Probiotics and Antimicrobial Proteins

, Volume 11, Issue 4, pp 1340–1347 | Cite as

Survival and Goat Milk Acidifying Activity of Lactobacillus rhamnosus GG Encapsulated with Agave Fructans in a Buttermilk Protein Matrix

  • Octavio Alvarado-Reveles
  • Silvia Fernández-Michel
  • Rafael Jiménez-Flores
  • Cristina Cueto-Wong
  • Luz Vázquez-Moreno
  • Gabriela Ramos-Clamont MontfortEmail author


Lactobacillus rhamnosus GG (L. rhamnosus GG) cells were encapsulated in buttermilk proteins by spray drying, alone (E), or with Agave tequilana fructans (CEF). Buttermilk proteins acted as a thermo-protector for the probiotic cells undergoing the spray-dried process. The addition of Agave fructans in CEF microcapsules significantly enhanced storage stability and survival to in vitro simulated gastrointestinal conditions, compared to E capsules. After 14 days storage at − 20 °C, the number of living cells in CEF microcapsules was in the order of 7.7 log CFU • mL−1 and the survivability in simulated gastrointestinal environment was 73.23%. Spray-dried microparticles were cultured in goat milk to study biomass production. Agave fructans offered a favorable microenvironment and better growth substrate. The population of CEF viable cells reached 1.08 ± 0.02 × 1010 CFU • mL−1 after 18 h of fermentation. In contrast, the population of E viable cells were 3.0 ± 0.01 × 109 CFU • mL−1. The generation time of CEF, L. rhamnosus GG was 15% faster than E, L. rhamnosus GG. Encapsulation with buttermilk proteins in the presence of Agave fructans by spray drying could be suitable for preservation of probiotic powders and may be for a more effective application of probiotics in goat dairy products.


Probiotics Encapsulation Buttermilk proteins Agave fructans 



We are grateful to the National Council of Science and Technology of Mexico, CONACyT, for the financial support for this research, under project CB169358, as well as for the scholarship awarded for MSc. studies. The authors are thankful to Ana María Domínguez Vergara and Hayde Gonzalez Carrillo for their technical assistance in this work.


This study was supported by the National Council of Science and Technology of Mexico, CONACyT under project CB169358.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflicts of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. 1.
    Meira QGS, Magnani M, de Medeiros Júnior FC, Queiroga RCRE, Madruga MS, Gullón B, Gomes AMP, Pintado MME, de Souza EL (2015) Effects of added Lactobacillus acidophilus and Bifidobacterium lactis probiotics on the quality characteristics of goat ricotta and their survival under simulated gastrointestinal conditions. Food Res Int 76:828–838. CrossRefPubMedGoogle Scholar
  2. 2.
    Raynal-Ljutovac K, Lagriffoul G, Paccard P, Guillet I, Chilliard Y (2008) Composition of goat and sheep milk products: an update. Small Rumin Res 79:57–72. CrossRefGoogle Scholar
  3. 3.
    Salva S, Nuñez M, Villena J, Ramón A, Font G, Alvarez S (2011) Development of a fermented goats’ milk containing Lactobacillus rhamnosus: in vivo study of health benefits. J Sci Food Agric 91:2355–2362. CrossRefPubMedGoogle Scholar
  4. 4.
    Burgain J, Gaiani C, Linder M, Scher J (2011) Encapsulation of probiotic living cells: from laboratory scale to industrial applications. J Food Eng 104:467–483. CrossRefGoogle Scholar
  5. 5.
    Gomez E, Tuohy KM, Gibson GR, Klinder A, Costabile A (2010) In vitro evaluation of the fermentation properties and potential prebiotic activity of Agave fructans. J Appl Microbiol 108:2114–2121. CrossRefPubMedGoogle Scholar
  6. 6.
    Anal AK, Singh H (2007) Recent advances in microencapsulation of probiotics for industrial applications and targeted delivery. Trends Food Sci Technol 18:240–251. CrossRefGoogle Scholar
  7. 7.
    Ying D, Schwander S, Weerakkody R, Sanguansri L, Gantenbein-Demarchi C, Augustin MA (2013) Microencapsulated Lactobacillus rhamnosus GG in whey protein and resistant starch matrices: probiotic survival in fruit juice. J Funct Foods 5:98–105. CrossRefGoogle Scholar
  8. 8.
    Heidebach T, Först P, Kulozik U (2009) Transglutaminase-induced caseinate gelation for the microencapsulation of probiotic cells. Int Dairy J 19:77–84. CrossRefGoogle Scholar
  9. 9.
    Crittenden R, Weerakkody R, Sanguansri L, Augustin M (2006) Synbiotic microcapsules that enhance microbial viability during nonrefrigerated storage and gastrointestinal transit. Appl Environ Microbiol 72:2280–2282. CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Livney YD (2010) Milk proteins as vehicles for bioactives. Curr Opinion Colloid Interface Sci 15:73–83. CrossRefGoogle Scholar
  11. 11.
    Figueroa Valenzuela C, Meda Gutiérrez F, Janacua Vidales H (2004) Manual de Buenas Prácticas en Producción de Leche Caprina. Accessed 10 Jan 2017
  12. 12.
    Fritzen-Freire CB, Prudêncio ES, Amboni RDMC, Pinto SS, Negrão-Murakami AN, Murakami FS (2012) Microencapsulation of bifidobacteria by spray drying in the presence of prebiotics. Food Res Int 45:306–312. CrossRefGoogle Scholar
  13. 13.
    Doleyres Y, Fliss I, Lacroix C (2004) Increased stress tolerance of Bifidobacterium longum and Lactococcus lactis produced during continuous mixed-strain immobilized-cell fermentation. J Appl Microbiol 97:527–539. CrossRefPubMedGoogle Scholar
  14. 14.
    Oliveira RPDS, Perego P, de Oliveira MN, Converti A (2012) Effect of inulin on the growth and metabolism of a probiotic strain of Lactobacillus rhamnosus in co-culture with Streptococcus thermophilus. LWT Food Sci Technol 47:358–363. CrossRefGoogle Scholar
  15. 15.
    Kristo E, Biliaderis CG, Tzanetakis N (2003) Modelling of rheological, microbiological and acidification properties of a fermented milk product containing a probiotic strain of Lactobacillus paracasei. Int Dairy J 13:517–528. CrossRefGoogle Scholar
  16. 16.
    Horwitz W, Latimer GW (2005) Official methods of analysis of AOAC international. AOAC International, GaithersburgGoogle Scholar
  17. 17.
    Heidebach T, Forst P, Kulozik U (2012) Microencapsulation of probiotic cells for food applications. Crit Rev Food Sci Nutr 52:291–311. CrossRefPubMedGoogle Scholar
  18. 18.
    Mokarram RR, Mortazavi SA, Najafi MBH, Shahidi F (2009) The influence of multi stage alginate coating on survivability of potential probiotic bacteria in simulated gastric and intestinal juice. Food Res Int 42:1040–1045. CrossRefGoogle Scholar
  19. 19.
    Ding WK, Shah NP (2009) Effect of various encapsulating materials on the stability of probiotic bacteria. J Food Sci 74:M100–M107. CrossRefPubMedGoogle Scholar
  20. 20.
    Saénz C, Tapia S, Chávez J, Robert P (2009) Microencapsulation by spray drying of bioactive compounds from cactus pear (Opuntia ficus-indica). Food Chem 114:616–622. CrossRefGoogle Scholar
  21. 21.
    Engelen L, Van der Bilt A, Schipper M, Bosman F (2005) Oral size perception of particles: effect of size, type, viscosity and method. J Texture Stud 36:373–386. CrossRefGoogle Scholar
  22. 22.
    Imai E, Hatae K, Shimada A (1995) Oral perception of grittiness: effect of particle size and concentration of the dispersed particles and the dispersion medium. J Texture Stud 26:561–576. CrossRefGoogle Scholar
  23. 23.
    Sarkar A, Kanti F, Gulotta A, Murray BS, Zhang S (2017) Aqueous lubrication, structure and rheological properties of whey protein microgel particles. Langmuir 33:14699–14708. CrossRefPubMedGoogle Scholar
  24. 24.
    Burgain J, Gaiani C, Cailliez-Grimal C, Jeandel C, Scher J (2013) Encapsulation of Lactobacillus rhamnosus GG in microparticles: influence of casein to whey protein ratio on bacterial survival during digestion. Innovative Food Sci Emerg Technol 19:233–242. CrossRefGoogle Scholar
  25. 25.
    Morin P, Pouliot Y, Jiménez-Flores R (2006) A comparative study of the fractionation of regular buttermilk and whey buttermilk by microfiltration. J Food Eng 77:521–528. CrossRefGoogle Scholar
  26. 26.
    Capela P, Hay TKC, Shah NP (2006) Effect of cryoprotectants, prebiotics and microencapsulation on survival of probiotic organisms in yoghurt and freeze-dried yoghurt. Food Res Int 39:203–211. CrossRefGoogle Scholar
  27. 27.
    Ying D, Sun J, Sanguansri L, Weerakkody R, Augustin MA (2012) Enhanced survival of spray-dried microencapsulated Lactobacillus rhamnosus GG in the presence of glucose. J Food Eng 109:597–602. CrossRefGoogle Scholar
  28. 28.
    Espinosa-Andrews H, Rodríguez-Rodríguez R (2018) Water state diagram and thermal properties of fructans powders. J Therm Anal Calorim 132:197–204. CrossRefGoogle Scholar
  29. 29.
    Schaller-Povolny LA, Smith DE, Labuza TP (2000) Effect of water content and molecular weight on the moisture isotherms and glass transition properties of inulin. Int J Food Prop 3:173–192. CrossRefGoogle Scholar
  30. 30.
    Ying DY, Phoon MC, Sanguansri L, Weerakkody R, Burgar I, Augustin MA (2010) Microencapsulated Lactobacillus rhamnosus GG powders: relationship of powder physical properties to probiotic survival during storage. J Food Sci 75:E588–E595. CrossRefPubMedGoogle Scholar
  31. 31.
    Dianawati D, Mishra V, Shah NP (2016) Survival of microencapsulated probiotic bacteria after processing and during storage: a review. Crit Rev Food Sci Nutr 56:1685–1716. CrossRefPubMedGoogle Scholar
  32. 32.
    Rajam R, Anandharamakrishnan C (2015) Microencapsulation of Lactobacillus plantarum (MTCC 5422) with fructooligosaccharides as wall material by spray drying. LWT Food Sci Technol 60:773–780. CrossRefGoogle Scholar
  33. 33.
    Sosa N, Gerbino E, Golowczyc MA, Schebor C, Gómez-Zavaglia A, Tymczyszyn EE (2016) Effect of galactooligosaccharides: maltodextrin matrices on the recovery of Lactobacillus plantarum after spray-drying. Front Microbiol 7:584. CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Pinto SS, Verruck S, Vieira CRW, Prudêncio ES, Amante ER, Amboni RDMC (2015) Influence of microencapsulation with sweet whey and prebiotics on the survival of Bifidobacterium-BB-12 under simulated gastrointestinal conditions and heat treatments. LWT Food Sci Technol 64:1004–1009. CrossRefGoogle Scholar
  35. 35.
    Park YW, Juárez M, Ramos M, Haenlein GFW (2007) Physico-chemical characteristics of goat and sheep milk. Small Rumin Res 68:88–113. CrossRefGoogle Scholar
  36. 36.
    Kongo JM, Gomes AM, Malcata FX (2006) Manufacturing of fermented goat milk with a mixed starter culture of Bifidobacterium animalis and Lactobacillus acidophilus in a controlled bioreactor. Lett Appl Microbiol 42:595–599. CrossRefPubMedGoogle Scholar
  37. 37.
    Gbassi GK, Vandamme T (2012) Probiotic encapsulation technology: from microencapsulation to release into the gut. Pharmaceutics 4:149–163. CrossRefPubMedPubMedCentralGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Facultad de Ciencias BiológicasUniversidad Autónoma de CoahuilaTorreónMexico
  2. 2.Dairy Products Technology CenterCalifornia Polytechnic State UniversitySan Luis ObispoUSA
  3. 3.Centro de Investigación en Alimentación y DesarrolloA.C. Coordinación de Ciencia de los AlimentosHermosilloMexico

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