Unique Activity of Ureases Immobilized on Poly(Styrene-Co-Acrolein) Microspheres

Abstract

Poly(styrene-co-acrolein) (P(S/A)) microspheres were synthesized by emulsifier-free emulsion-dispersion radical copolymerization of styrene and acrolein. Particles with a number average diameter (øD _n) of 373 nm, a narrow diameter distribution (øD _w/øD _n) of 1.008 and a surface concentration of aldehyde groups from polyacrolein units of 2.51 · 10⁻⁶ mol/m² were obtained. Ureases from jack beans (Urs-JB) and from pigeonpea (Urs-PP) were immobilized onto these microspheres. Activity of free and immobilized urease was determined using a standard phenol method. Surface concentration of attached enzymes was varied in the range from 0.06 to 1.6 mg/m². Specific activity of Urs-JB in solution was 1.9 times higher than that of Urs-PP, however, upon immobilization of enzymes onto P(S/A) microspheres the reverse was true. Due to very high denaturation activity of immobilized Urs-JB was 10 times lower than in solution. In the case of Urs-PP, denaturation of the enzyme upon immobilization was much lower and possibly due to a cooperative effect resulting from a local high concentration of immobilized enzyme, its specific activity was from 3 to 9 times higher than that of immobilized Urs-JB.

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References

1. Roig, M.G., Bello, J.F., Rodriguez, S., Kennedy, J.F. and Taylor, D.W. (1966). Polym. Int., 39: 17 .

2. Guilbault, G.G. (1976). Handbook of Enzymatic Methods of Analysis, pp. 1-738, Marcel Dekker, New York, Basel .

3. Edelman, P.G. and Wang, J. (eds), (1992). Biosensors and Chemical Sensors, Optimizing Performance Through Polymeric Materials, ACS Symp. Ser., Vol. 487, pp. 1-332, Washington, DC .

4. Wu, C.-W., Lee, J.-G., and Lee, W.-C. (1988). Biotechnol. Appl. Biochem., 27: 225 .

5. Barker, S.A. (1987). The Diagnosis of Human Genetic Disease in Biosensors, In: Turner, A.P.F., Karube, I. and Wilson, G.S. (eds), Fundamentals and Applications, p. 71, Oxford University Press, New York .

6. Colvin, M., Smolka, A., Rembaum, A. and Chang, M. (1988). The Covalent Binding of Enzymes and Immunoglobulins to Hydrophilic Microspheres in Microspheres: Medical and Biological Applications, In: Rembaum, A. and Tokes, Z.A. (eds), Chapter 1, p. 1, CRC Press, Inc., Boca Raton, Florida .

7. Okuma, H., Sekimukai, S., Hoshi, M., Toyama, K. and Watanabe, E. (1989). Enzyme Microb. Technol., 11: 824 .

8. Nicolae, C.A., Cantin-Riviere, S., El Abed, A. and Peretti, A.P. (1997). Langmuir, 13: 5507 .

9. Das, N., Prabhakar, P., Kayastha, A.M. and Srivastava, R.C. (1997). Biotechnol. Bioeng., 54: 329 .

10.

10. Lukachova, L.V., Karyakin, A.A., Karyakina, E.E. and Gorton, L. (1997). Sensors Actuators B, 44: 356 .

11.

11. Santucci, M., Portaccio, M., Rossi, S., Bencivenga, U., Gaeta, F.S. and Mitab, D.G. (1999). Biosens. Bioelectron., 14: 737 .

12.

12. Yao, T. and Takashima, K. (1998). Biosens. Bioelectron., 13: 67 .

13.

13. Adeloju, S.B. and Moline, A.N. (2001). Biosens. Bioelectron., 16: 133 .

14.

14. Brahim, S., Narinesingh, D. and Guiseppi-Elie, A. (2001). Biosens. Bioelectron., 17: 53 .

15.

15. Andrade, J.D. and Hlady, V. (1986). Adv. Polym. Sci., 79: 1 .

16.

16. Norde, W., MacRitchie, F., Nowicka, G. and Lyklema, J. (1986). Colloid Interface Sci., 112: 447 .

17.

17. Basinska, T., Kowalczyk, D., Miksa, B. and Slomkowski, S. (1995). Polymers Adv. Technol., 6: 526 .

18.

18. Maste, M.C.L., Norde, W. and Visser, A.J.W.G. (1997). J. Colloid Interface Sci., 196: 224 .

19.

19. Buijs, J. and Hlady, V. (1997). J. Colloid Interface Sci., 190: 171 .

20.

20. Pye, E.K. and Chance, B. (1976). Investigation of the Physical Properties of Immobilized Enzymes in Immobilized Enzymes, In: Mosbach, K. (ed.), Methods in Enzymology, Vol. 44, p. 357, Academic Press, New York .

21.

21. Chang, M., Colvin, M. and Rembaum, A. (1986). J. Polym. Sci. Polym. Letters., Ed., 24, 603 .

22.

22. Chen, J.P., Kiaei, D. and Hoffman, A.S. (1993). J. Biomater. Sci. Polymer Edn., 5: 167 .

23.

23. Basinska, T. and Slomkowski, S. (1995). Colloid and Polymer Sci., 273: 431 .

24.

24. Basinska, T., Slomkowski, S. and Delamar, M. (1993). J. Bioact. Compat. Polym., 8: 205 .

25.

25. Basinska, T. and Slomkowski, S. (1993). Polystyrene and Poly(styrene/acrolein) Latexes with Immobilized Proteins as a Basis of Diagnostic Agglutination Test in Uses of Immobilized Biological Compounds, In: Guilbault, G. and Mascini, M., (eds), p. 453, Kluwer Academic Publishers, Dordrecht, Boston, London .

26.

26. Slomkowski, S. (1998). Progr. Polym. Sci., 23: 815 .

27.

27. Das, N., Kayastha, A.M. and Malhotra, O.P. (1998). Biotechnol. Appl. Biochem., 27: 25 .

28.

28. Das, N. and Kayastha, A.M. (1998). World J. Microbiol. Biotechnol., 14: 927 .

29.

29. Srivastava, P.K., Kayastha, A.M. and Srinivasan (2001). Biotechnol. Appl. Biochem., 34: 55 .

30.

30. Kayastha, A.M. and Srivastava, P.K. (2001). Appl. Biochem. Biotechnol., 96: 41 .

31.

31. Basinska, T. and Slomkowski, S. (1991). J. Biomater. Sci. Polymer Edn., 3: 115 .

32.

32. Slomkowski, S. and Basinska, T. (1992). Detection and Concentration Measurements of Proteins Immobilized on the Polystyrene and Poly (Styrene/Acrolein) Latexes in Polymer Latexes, Preparation, Characterization, and Applications , In: Daniels, E.S., Sudol, E.D. and El-Aasser, M.S. (eds), ACS Symp. Ser., 492: 328 .

33.

33. Das, N., Kayastha, A.M. and Srivastava, P.K. (2002). Phytochemistry, 61: 513 .

34.

34. Fawcett, J.K. and Scott, J.E. (1960). J. Clin. Path., 13: 1565 .

35.

35. Miksa, B. and Slomkowski, S. (1995). J. Biomater. Sci. Polym. Edn., 7: 77 .

36.

36. Slomkowski, S., Kowalczyk, D., Chehimi, M.M. and Delamar, M. (2000). Colloid Polym. Sci., 278: 878 .

37.

37. Fishbein, W. and Daly, J. (1970). Proc. Soc. Exp. Biol., 134: 1083 .

38.

38. Fishbein, W. and Nagarajan, K. (1972). Arch. Biochem. Biophys., 151: 370 .

39.

39. Kowalczyk, D., Marsault, J.-P. and Slomkowski, S. (1996). Colloid Polym. Sci., 274: 513 .