Abstract
Intracellular frustosyl transferase was purified fromAureobasidium pullulans C-23 by ethanol fractionation, CM-Sephadex chromatography and preparative disc gel electrophoresis. It was shown to be homogeneous on disc polyacrylamide gel electrophoresis, with a molecular size of 190kDa. The pI value of the enzyme was about 3.7. The enzyme has aK m value of 0.43 mM for sucrose and was optimally active at pH 5.0 and 60°C. The enzyme was stable from pH 2.5 to 12. It was almost completely inhibited by 5mM Hg2+ but was not significantly affected by other cations. The transferase was inactivated by treatment with the tryptophan-specific reagentN-bromosuccinimide and the tyrosine-specific reagent, I2, suggesting that tryptophan and tyrosine residues are probably located at or near the active site of the enzyme.
Similar content being viewed by others
References
Bealing, F.J. 1953 Mould ‘Glucosaccharase’: a fructosidase.Biochemical Journal 55, 93–101.
Cho, W.-T. & Lim, J.-Y. 1990 Optimum culture conditions for production of fructosyl transferase byAureobasidium pullulans C-23.Korean Journal of Applied Microbiology and Biotechnology 18, 417–422.
Cho, W.-T., Lim, J.-Y. & Lee, S.-S. 1990 Isolation and identification of the black yeast producing fructosyl transferase.Korean Journal of Mycology 18, 20–25.
Dickerson, A.G. 1972 A β-D-fructofuranosidase fromClaviceps purpurea.Biochemical Journal 129, 263–272.
Edelman, J. 1954 Transfer reactions catalysed by some sucrase preparations.Biochemical Journal 57, 22–33.
Edelman, J. & Jefford, T.G. 1966 The metabolism of fructose polymers in plants.Biochemical Journal 98, 787–794.
Gupta, A.K. & Bhatia, I.S. 1980 Glucofructosan biosynthesis inFusarium oxysporum.Phystochemistry 19, 2557–2563.
Gupta, A.K. & Bhatia I.S. 1982 Glucofructosan biosynthesis inFusarium oxysporum.Phytochemistry 21, 1249–1253.
Hankin, L. & McIntyre, J.L. 1977 Production of kestose (fructosyl sucroses) byPhytophthora parasitica var.Nicotianae.Applied and Environmental Microbiology 33, 522–524.
Lee, K.-J., Cho, W.-T., Chang, Y.-S. & Lim, J.-Y. 1991 Purification and immobilization of extracellular fructosyl-transferase fromAureobasidium pullulans C-23.Genetic Engineering Research 5, 27–38.
Oku, T., Tokunaga, T. & Hosoya, N. 1984 Non-digestibility of a new sweetner, “Neosugar”, in the rat.Joural of Nutrition 114, 1574–1581.
Pazur, J.H. 1952 Transfructosidation reactions of an enzyme ofAspergillus oryzae.Journal of Biological Chemistry 199, 217–225.
Robert, J.H. & Darbyshire, B. 1980 Fructan synthesis in onion.Phytochemistry 19, 1017–1020.
Shiomi, N., Yamada, J. & Izawa, M. 1976 Isolation and identification of fructo-oligosaccharides in roots of asparagus.Agricultural and Biological Chemistry 40, 567–575.
Shiomi, N., Yamada, J. & Izawa, M. 1979 Synthesis of several fructo-oligosaccharides by asparagus fructosyl transferases.Agricultural and Biological Chemistry 43, 2233–2244.
Smith, J.A., Grove, D., Luenser, S.J. & Park, L.G. 1982 Process for the production of fructose transferase enzyme. United States Patent4, 309,505.
Wenham, D.G., Hennessey, T.D. & Cole, J.A. 1979 Regulation of glucosyl- and fructosyl transferase synthesis by continuous cultures ofStreptococcus mutans.Journal of General Microbiology 114, 117–124.
Yun, J.W., Jung, K.H., Oh, J.W. & Lee, J.H. 1989 Semibatch production of fructo-oligosaccharides from sucrose by immobilized cells ofAureobasidium pullulans.Journal of the Institute of Biotechnology 1, 16–21.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lee, K.J., Choi, J.D. & Lim, J.Y. Purification and properties of intracellular fructosyl transferase fromAureobasidium pullulans . World J Microbiol Biotechnol 8, 411–415 (1992). https://doi.org/10.1007/BF01198756
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01198756