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Immobilization of Aspergillus oryzae β-galactosidase on low-pressure plasma-modified cellulose acetate membrane using polyethyleneimine for production of galactooligosaccharide

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Abstract

The aim of this study was to produce galactooligosaccharides (GOS) from lactose using β-galactosidase from Aspergillus oryzae immobilized on a low-pressure plasma-modified cellulose acetate (CA) membrane. Specifically, a novel method was developed for multilayer enzyme immobilization involving polyethyleneimine (PEI)-enzyme aggregate formation and growth on a CA membrane. A large amount of enzyme (997 μg/cm2 membrane) was immobilized with 66% efficiency. The K m value for the immobilized enzyme was estimated to be 48 mM, which indicates decreased affinity for the substrate, whereas the Vmax value was smaller. The immobilized enzyme showed good storage and operational stability. The half-life of the immobilized enzyme on the membrane was about 1 month at 30°C and ∼ 60 h at 60°C. Maximum GOS production of 27% (w/w) was achieved with 70% lactose conversion from 320 g/L of lactose at pH 4.5 and 60°C. Trisaccharides were the major types of GOS formed and accounted for about 75% of the total GOS produced. Based on these results, immobilized enzyme technology could be applied to GOS production from lactose.

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References

  1. Sako, T., K. Matsumoto, and R. Tanaka (1999) Recent progress on research and applications of non-digestible galacto-oligosac-charides. Intl. Dairy J. 9: 69–80.

    Article  CAS  Google Scholar 

  2. Gaur, R., H. Pant, R. Jain, and S. K. Khare (2006) Galacto-oligosaccharide synthesis by immobilized Aspergillus oryzae β-galactosidase. Food Chem. 97: 426–430.

    Article  CAS  Google Scholar 

  3. Albayrak, N. and S. T. Yang (2002) Immobilization of β-galactosidase on fibrous matrix by polyethylenimine for production of galacto-oligosaccharides from lactose. Biotechnol. Prog. 18: 240–251.

    Article  CAS  Google Scholar 

  4. Haider, T. and Q. Husain (2009) Immobilization of β-galactosidase from Aspergillus oryzae via immunoaffinity support. Biochem. Eng. J. 43: 307–314.

    Article  CAS  Google Scholar 

  5. Shin, H. J., J. M. Park, and J. W. Yang (1998) Continuous production of galactooligosaccharides from lactose by Bullera singularis β-galactosidase immobilized in chitosan bead. Proc. Biochem. 33: 787–792.

    Article  CAS  Google Scholar 

  6. Kim, H. I. and S. S. Kim (2001) Fabrication of reverse osmosis membrane via low temperature plasma polymerization. J. Memb. Sci. 190: 21–33.

    Article  CAS  Google Scholar 

  7. Güleç, H. A., K. Saroğlu, and M. Mutlu (2006) Modification of food contacting surfaces by plasma polymerisation technique. Part I: Determination of hydrophilicity, hydrophobicity and surface free energy by contact angle method. J. Food Eng. 75: 187–195.

    Article  Google Scholar 

  8. Gulec, H. A., A. Topacli, C. Topacli, N. Albayrak, and M. Mutlu (2010) Modification of cellulose acetate membrane via low-pressure plasma polymerization for sugar separation applications: Part I. Membrane development and characterization. J. Memb. Sci. 350: 310–321.

    Article  CAS  Google Scholar 

  9. Foda, M. I. and M. Lopez-Leiva (2000) Continous production of oligosaccharides from whey using a membrane reactor. Proc. Biochem. 35: 581–587.

    Article  CAS  Google Scholar 

  10. Czermak, P., M. Ebrahimi, K. Grau, S. Netz, G. Sawatzki, and P. H. Pfromm (2004) Membrane-assisted enzymatic production of galactosyl-oligosaccharides from lactose in a continuous process. J. Memb. Sci. 232: 85–91.

    Article  CAS  Google Scholar 

  11. Matuschewski, H. and U. Schedler (2008) MSE-modified membranes in organophilic pervaporation for aromatics/aliphatics separation. Desalination 224: 124–131.

    Article  CAS  Google Scholar 

  12. Zemek, J., L. Kuniak, P. Gemeiner, J. Zamocky, and S. Kucar (1982) Cross-linked polyethylenimine: an enzyme carrier with spacers of various lengths introduced in cross-linking reaction. Enz. Microb. Technol. 4: 233–238.

    Article  CAS  Google Scholar 

  13. Jendrisak, J. (1987) The use of polyethyleneimine in protein purification. UCLA Symp. Mol. Cell. Biol. New Ser. 68: 75–97.

    CAS  Google Scholar 

  14. Wasserman, B. P. and H. O. Hultin (1980) High-yield method for immobilization of enzymes. Biotechnol. Bioeng. 22: 271–287.

    Article  CAS  Google Scholar 

  15. Emneus, J. and L. Gorton (1993) Comparison between different inorganic supports for the immobilization of amyloglucosidase and α-amylase to be used in enzyme reactors in flow-injection systems: Part II. Hydrolysis of glycogen. Anal. Chim. Acta 276: 319–328.

    Article  CAS  Google Scholar 

  16. Yang, S. T., J. L. Marchio, and J. W. Yen (1994) A Dynamic light scattering study of β-galactosidase: Environmental effects on protein conformation and enzyme activity. Biotechnol. Prog. 10: 525–531.

    Article  CAS  Google Scholar 

  17. Reshmi, R., G. Sanjay, and S. Sugunan (2007) Immobilization of α-amylase on zirconia: A heterogeneous biocatalyst for starch catalysis. Catal. Commun. 8: 393–399.

    Article  CAS  Google Scholar 

  18. Dissing, U. and B. Mattiasson (1996) Polyelectrolyte complexes as vehicles for affinity precipitation of proteins. J. Biotechnol. 52: 1–10.

    Article  CAS  Google Scholar 

  19. Onda, M., K. Ariga, and T. Kunitake (1999) Activity and stability of glucose oxidase in molecular films assembled alternately with polyions. J. Biosci.Bioeng. 87: 69–75.

    Article  CAS  Google Scholar 

  20. Tanriseven, A. and S. Dogan (2002) A novel method for immobilization of β-galactosidase. Proc. Biochem. 38: 27–30.

    Article  CAS  Google Scholar 

  21. Zhou, Q. Z. K. and X. D. J. Chen (2001) Immobilization of β-galactosidase on graphite surface by gluteraldehyde. J. Food Eng. 48: 69–74.

    Article  Google Scholar 

  22. Bayramoglu, G., Y. Tunali, and M. Y. Arica (2007) Immobilization of β-galactosidase onto magnetic poly(GMA-MMA) beads for hydrolysis of lactose in bed reactor. Catal. Commun. 8: 1094–1101.

    Article  CAS  Google Scholar 

  23. Taqieddin, E. and M. Amiji (2004) Enzyme immobilization in novel alginate-chitosan core-shell microcapsules. Biomaterials 25: 1937–1945.

    Article  CAS  Google Scholar 

  24. El-Masry, M. M., A. De Maio, P. L. Martelli, R. Casadio, A. B. Moustafa, S. Rossi, and D. G. Mita (2001) Influence of the immobilization process on the activity of β-galactosidase bound to Nylon membranes grafted with glycidyl methacrylate: Part 1. Isothermal behavior. J. Mol. Catal. B: Enz. 16: 175–189.

    Article  CAS  Google Scholar 

  25. Berger, J. L., B. H. Lee, and C. Lacroix (1995) Oligosaccharides synthesis by free and immobilized β-galactosidases from Thermus aquaticus YT-1. Biotechnol. Lett. 17: 1077–1080.

    Article  CAS  Google Scholar 

  26. Mozaffar, Z., K. Nakanishi, R. Matsuno, and T. Kamikuba (1984) Production and properties of β-galactosidases from Bacillus circulans. Agric. Biol. Chem. 48: 3053–3061.

    CAS  Google Scholar 

  27. Mozaffar, Z., K. Nakanishi, and R. Matsuno (1986) Continuous production of galacto-oligosaccharides from lactose using immobilized β-galactosidase from Bacillus circulans. Appl. Microbiol. Biotechnol. 25: 224–228.

    CAS  Google Scholar 

  28. Iwasaki, K., M. Nakajima, and S. Nakao (1996) Galacto-oligosaccharide production from lactose by an enzymatic batch reaction using β-galactosidase. Proc. Biochem. 31: 69–76.

    Article  CAS  Google Scholar 

  29. Albayrak, N. and S. T. Yang (2002) Production of galacto-oligosaccharides from lactose by Aspergillus oryzae β-galacosidase immobilized on cotton cloth. Biotechnol. Bioeng. 77: 8–19.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Food Engineering, Faculty of Engineering and Architecture, Trakya University, Edirne, 22180, Turkey

    Hacı Ali Güleç

  2. Plasma Aided Bioengineering and Biotechnology Research Group, Hacettepe University, Ankara, 06800, Turkey

    Hacı Ali Güleç, Sevim Gürdaş & Mehmet Mutlu

  3. Department of Food Engineering, Faculty of Engineering, Cumhuriyet University, Sivas, 58140, Turkey

    Sevim Gürdaş

  4. Department of Food Engineering, Faculty of Engineering, Hitit University, Çorum, 19030, Turkey

    Nedim Albayrak

Authors
  1. Hacı Ali Güleç
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  2. Sevim Gürdaş
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  3. Nedim Albayrak
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  4. Mehmet Mutlu
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Correspondence to Hacı Ali Güleç.

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Güleç, H.A., Gürdaş, S., Albayrak, N. et al. Immobilization of Aspergillus oryzae β-galactosidase on low-pressure plasma-modified cellulose acetate membrane using polyethyleneimine for production of galactooligosaccharide. Biotechnol Bioproc E 15, 1006–1015 (2010). https://doi.org/10.1007/s12257-010-0046-7

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  • DOI: https://doi.org/10.1007/s12257-010-0046-7

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