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Purification and Characterization of Aspergillus terreus α-Galactosidases and Their Use for Hydrolysis of Soymilk Oligosaccharides

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Abstract

α-Galactosidases has the potential to hydrolyze α-1-6 linkages in raffinose family oligosaccharides (RFO). Aspergillus terreus cells cultivated on wheat bran produced three extracellular forms of α-galactosidases (E1, E2, and E3). E1 and E2 α-galactosidases presented maximal activities at pH 5, while E3 α-galactosidase was more active at pH 5.5. The E1 and E2 enzymes showed stability for 6 h at pH 4–7. Maximal activities were determined at 60, 55, and 50°C, for E1, E2, and E3 α-galactosidase, respectively. E2 α-galactosidase retained 90% of its initial activity after 70 h at 50°C. The enzymes hydrolyzed ρNPGal, melibiose, raffinose and stachyose, and E1 and E2 enzymes were able to hydrolyze guar gum and locust bean gum substrates. E1 and E3 α-galactosidases were completely inhibited by Hg2+, Ag+, and Cu2+. The treatment of RFO present in soy milk with the enzymes showed that E1 α-galactosidase reduced the stachyose content to zero after 12 h of reaction, while E2 promoted total hydrolysis of raffinose. The complete removal of the oligosaccharides in soy milk could be reached by synergistic action of both enzymes

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References

  1. Murata, T., & Usui, T. (1997). Bioscience, Biotechnology, and Biochemistry, 61, 1059–1066.

    Article  CAS  Google Scholar 

  2. Shankar, S. K., Dhananjay, S. K., & Mulimani, V. H. (2009). Applied Biochemistry and Biotechnology, 152, 275–285.

    Article  CAS  Google Scholar 

  3. Weignerová, L., Simerská, P., & Kren, V. (2009). Biocatalysis and Biotransformation, 27, 79–89.

    Article  Google Scholar 

  4. Rodrigues Brasil, A. P., de Rezende, S. T., do Carmo Gouveia Pelúzio, M., & Guimarães, V. M. (2010). Food Chemistry, 118, 251–255.

    Article  Google Scholar 

  5. Machaiah, J. P. P. M. D. (2002). Food Chemistry, 79, 293–301.

    Article  CAS  Google Scholar 

  6. Scalabrini, P., Rossi, M., Spettoli, P., & Matteuzzi, D. (1998). International Journal of Food Microbiology, 39, 213–219.

    Article  CAS  Google Scholar 

  7. Falkoski, D. L., Guimaraes, V. M., Callegari, C. M., Reis, A. P., de Barros, E. G., & de Rezende, S. T. (2006). Journal of Agricultural and Food Chemistry, 54, 10184–10190.

    Article  CAS  Google Scholar 

  8. Kotwal, S. M., Gote, M. M., Khan, M. I., & Khire, J. M. (1999). Journal of Industrial Microbiology & Biotechnology, 23, 661–667.

    Article  CAS  Google Scholar 

  9. Viana, P. A., de Rezende, S. T., Passos, F. M., Oliveira, J. S., Teixeira, K. N., Santos, A. M., et al. (2009). Journal of Agricultural and Food Chemistry, 57, 2515–2522.

    Article  CAS  Google Scholar 

  10. Ganter, C. B. A., Buckel, P., & Mattes, R. (1988). Journal of Biotechnology, 8, 301–310.

    Article  CAS  Google Scholar 

  11. Mulimani, V. H., & Ramalingam. (1995). Biochemistry and Molecular Biology International, 36, 897–905.

    CAS  Google Scholar 

  12. Verdoes, J. C., Punt, P. J., & van den Hondel, C. A. M. J. J. (1995). Applied Microbiology and Biotechnology, 43, 195–205.

    Article  CAS  Google Scholar 

  13. Bergmeyer, H. U., & Bernt, E. (1974). Methods of enzymatic analysis (pp. 1205–1215). New York: Academic Press.

    Google Scholar 

  14. Miller, G. L. (1956). Analytical Chemistry, 31, 426–428.

    Article  Google Scholar 

  15. Bradford, M. M. (1976). Analytical Biochemistry, 72, 248–254.

    Article  CAS  Google Scholar 

  16. Laemmli, U. K. (1970). Nature, 227, 680–685.

    Article  CAS  Google Scholar 

  17. Blum, H., Beier, H., & Gross, H. (1987). Electrophoresis, 8, 93–99.

    Article  CAS  Google Scholar 

  18. McIlvaine, T. C. (1921). The Journal of Biological Biochemistry, 49, 183–186.

    CAS  Google Scholar 

  19. Guimaraes, V. M., de Rezende, S. T., Moreira, M. A., de Barros, E. G., & Felix, C. R. (2001). Phytochemistry, 58, 67–73.

    Article  CAS  Google Scholar 

  20. Ademark, P., de Vries, R. P., Hagglund, P., Stalbrand, H., & Visser, J. (2001). European Journal of Biochemistry, 268, 2982–2990.

    Article  CAS  Google Scholar 

  21. Anisha, G. S., John, R. P., & Prema, P. (2009). Process Biochemistry, 44, 327–333.

    Article  CAS  Google Scholar 

  22. Merja Penttil, A. (1996). European Journal of Biochemistry, 240, 104–111.

    Article  Google Scholar 

  23. Manzanares, P., de Graaff, L. H., & Visser, J. (1998). Enzyme and Microbial Technology, 22, 383–390.

    Article  CAS  Google Scholar 

  24. De Vries, R. P., Van Den Broeck, H. C., Dekkers, E., Manzanares, P., De Graaff, L. H., & Visser, J. (1999). Applied and Environmental Microbiology, 65, 2453–2460.

    Google Scholar 

  25. Puchart, V., Vrsanska, M., Bhat, M. K., & Biely, P. (2001). Biochimica et Biophysica Acta, 1524, 27–37.

    CAS  Google Scholar 

  26. Ramalingam, Saraswathy, N., Sadasivam, S., Subha, K., & Poorani, N. (2007). Indian Journal of Biochemistry & Biophysics, 44, 76–81.

    CAS  Google Scholar 

  27. Viana, P. A., de Rezende, S. T., Marques, V. M., Trevizano, L. M., Passos, F. M., Oliveira, M. G., et al. (2006). Journal of Agricultural and Food Chemistry, 54, 2385–2391.

    Article  CAS  Google Scholar 

  28. Fujimoto, Z., Kaneko, S., Kim, W. D., Park, G. G., Momma, M., & Kobayashi, H. (2009). Bioscience, Biotechnology, and Biochemistry, 73, 2360–2364.

    Article  CAS  Google Scholar 

  29. Mi, S. J., Meng, K., Wang, Y. R., Bai, Y. G., Yuan, T. Z., Luo, H. Y., et al. (2007). Enzyme and Microbial Technology, 40, 1373–1380.

    Article  CAS  Google Scholar 

  30. De Rezende, S. T., Guimarães, V. M., Rodrigues, M. D., & Felix, C. R. (2005). Brazilian Archives of Biology and Technology, 48, 195–202.

    Article  Google Scholar 

  31. Wong, H. C., Hu, C. A., Yeh, H. L., Su, W., Lu, H. C., & Lin, C. F. (1986). Applied and Environmental Microbiology, 52, 1147–1152.

    CAS  Google Scholar 

  32. Oda, Y., & Tonomura, K. (2008). Journal of Applied Microbiology, 80, 203–208.

    Article  Google Scholar 

  33. Cao, Y., Yang, P., Shi, P., Wang, Y., Luo, H., Meng, K., et al. (2007). Enzyme and Microbial Technology, 41, 835–841.

    Article  CAS  Google Scholar 

  34. Prashanth, S. J. M. V. H. (2005). Process Biochemistry, 40, 1199–1205.

    Article  CAS  Google Scholar 

  35. King, M. R., White, B. A., Blaschek, H. P., Chassy, B. M., Mackie, R. I., & Cann, I. K. O. (2002). Journal of Agricultural and Food Chemistry, 50, 5676–5682.

    Article  CAS  Google Scholar 

  36. King, M. R., Yernool, D. A., Eveleigh, D. E., & Chassy, B. M. (1998). FEMS Microbiology Letters, 163, 37–42.

    CAS  Google Scholar 

  37. Nagao, Y., Nakada, T., Imoto, M., Shimamoto, T., Sakai, S., Tsuda, M., et al. (1988). Biochemical and Biophysical Research Communications, 151, 236–241.

    Article  CAS  Google Scholar 

  38. Talbot, G., & Sygusch, J. (1990). Applied and Environmental Microbiology, 56, 3505–3510.

    CAS  Google Scholar 

  39. Dey, P. M., Patel, S., & Brownleader, M. D. (1993). Biotechnology and Applied Biochemistry, 17, 361–371.

    CAS  Google Scholar 

  40. Luonteri, E., Tenkanen, M., & Viikari, L. (1998). Enzyme and Microbial Technology, 22, 192–198.

    Article  CAS  Google Scholar 

  41. Fialho, L. S., Guimaraes, V. M., Callegari, C. M., Reis, A. P., Barbosa, D. S., Borges, E. E., et al. (2008). Phytochemistry, 69, 2579–2585.

    Article  CAS  Google Scholar 

  42. Mahammad, S., Comfort, D. A., Kelly, R. M., & Khan, S. A. (2007). Biomacromolecules, 8, 949–956.

    Article  CAS  Google Scholar 

  43. Civas, A., Eberhard, R., Ledizet, P., & Petek, F. (1984). The Biochemical Journal, 219, 857–863.

    CAS  Google Scholar 

  44. Gote, M., Umlakar, H., Khan, I., & Khire, J. (2004). Process Biochemistry, 39, 1723–1729.

    Article  CAS  Google Scholar 

  45. Rezessy-Szabo, J. M., Nguyen, Q. D., Hoschke, A., Braet, C., Hajos, G., & Claeyssens, M. (2007). Biochimica et Biophysica Acta, 1770, 55–62.

    CAS  Google Scholar 

  46. Civas, A., Eberhard, R., Ledizet, P., & Petek, F. (1984). The Biochemical Journal, 219, 849–855.

    CAS  Google Scholar 

  47. Dey, P. M., & Pridham, J. B. (1972). Advances in Enzymology, 36, 114–118.

    Google Scholar 

  48. Anisha, G. S., Sukumaran, R. K., & Prema, P. (2008). Letters in Applied Microbiology, 46, 338–343.

    Article  CAS  Google Scholar 

  49. De Fátima Viana, S., Guimaraes, V. M., José, I. C., De Almeida E Oliveira, M. G., Brunoro Costa, N. M., De Barros, E. G., et al. (2005). Food Chemistry, 93, 665–670.

    Article  Google Scholar 

  50. Gote, M. M., Khan, M. I., Gokhale, D. V., Bastawde, K. B., & Khire, J. M. (2006). Process Biochemistry, 41, 1311–1317.

    Article  CAS  Google Scholar 

  51. Kachurin, A. M., Golubev, A. M., Geisow, M. M., Veselkina, O. S., Isaeva-Ivanova, L. S., & Neustroev, K. N. (1995). Journal of Biochemistry, 308(Pt 3), 955–964.

    CAS  Google Scholar 

Download references

Acknowledgement

This study was supported by grants of the Fundação de Amparo à Pesquisa do Estado de Minas Gerais-FAPEMIG and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior-CAPES, Brazil.

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Authors and Affiliations

  1. Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, 36.570-000, Viçosa, Minas Gerais, Brazil

    Joana Gasperazzo Ferreira, Angélica Pataro Reis, Valéria Monteze Guimarães, Daniel Luciano Falkoski, Lílian da Silva Fialho & Sebastião Tavares de Rezende

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  1. Joana Gasperazzo Ferreira
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  2. Angélica Pataro Reis
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  3. Valéria Monteze Guimarães
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  5. Lílian da Silva Fialho
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  6. Sebastião Tavares de Rezende
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Correspondence to Sebastião Tavares de Rezende.

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Ferreira, J.G., Reis, A.P., Guimarães, V.M. et al. Purification and Characterization of Aspergillus terreus α-Galactosidases and Their Use for Hydrolysis of Soymilk Oligosaccharides. Appl Biochem Biotechnol 164, 1111–1125 (2011). https://doi.org/10.1007/s12010-011-9198-y

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