by bile salts. Journal of Surgical Research 78(2):131–136
Nichols JW, Ozarowski J (1990) Sizing of lecithin-bile salt mixed micelles by size-exclusion high-performance liquid chromatography. Biochemistry 29(19):4600–4606. doi:10.1021/bi00471a014
O’Flaherty S, Goh YJ, Klaenhammer TR (2009) Genomics of probiotic bacteria. In: Charalampopoulos D, Rastall RA (eds) Prebiotics and probiotics science and technology. Springer, New York, pp 681–723
Oelschlaeger T (2010) Mechanisms of probiotic actions—a review. International Journal of Medical Microbiology 300(1):57–62
Ooi L, Liong M (2010) Cholesterol-lowering effects of probiotics and prebiotics: a review of in vivo and in vitro findings. International Journal of Molecular Sciences 11(6):23. doi:10.3390/ijms11062499
Ouwehand AC, Tölkkö S, Salminen S (2001) The effect of digestive enzymes on the adhesion of probiotic bacteria in vitro. Journal of Food Science 66(6):856–859. doi:10.1111/j.1365-2621.2001.tb15186.x
Sagawa H, Tazuma S, Kajiyama G (1993) Protection against hydrophobic bile salt-induced cell membrane damage by liposomes and hydrophilic bile salts. American Journal of Physiology- Gastrointestinal and Liver Physiology 264(5):835
Sánchez B, De Los Reyes-Gavilán CG, Margolles A (2006) The F1F0-ATPase of Bifidobacterium animalis is involved in bile tolerance. Environmental Microbiology 8(10):1825–1833. doi:10.1111/j.1462-2920.2006.01067.x
Shimakawa Y, Matsubara S, Yuki N, Ikeda M, Ishikawa F (2003) Evaluation of Bifidobacterium breve strain Yakult-fermented soymilk as a probiotic food. International Journal of Food Microbiology 81(2):131–136
Silva C, Ribeiro A, Figueiredo M, Ferreira D, Veiga F (2005) Microencapsulation of hemoglobin in chitosan-coated alginate microspheres prepared by emulsification/internal gelation. The AAPS Journal 7(4):903–913
Vanderpool C, Yan F, Polk DB (2008) Mechanisms of probiotic action: implications for therapeutic applications in inflammatory bowel diseases. Inflammatory Bowel Diseases 14(11):1585–1596. doi:10.1002/ibd.20525
Waddington L, Cyr T, Hefford M, Hansen L, Kalmokoff M (2010) Understanding the acid tolerance response of bifidobacteria. Journal of Applied Microbiology 108(4):1408–1420
Acknowledgements
This work was supported by Technology for Industry Fellowship (TIF) New Zealand grant DPAC0702 and by the Drapac Trust, New Zealand. We also thank Dr. Sabrina Tian, the team of Seafood Unit—Plant and Food Research, the team of AnQual Lab—the University of Auckland, Ms. Vivien Zhang and Ms. Yu Cao for their kind help and emotional support throughout the project.
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Chen, S., Zhao, Q., Ferguson, L.R. et al. Development of a novel probiotic delivery system based on microencapsulation with protectants. Appl Microbiol Biotechnol 93, 1447–1457 (2012). https://doi.org/10.1007/s00253-011-3609-4
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DOI: https://doi.org/10.1007/s00253-011-3609-4