Microencapsulation of Probiotic <em>Lactobacillus casei </em>based on Alginate and Chitosan Materials


  • Djaenudin Research Unit for Clean Technology, Indonesian Institute of Sciences, Indonesia
  • Endang Saepudin Department of Chemistry, University of Indonesia, Indonesia
  • Muhamad Nasir Research Unit for Clean Technology, Indonesian Institute of Sciences, Indonesia




Lactobacillus casei; alginate; chitosan.


Probiotics are microorganisms that live in the stomach related plot which can give health advantages to the body. Probiotics must be able to survive in an acidic environment (pH = 2) during transit in the gastric to arrive the large intestine in adequate amounts (106 - 107 CFU/g) to allow for colonization and reproducing. However, most of these probiotics cannot survive in adequate amountsin acidic conditions. One way to protect probiotic cells under these conditions is through a microencapsulation system. Microencapsulation is done to protect probiotic cells from low pH, bile salts, etc. Lactobacillus casei includes probiotics that are very sensitive to pH 2-2.5. The non-encapsulated Survivability of L. casei is only 40.14% in liquid pH 2 from 8.75log CFU/g to 3.53log CFU/g while L. casei carried out microencapsulation can achieve survivability of 63.47% to 95.3%. Alginate is a microencapsulation material that is inexpensive and sensitive to changes in pH so that it is suitable as a microencapsulation of probiotics. The surface of porous alginate needs other ingredients that can be requested by the pores. Alginate is resistant to pH 2-2.5 and expands at neutral pH to alkaline, which results in increased alginate pores. Chitosan can be used as an alginate mixture in probiotic microencapsulation materials. Chitosan can bind by crosslinking with alginate, which is between the NH2 group of chitosan and COO- group of alginate.


Author Biography

Djaenudin , Research Unit for Clean Technology, Indonesian Institute of Sciences, Indonesia

Research Unit For Clean Technology


A. Lourens-Hattingh and B. C. Viljoen, “Yogurt as probiotic carrier food,” Int. Dairy J., vol. 11, no. 1–2, pp. 1–17, 2001, doi: 10.1016/S0958-6946(01)00036-X.

L. P. Morelli and L. M. Capurso, “FAO / WHO Guidelines on Probiotics : 10 Years Later,” 2012. doi: https://doi.org/10.1097/MCG.0b013e318269fdd5.

W.K. Ding and N. P. Shah, “An Improved method of microencapsulation of probiotic bacteria for their stability in acidic,” Food Microbiol. Saf., vol. 74, no. 2, pp. 53–61, 2009, doi: 10.1111/j.1750-3841.2008.01030.x.

F. Nazzaro, F. Fratianni, R. Coppola, A. Sada, and P. Orlando, “Fermentative ability of alginate-prebiotic encapsulated Lactobacillus acidophilus and survival under simulated gastrointestinal conditions,” J. Funct. Foods, vol. 1, no. 3, pp. 319–323, 2009, doi: 10.1016/j.jff.2009.02.001.

X. Y. Li, X. G. Chen, Z. W. Sun, H. J. Park, and D.-S. Cha, “Preparation of alginate chitosan/carboxymethyl chitosan complex microcapsules and application in Lactobacillus casei ATCC 393,” Carbohydr. Polym., vol. 83, no. 4, pp. 1479–1485, 2011, doi: 10.1016/j.carbpol.2010.09.053.

M. T. Cook, G. Tzortzis, D. Charalampopoulos, and V. V Khutoryanskiy, “Microencapsulation of a synbiotic into PLGA / alginate multiparticulate gels,” Int. J. Pharm., pp. 1–9, 2014, doi: 10.1016/j.ijpharm.2014.03.034.

S. Nualkaekul, D. Lenton, M. T. Cook, V. V. Khutoryanskiy, and D. Charalampopoulos, “Chitosan coated alginate beads for the survival of microencapsulated Lactobacillus plantarum in pomegranate juice,” Carbohydr. Polym., vol. 90, no. 3, pp. 1281–1287, 2012, doi: https://doi.org/10.1016/j.carbpol.2012.06.073.

K. G. H. Desai and H. J. Park, “Recent developments in microencapsulation of food ingredients Kashappa,” Dry. Technol., vol. 23, no. October, pp. 1361–1394, 2012, doi: https://doi.org/10.1081/DRT-200063478.

P. Muthukumarasamy, P. Wojtas, and R. A Holley, “M: Food microbiology and safety. Stability of Lactobacillus reuteri in different types of microcapsules,” J. Food Sci., vol. 71, no. 1, pp. 20–24, 2006, [Online]. Available: www.ift.org.

M. Xu, F. Gagné-Bourque, M.-J. Dumont, and S. Jabaji, “Encapsulation of Lactobacillus casei ATCC 393 cells and evaluation of their survival after freeze-drying , storage and under gastrointestinal conditions,” J. Food Eng., vol. 168, pp. 52–59, 2016, doi: 10.1016/j.jfoodeng.2015.07.021.

H. Gandomi, S. Abbaszadeh, A. Misaghi, S. Bokaie, and N. Noori, “Effect of chitosan-alginate encapsulation with inulin on survival of Lactobacillus rhamnosus GG during apple juice storage and under simulated gastrointestinal conditions,” LWT - Food Sci. Technol., vol. 69, pp. 365–371, 2016, doi: 10.1016/j.lwt.2016.01.064.

P. H. P. Prasanna and D. Charalampopoulos, “Encapsulation of Bifidobacterium longum in alginate-dairy matrices and survival in simulated gastrointestinal conditions , refrigeration , cow milk and goat milk,” Food Biosci., vol. 21, no. November 2017, pp. 72–79, 2018, doi: 10.1016/j.fbio.2017.12.002.

M. P. Silva, F. L. Tulini, M. M. Ribas, M. Penning, C. S. Fávaro-trindade, and D. Poncelet, “Microcapsules loaded with the probiotic Lactobacillus paracasei BGP-1 produced by co-extrusion technology using alginate / shellac as wall material : Characterization and evaluation of drying processes,” Food Res. Int. J., pp. 1–9, 2016, doi: https://doi.org/10.1016/j.foodres.2016.09.008.

P. Darjani, M. Hosseini, R. Kadkhodaee, and E. Milani, “Influence of prebiotic and coating materials on morphology and survival of a probiotic strain of Lactobacillus casei exposed to simulated gastrointestinal conditions,” LWT - Food Sci. Technol., vol. 73, pp. 162–167, 2016, doi: 10.1016/j.lwt.2016.05.032.

L.-E. Shi et al., “Encapsulation of Lactobacillus bulgaricus in carrageenan-locust bean gum coated milk microspheres with double layer structure,” LWT - Food Sci. Technol., vol. 54, no. 1, pp. 147–151, 2013, doi: https://doi.org/10.1016/j.lwt.2013.05.027.

C. Caren, G. Brusch, R. Michele, D. Soares, M. Antônio, and Z. Ayub, “Electrospraying microencapsulation of Lactobacillus plantarum enhances cell viability under refrigeration storage and simulated gastric and intestinal fluids,” J. Funct. Foods, vol. 24, pp. 316–326, 2016, doi: 10.1016/j.jff.2016.03.036.

M. Chen, W. Zheng, Q. Dong, Z. Li, and L. Shi, “Activity of encapsulated Lactobacillus bulgaricus in alginate-whey protein microspheres,” Braz. Arch. Biol. Technol., vol. 57, no. 5, pp. 736–741, 2014, doi: http://dx.doi.org/10.1590/S1516-8913201402377.

H. Zheng et al., “An improved pH-responsive carrier based on EDTA-Ca-alginate for oral delivery of Lactobacillus rhamnosus ATCC 53103,” Carbohydr. Polym., vol. 155, pp. 329–335, 2017, doi: 10.1016/j.carbpol.2016.08.096.

A. B. Shori, “Microencapsulation improved probiotics survival during gastric transit,” HAYATI J. Biosci., vol. 24, no. 1, pp. 1–5, 2017, doi: 10.1016/j.hjb.2016.12.008.

L. Chen, T. Yang, Y. Song, G. Shu, and H. Chen, “Effect of xanthan-chitosan-xanthan double layer encapsulation on survival of Bifidobacterium BB01 in simulated gastrointestinal conditions , bile salt solution and yogurt,” LWT - Food Sci. Technol., vol. 81, pp. 274–280, 2017, doi: 10.1016/j.lwt.2017.04.005.

M. P. Silva, F. L. Tulini, E. Martins, M. Penning, C. S. Fávaro-trindade, and D. Poncelet, “Comparison of extrusion and co-extrusion encapsulation techniques to protect Lactobacillus acidophilus LA3 in simulated gastrointestinal fl uids,” LWT - Food Sci. Technol., vol. 89, no. June 2017, pp. 392–399, 2018, doi: 10.1016/j.lwt.2017.11.008.

R. Sunaryanto, E. Martius, and B. Marwoto, “Uji kemampuan Lactobacillus casei sebagai agensia probiotik,” J. Bioteknol. Biosains Indones., vol. 1, no. 1, pp. 9–14, 2014, [Online]. Available: http://ejurnal.bppt.go.id/index.php/JBBI.

S. Cai, M. Zhao, Y. Fang, K. Nishinari, and G. O. Phillips, “Microencapsulation of Lactobacillus acidophilus CGMCC1 . 2686 via emulsification/internal gelation of alginate using Ca-EDTA and CaCO3 as calcium sources,” Food Hydrocoll., vol. 39, pp. 295–300, 2014, doi: 10.1016/j.foodhyd.2014.01.021.

M. A. K. Budiyanto, “Optimasi pengembangan kelembagaan industri pangan organik di jawa timur,” J. Tek. Ind., vol. 12, no. 2, pp. 169–176, 2011.

D. Sgouras et al., “In Vitro and In Vivo Inhibition of Helicobacter pylori by Lactobacillus casei Strain Shirota,” Appl. Environ. Microbiol., vol. 70, no. 1, pp. 518–526, 2004, doi: 10.1128/AEM.70.1.518-526.2004.

T. Heidebach, P. F. Rst, and U. Kulozik, “Microencapsulation of probiotic cells by means of rennet-gelation of milk proteins,” Food Hydrocoll., vol. 23, pp. 1670–1677, 2009, doi: 10.1016/j.foodhyd.2009.01.006.

T. Heidebach, P. F. Rst, and U. Kulozik, “Transglutaminase-induced caseinate gelation for the microencapsulation of probiotic cells,” Int. Dairy J. J., vol. 19, pp. 77–84, 2009, doi: 10.1016/j.idairyj.2008.08.003.

S. B. Doherty, M. A. Auty, C. Stanton, R. P. Ross, G. F. Fitzgerald, and A. Brodkorb, “Survival of entrapped Lactobacillus rhamnosus GG in whey protein micro-beads during simulated ex vivo gastro-intestinal transit,” Int. Dairy J., vol. 22, no. 1, pp. 31–43, 2012, doi: https://doi.org/10.1016/j.idairyj.2011.06.009.

K. Sultana, G. Godward, N. Reynolds, R. Arumugaswamy, P. Peiris, and K. Kailasapathy, “Encapsulation of probiotic bacteria with alginate-starch and evaluation of survival in simulated gastrointestinal conditions and in yoghurt,” Int. J. Food Microbiol., vol. 62, no. 1–2, pp. 47–55, 2000, doi: 10.1016/S0168-1605(00)00380-9.

A. K. Anal and H. Singh, “Recent advances in microencapsulation of probiotics for industrial applications and targeted delivery,” Trends Food Sci. Technol., vol. 18, pp. 240–251, 2007, doi: 10.1016/j.tifs.2007.01.004.

R. R. Mokarram, S. A. Mortazavi, M. B. H. Najafi, and F. Shahidi, “The influence of multi-stage alginate coating on survivability of potential probiotic bacteria in simulated gastric and intestinal juice q,” Food Res. Int., vol. 42, no. 8, pp. 1040–1045, 2009, doi: 10.1016/j.foodres.2009.04.023.

M. Chávarri, I. Marañón, R. Ares, F. C. Ibáñez, F. Marzo, and C. Villarán, “Microencapsulation of a probiotic and prebiotic in alginate-chitosan capsules improves survival in simulated gastro-intestinal conditions,” Int. J. Food Microbiol., vol. 142, no. 1–2, pp. 185–189, 2010, doi: 10.1016/j.ijfoodmicro.2010.06.022.

M. T. Cook, G. Tzortzis, D. Charalampopoulos, and V. V Khutoryanskiy, “Production and evaluation of dry alginate-chitosan microcapsules as an enteric delivery vehicle for probiotic bacteria,” Biomacromolecules, pp. 2834–2840, 2011, doi: https//doi.org/10.1021/bm200576h.

M. T. Cook, G. Tzortzis, V. V Khutoryanskiy, and D. Charalampopoulos, “chitosan – alginate for the improved survival and administration †,” J. Mater. Chem. B, vol. 1, pp. 52–60, 2013, doi: https://doi.org/10.1039/c2tb00126h.

W. Krasaekoopt, B. Bhandari, and H. Deeth, “Evaluation of encapsulation techniques of probiotics for yoghurt,” Int. Dairy J., vol. 13, no. 1, pp. 3–13, 2003, doi: 10.1016/S0958-6946(02)00155-3.

L. de Souza Simões, D. A. Madalena, A. C. Pinheiro, J. A. Teixeira, A. A. Vicente, and Ó. L. Ramos, “Micro- and nano bio-based delivery systems for food applications: In vitro behavior,” Adv. Colloid Interface Sci., vol. 243, pp. 23–45, 2017, doi: 10.1016/j.cis.2017.02.010.

W. K. DING and N. P. SHAH, “Effect of various encapsulating materials on the stability of probiotic bacteria,” J. Food Sci., vol. 74, no. 2, 2009, doi: 10.1111/j.1750-3841.2009.01067.x.

M. José, F. Lara-villoslada, M. Adol, and M. Encarnación, “Microencapsulation of bacteria : A review of different technologies and their impact on the probiotic effects,” Innov. Food Sci. Emerg. Technol., vol. 27, pp. 15–25, 2015, doi: 10.1016/j.ifset.2014.09.010.

P. Allan-Wojtas, L. T. Hansen, and A. T. Paulson, “Microstructural studies of probiotic bacteria-loaded alginate microcapsules using standard electron microscopy techniques and anhydrous fixation $,” LWT, vol. 41, pp. 101–108, 2008, doi: 10.1016/j.lwt.2007.02.003.

J. Burgain, C. Gaiani, M. Linder, and J. Scher, “Encapsulation of probiotic living cells: From laboratory scale to industrial applications,” J. Food Eng., vol. 104, no. 4, pp. 467–483, 2011, doi: 10.1016/j.jfoodeng.2010.12.031.

S. Prakash and M. L. Jones, “Artificial cell therapy : New strategies for the therapeutic delivery of live bacteria,” J. Biomed. Biotechnol., vol. 1, pp. 44–56, 2005, doi: 10.1155/JBB.2005.44.

L. T. Hansen, P.M.Allan-Wojtas, Y.-L. Jin, and A. T. Paulson, “Survival of Ca-alginate microencapsulated Bifidobacterium spp. in milk and simulated gastrointestinal conditions,” Food Microbiol., vol. 19, pp. 35–45, 2002, doi: 10.1006/fmic.2001.0452.

C. E. Chitnis and D. E. Ohmant, “Cloning of Pseudomonas aeruginosa algG , which controls alginate structure G-G-G G block,” J. Bacteriol., vol. 172, no. 6, pp. 2894–2900, 1990.

N. Nwe, T. Furuike, and H. Tamura, “The mechanical and biological properties of chitosan scaffolds for tissue regeneration templates are significantly enhanced by chitosan from Gongronella butleri,” Materials (Basel)., vol. 2, no. 2, pp. 374–398, 2009, doi: 10.3390/ma2020374.

F. Liaqat and R. Eltem, “Chitooligosaccharides and their biological activities: A comprehensive review,” Carbohydr. Polym., vol. 184, no. June 2017, pp. 243–259, 2018, doi: 10.1016/j.carbpol.2017.12.067.

D. E. Wright, “Toxins produced by fungi,” Annu. Rev. Microbiol., vol. 22, no. 1, pp. 269–282, 1968, doi: 10.1146/annurev.mi.22.100168.001413.

A. R. Logesh, K. A. Thillaimaharani, K. Sharmila, M. Kalaiselvam, and S. M. Raffi, “Production of chitosan from endolichenic fungi isolated from mangrove environment and its antagonistic activity,” Asian Pac. J. Trop. Biomed., vol. 2, no. 2, pp. 140–143, 2012, doi: 10.1016/S2221-1691(11)60208-6.

J. G. Fernandez and D. E. Ingber, “Manufacturing of large-scale functional Objects using biodegradable chitosan bioplastic,” macromolecular, vol. 299, pp. 932–938, 2014, doi: https://doi.org/10.1002/mame.201300426.

T. Barclay, M. Ginic-markovic, P. Cooper, and N. Petrovsky, “Inulin - a versatile polysaccharide with multiple pharmaceutical and food chemical uses,” J. Excipients Food Chem., vol. 1, no. 3, pp. 27–50, 2010, [Online]. Available: nikolai.petrovsky@flinders.edu.au.

R. M. Lucinda-silva, H. Regina, N. Salgado, and R. Cesar, “Alginate – chitosan systems : In vitro controlled release of triamcinolone and in vivo gastrointestinal transit,” Carbohydr. Polym., vol. 81, pp. 260–268, 2010, doi: 10.1016/j.carbpol.2010.02.016.

D. Bosscher, J. Van Loo, and A. Franck, “Inulin and oligofructose as prebiotics in the prevention of intestinal infections and diseases,” Nutr. Res. Rev., vol. 19, pp. 216–226, 2006, doi: 10.1017/NRR2006127.




How to Cite

Djaenudin, Saepudin, E. ., & Nasir, M. . (2021). Microencapsulation of Probiotic <em>Lactobacillus casei </em>based on Alginate and Chitosan Materials . Asian Journal of Applied Sciences, 9(2). https://doi.org/10.24203/ajas.v9i2.6564