Applied Microbiology and Biotechnology

, Volume 38, Issue 1, pp 39–45 | Cite as

Production of alginate beads by emulsification/internal gelation. I. Methodology

  • D. Poncelet
  • R. Lencki
  • C. Beaulieu
  • J. P. Halle
  • R. J. Neufeld
  • A. Fournier


Small diameter alginate beads (microspheres) were formed via internal gelation of alginate solution emulsified within vegetable oil. Gelation was initiated by addition of an oil-soluble acid thereby reducing the pH of the alginate solution and releasing soluble Ca2+ from the citrate complex. Smooth, spherical, micron-sized beads were formed. The mean diameter ranged from 200 to 1000 μm, controlled by the reactor impeller design and rotational speed. The technique has potential for large-scale and continuous applications in immobilization.


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  1. Audet P, Lacroix C (1989) Two-phase dispersion process for the production of biopolymer gel beads: effect of various parameters on bead size and their distribution. Process Biochem 24:217–225Google Scholar
  2. Burns MA, Kvesitadze GI, Graves DJ (1985) Dried calcium alginate/magnetite spheres: a new support for chromatographic separations and enzyme immobilization. Biotechnol Bioeng 27:137–145Google Scholar
  3. Calabrese RV, Wang CY, Bryner NP (1986) Drop breakup in turbulent stirred-tank contactors; part 3: correlations for mean size and drop size distribution. AIChE J 32:677–681Google Scholar
  4. Canon K (1984) Electrostatic image development toners. Japanese patent no. 59 170.853Google Scholar
  5. Champagne CP, Gaudy C, Poncelet D, Neufeld RJ (1991) Lactococcus lactis cell release from calcium alginate beads. Appl Environ Microbiol 58:1429–1434Google Scholar
  6. Chang TMS, MacIntosh FC, Mason SG (1966) Semipermeable aqueous microcapsules. I. Preparation and properties. Can J Physiol Pharmacol 44:115–128Google Scholar
  7. Dabora EK (1967) Production of monodisperse sprays. Rev Sci Instrum 38:502–506Google Scholar
  8. Grace HP (1982) Dispersion phenomena in high viscosity immiscible fluid systems and application of static mixers as dispersion devices in such systems. Chem Eng Commun 14:225–277Google Scholar
  9. Haas PA (1987) Turbulent dispersion of aqueous drops in organic liquids. AIChE J 33:987–995Google Scholar
  10. Hinze JO (1955) Fundamentals of the hydrodynamic mechanism of splitting in dispersion processes. AIChE J 1:289Google Scholar
  11. Hommel M, Sun AM, Goosen MFA (1988) Droplets generation. Canadian patent no. 1241598Google Scholar
  12. Hulst AC, Tramper J, Brodelius P, Eijkenboom LJC, Luyben CAM (1985) Immobilised plant cells: respiration and oxygen transfer. J Chem Technol Biotechnol 35B:198–204Google Scholar
  13. Kennedy JF, Cabral JMS (1983) Immobilized enzymes in solid phase biochemistry. Chem Anal 66:253–392Google Scholar
  14. Kierstan M, Bucke C (1977) The immobilization of microbial cells, subcellular organelles, and enzymes in calcium alginate gels. Biotechnol Bioeng 19:387–397Google Scholar
  15. Lacroix C, Paquin C, Arnaud J-P (1990) Batch fermentation with entrapped growing cells of Lactobacillus casei; optimization of the rheological properties of the entrapment gel matrix. Appl Microbiol Biotechnol 32:403–408Google Scholar
  16. Lencki RWJ, Neufeld RJ, Spinney T (1989) Microspheres and method of producing same. US patent no. 4822534Google Scholar
  17. Lim F (1983) Microcapsules containing viable tissue cells. US patent no. 4391909Google Scholar
  18. Lim F, Sun AM (1980) Microencapsulated islets as bioartificial endocrine pancreas. Science 210:908–910Google Scholar
  19. Miyawaki O, Nakamura K, Yano T (1980) Permeability and molecular sieving characteristics of nylon microcapsule membrane. Agric Biol Chem 44:2865–2870Google Scholar
  20. Neufeld RJ, Peleg Y, Rokem JS, Pines O, Goldberg I (1991) l-Malic acid formation by immobilized Saccaromyces cerevisiae amplified for fumarase. Enzyme Microb Technol 13:991–996Google Scholar
  21. Nilsson K, Scheider W, Katinger HWD, Mosbach K (1986) Production of monoclonal antibodies by agarose-entrapped hybridoma cells. Methods Enzymol 121:352–360Google Scholar
  22. Oldshue JY (1983) Fluid mixing technology. McGraw-Hill, New York, pp 12–14Google Scholar
  23. Pelaez C, Karel M (1981) Improved method for preparation of fruit-simulating alginate gels. J Food Process Preserv 5:63–81Google Scholar
  24. Poncelet D, Poncelet De Smet B, Beaulieu C, Neufeld RJ (1992) Scale-up of gel bead and microcapsule production in cell immobilization. In: Goosen MFA (eds) Fundamentals of animal cell encapsulation and immobilization. CRC Press, Boca Raton, Fla., in pressGoogle Scholar
  25. Poncelet De Smet B, Poncelet D, Neufeld RJ (1989) Control of mean diameter and size distribution during formulation of microcapsules with cellulose nitrate membranes. Enzyme Microb Technol 11:29–37Google Scholar
  26. Poncelet De Smet B, Poncelet D, Neufeld RJ (1990) Preparation of hemolysate-filled hexamethylene sebacamide microcapsules with controlled diameter. Can J Chem Eng 68:443–448Google Scholar
  27. Provost H, Diovies C, Rousseau E (1985) Continuous production with Lactobacillus bulgaricus and Streptococcus thermophilus entrapped in calcium alginate. Biotechnol Lett 7:247–252Google Scholar
  28. Q. P. Corporation (1985) Production of fish egg-like spherical food material. Japanese patent no. JP 60 83.570Google Scholar
  29. Redenbaugh K, Paasch BD, Nichol JW, Kossler ME, Viss PR, Walker KA (1986) Somatic seeds: encapsulation of asexual plant embryos. Bio/Technology 4:797–801Google Scholar
  30. Rounsley RR (1983) Oil dispersion with a turbine mixer. AIChE J 29:597–603Google Scholar
  31. Shiotani T, Yamane T (1981) A horizontal packed-bed bioreactor to reduce carbon dioxide gas holdup in the continuous production of ethanol in immobilized yeast cells. Eur J Appl Microbiol Biotechnol 13:96–101Google Scholar
  32. Smidsrod O, Skjak-Braek G (1990) Alginate as immobilization matrix for cells. Trends Biotechnol 8:71–78Google Scholar
  33. Su H, Bajpai R, Preckshot GW (1989) Characterization of alginate beads formed by a two fluid annular atomizer. Appl Biochem Biotechnol 20/21:561–569Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • D. Poncelet
    • 1
  • R. Lencki
    • 2
  • C. Beaulieu
    • 1
  • J. P. Halle
    • 3
  • R. J. Neufeld
    • 2
  • A. Fournier
    • 1
  1. 1.INRS-SantéUniversité du QuébecQuébecCanada
  2. 2.Department of Chemical EngineeringMcGill UniversityMontréal, QuébecCanada
  3. 3.Maisonneuve Rosemont HospitalMontréal, QuébecCanada
  4. 4.Department of Food ScienceUniversity of GuelphGuelphCanada

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