Supermacroporous Support Having Dye for Reuseable Enzyme Immobilization

Year 2008, Volume: 36 Issue: 1, 9 - 19, 01.01.2008

M. Nalan Tüzmen Nilay Bereli Müge Andaç Sinan Akgöl Adil Denizli

Abstract

Poly acrylamide-allyl glycidyl ether [poly AAm-AGE ] cryogel was prepared by bulk polymerization which proceedsin an aqueous solution of monomers frozen inside a glass column cryo-polymerization . After thawing, the monolithiccryogel contains a continuous polymeric matrix having interconnected pores of 10-100 µm size. Cibacron Blue F3GAwas attached by covalent binding onto poly AAm-AGE cryogel via epoxy groups. Poly AAm-AGE cryogel wascharacterized by FTIR and scanning electron microscopy SEM . The equilibrium swelling degree of the poly AAmAGE monolithic cryogel was 6.84 g H2O/g cryogel. Poly AAm-AGE cryogel containing 68.9 µmol Cibacron BlueF3GA/g was used in the adsorption ofβ-casein from aqueous solution. The effects of initial concentration, pH, ionicstrength and flow rate on the adsorption efficiency of cryogel were studied in a column system. The non-specificadsorption ofβ-casein was very low 1.26 mg/g polymer . The maximum amount of β-casein adsorption from aqueoussolution in acetate buffer was 44.7 mg/g at pH 4.0. Up to 99% of the adsorbedβ-casein was eluted when 0.1 MTris/HCl buffer containing 0.5 M NaCl was used as elution agent. We concluded that Cibacron Blue F3GA attachedpoly AAm-AGE cryogel could be repeatedly applied forβ-casein adsorption without significant losses in the adsorptioncapacity

Keywords

β-Casein, cryogels, Dye-affinity Chromatography, Protein Purification

References

• 1. G. M. S. Finette, Q. M. Mao, M. T. W. Hearn, J. Chromatogr. A., 763 (1997) 71.
• 2. S. Fisichella, G. Alberghina, M. E. Amato, D. Lafiandra, D.Mantarro, A. Palermo, Savarino, G.Scarlata, J. Cereal Sci., 38 (2003) 77.
• 3. M.Grasselli, S. A. Camperi, d A. A. N.el Canizo, O.Cascone, J.Sci. Food and Agricul., 79 (1999) 333.
• 4. S. Zhang, Y. Sun, Biotechnol. Prog., 20 (2004) 207.
• 5. E. P. Lillehoj, V. S. Malik, in: Advances in Biochemical Engineering/Biotechnology, A. Fiechter, Ed., Springer-Verlag, Berlin 1982, pp. 20– 65.
• 6. Y. Arica, H. N. Testereci, A. Denizli, J. Chromatogr. A , 799 (1998) 83.
• 7. S. Akgol, D. Turkmen, A. Denizli, J. Appl. Polym. Sci., 93 (2004) 2669.
• 8. S. Senel, S. Akgol,Y. Arica, A. Denizli, Polym. Int., 50 (2001) 1143.
• 9. S. Akgol, A. Denizli, J. Mol. Cat. B: Enzym., 28 (2004) 7.
• 10. M. Odabasi, A. Denizli, Polym. Int., 53 (2004) 332.
• 11. H. Yavuz, A. Denizli, Macromol. Biosci., 5 (2005) 39.
• 12. H. Yavuz, A. Denizli, Macromol. Biosci., 4 (2004) 84.
• 13. R.M. Birch, C. O’Byrne, I.R. Booth, P. Cash, Proteomics, 3 (2003) 764.
• 14. A. Denizli, E. Pişkin, J. Biochem. Biophys. Methods, 49 (2001) 391.
• 15. M. Odabasi, A. Denizli, Polym. Int., 53 (2004) 332.
• 16. S. Zhang, Y. Sun, Ind. Eng. Chem. Res., 42 (2003) 1235.
• 17. M. Zhang, Y. Sun, Biotechnol. Bioeng., 75 (2001) 710.
• 18. M. Zhang, Y. Sun, J. Chromatogr. A., 912 (2001) 31.
• 19. H. Yavuz, S. Akgöl, R. Say, A. Denizli, Int.J. Biol. Macromol., 39 (2006) 303.
• 20. S. Akgöl, A. Denizli, J. Molecular Catalyis B., 28 (2004) 7.
• 21. M. McCoy, K. Kalghatgi, F.E. Regnier, N. Afeyan, J. Chromatogr. A., 743 (1996) 221.
• 22. E.B. Altıntas, A. Denizli, J. Appl. Polym. Sci., 103 (2007) 975.
• 23. S. Özkara, B. Garipcan, E. Piflkin, A. Denizli, J. Biomater. Sci. Polym. Ed., 14 (2003) 761.
• 24. V.I. Lozinsky, I.Y. Galaev, F.M. Plieva, I.N. Savina, H. Jungvid, B. Mattiason, Trends in Biotechnol., 21 (2003) 445.
• 25. P. Arvidsson, F.M. Plieva, V.I. Lozinsky, I.Y. Galaev, B. Mattiasson, J. Chromatogr. A., 986 (2003) 275.
• 26. V.I. Lozinsky, F.M. Plieva, I. Y. Galaev, B. Mattiasson, Bioseparation, 10 (2002) 163.
• 27. P. Arvidsson, F.M. Plieva, I.N. Savina, V.I. Lozinsky, S. Fexby, L. Bülow, I.Y. Galaev, B. Mattiasson, J. Chromatogr. A., 977 (2002) 27.
• 28. C. Babaç, H. Yavuz, I.Y. Galaev, E. Piflkin, A. Denizli, React. Functl. Polym., 66 (2006) 1263.
• 29. M.B. Dainiak, I.Y. Galaev, B. Matiasson, J. Chromatogr. A., 1123 (2006) 145.
• 30. I.M. Savina, F.Plieva, V.A. Izumrudov, B. Mattiasson, I. Y. Galaev, J. Biotechnol., 123 (2006) 209.
• 31. N. Demiryas, N. Tüzmen, I.Y. Galaev, E. Pişkin, A. Denizli, J. Appl. Polym. Sci., in press.
• 32. C. G. De Kruif, V. Y. Grinberg, Coll. Surf. A., 210 (2002) 183.
• 33. K. Landström, T. Arnebrant, J. Alsins, B. Bergenstahl, Food Hydrocolloids, 17 (2003), 103- 116.
• 34. C. G. De Kruif, in: Industrial Proteins,W. Aalbersberg, Ed., Elsevier Sci., London 2003, Chapter 6.
• 35. E. Dickinson, S. Krishna, Food Hydrocolloids, 15 (2001) 107-115.
• 36. M. R. R. Nino, C. C. Sanchez, J. M. R. Patino, Coll. Surf. B., 12 (1999) 161.
• 37. H. E. Swaisgood, Adv. Dairy Chem., 1 (1992) 63.
• 38. S. Akgöl, N. Bereli, A. Denizli, Macromol. Biosci. 5 (2005), 786-794.
• 39. Y. Igarashi, Int. Dairy J., 9 (1999) 377.
• 40. P. Arvidsson, F.M. Plieva, I.N. Savina, V.I. Lozinsky, S. Fexby, L. Bülow, I.Y. Galaev, B. Mattiasson, J. Chromatogr. A., 977 (2002) 27.
• 41. B. Weber, K. Willeford, J. Moe, D. Piskiewicz, Biochem. Biophys. Res. Commun., 86 (1979) 252.
• 42. C.V. Stead, in: Y.D. Clonis, A. Atkinson, C.J. Bruton, C.R. Lowe (Eds.), Reactive Dyes in Protein and Enzyme Technology, Macmillan, Basingstoke, 1987, p. 13.
• 43. P.M. Boyer, J.T. Hsu, Adv. Biochem. Eng., 49 (1993) 1.
• 44. C.R. Lowe, in: A. Wiseman (Ed.), Topics in Enzyme and Fermentation Technology, Ellis Horwood, Chichester 1984, p. 78.
• 45. X. Zhu, S.D. Alexandratos, Ind. Eng. Chem. Res., 44 (2005) 8605.
• 46. Y.S. Ho, G. McKay, Proc. Biochem., 34 (1999) 451.
• 47. L. Uzun, A. Denizli, J. Biomater. Sci. Polym. Ed., 17 (2006) 791.
• 48. S.J. Allen, B. Koumanova, Z. Kircheva, S. Nenkova, Ind. Eng. Chem. Res., 44 (2005) 2281.
• 49. S. Oncel, L. Uzun, B. Garipcan, A. Denizli, Ind. Eng. Chem. Res., 44 (2005) 7049.