Abstract
Cyclodextrin-glycosyl-transferase (EC2.4.1.19), produced by Wacker (Munich, Germany), was purified by biospecific affinity chromatography with β-cyclodextrin (β-CD) as ligand, and immobilized into controlled pore silica particles (0.42 mm). This immobilized enzyme (IE) had 4.7 mg of protein/g of support and a specific activity of 8.6 μmol of β-CD/(min·gIF) at 50°C, pH 8.0. It was used in a fluidized-bed reactor (FBR) at the same conditions for producing cyclodextrins (CDs) with 10% (w/v) maltodextrin solution as substrate. Bed expansion was modeled by the Richardson and Zaki equation, giving a good fit in two distin ctranges of bed porosities. The minimum fluidization velocity was 0.045 cm/s, the bed expansion coefficient was 3.98, and the particle terminal velocity was 2.4 cm/s. The FBR achieved high productivity, reaching in only 4 min of residence time the same amount of CDs normally achieved in a batch reactor with free enzyme after 24h of reaction, namely, 10.4 mM β-CD and 2.3 mM γ-CD.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Referneces
Szejtli, J. (1990), in Carbohydrate Polymers, vol. 12, Elsevier Science, pp. 375–392.
Frömming, K. H. and Szejtli, J. (1994), Cyclodextrins in Pharmacy, Davies, J. E. D., ed., Kluwer Academic, Dordrecht, Netherlands, pp. 1–81.
Szejtli, J. (1988), Cyclodextrin Technology, Davies, J. E. D., ed., Kluwer Academic, Dordrecht, Netherlands, pp. 79–185.
Bekers, O., Uijtendaal, E. V., Beijnen, J. H., Bult, A., and Underberg, G. W. J. M. (1991), Drug Dev. Ind. Pharm. 17, 1503–1549.
Korolkovas, A. (1991), ENLACE Farmalab 2, 6–15.
Moffatt, J. R. (1992), US patent 5,108,505.
Horikoshi, K., Nakamura, N., Matsuzawa, N., and Yamamoto, M. (1982), in Proceedings of the First International Symposium on Cyclodextrins, Szejtli, J., ed., D. Reidel Publishing, Dordrecht, Netherlands, pp. 25–39.
Lima, H. O. S., de Moraes, F. F., and Zanin, G. M. (1998), Appl. Biochem. Biotechnol. 70–72, 789–804.
László, E., Banky, B., Seres, G., and Szejtli J. (1981), Starch 33, 281–283.
Berna, P., de Moraes, F. F., Barbotin, J. N., Thomas, D., and Vijayalaksmi, M. A. (1996), in Advances in Molecular and Cell Biology vol. 15B, Bittar, E. E., ed., JAI Press, Greenwich, CT, pp. 521–535.
Pedersen, S. (1993), in Industrial Application of Immobilized Biocatalysts, Tanaka, A., Tosa, T., and Kobayashi, T., eds., Marcel Dekker, New York, pp. 185–208.
Tramper, J. (1994), in Applied Biocatalysis, Cabral, J. M. S., Best, D., Boross, L., and Tramper, J., eds., Harwood Academic, Chur, Switzerland, pp. 1–45.
Zanin, G. M., Kambara, L. M., Calsavara, L. P. V., and de Moraes, F. F. (1994), Appl. Biochem. Biotechnol. 45–46, 627–640.
Schmid, G., Engelbrecht, A., and Schmid, D. (1988), in Proceedings of the Fourth International Symposium on Cyclodextrins, Huber, O. and Szejtli, J., eds., Kluwer Academic, Dordrecht, Netherlands, pp. 71–76.
Weetall, H. H. (1993), Appl. Biochem. Biotechnol. 41, 157–187.
Catry, S. and de Moraes, F. F. (1990), Research Report, Laboratoire de Technologie Enzymatique, Université de Technologie de Compiègne, Compiègne, France.
Richardson, J. H. and Zaki, W. N. (1954), Trans. Inst. Chem. Eng. 32, 35–53.
Vikmon, M. (1982), in Proceedings of the First International Symposium on Cyclodextrins, Szejtli, J., ed., D. Reidel Publishing, Dordrecht, Netherlands, pp. 69–74.
Hamon, V. and de Moraes, F. F. (1990), Research Report. Laboratoire de Technologie Enzymatique, Université de Technologie de Compiègne, Compiègne, France.
Kato, T. and Horikoshi, D. (1984), Analyt. Chem. 56, 1738–1740.
Bradford, M. M. (1976), Analyt. Biochem. 72, 248–254.
de Moraes, F. F., Catry, S., Delbourg, M. F., Thomas, D., and Barbotin J. N. (1990), in The Industrial Applications of Natural, Modified and Artificial Enzymes, Carra, S., Ciardelli, F., and Houben, J. L., eds., Lito Felíci, Pisa, Italy, September 23–29, pp. 183, 184.
Crump, S. P. and Rozzell, J. D. (1988), in Proceedings of the Fourth International Symposium on Cyclodextrins, Huber, O. and Szejtli, J., eds., Kluwer Academic, Dordrecht, Netherlands, pp. 47–53.
Riba, J. P. and Courdec, J. P. (1977), Can. J. Chem. Eng. 55, 118–121.
Kunii, D. and Levenspiel, O. (1969), Fluidization Engineering, John Wiley & Sons, New York.
Nakamura, N. and Horikoshi, K., (1977), Biotechnol. Bioeng. 19, 87–99.
Steighardt, J. and Kleine, R. (1993), Appl. Microbiol. Biotechnol. 39, 63–68.
Su, C. S. and Yang, C. P. (1990), J. Chem. Technol. Biotechnol. 48, 313–323.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Tardioli, P.W., Zanin, G.M. & de Moraes, F.F. Production of cyclodextrins in a fluidized-bed reactor using cyclodextrin-glycosyl-transferase. Appl Biochem Biotechnol 84, 1003–1019 (2000). https://doi.org/10.1385/ABAB:84-86:1-9:1003
Issue Date:
DOI: https://doi.org/10.1385/ABAB:84-86:1-9:1003