Influence of Temperature and pH on Spectral Characteristics of Horseradish Peroxidase in Aqueous Buffer as well as in Nonionic Reverse Micellar System

Anil Kumar

doi:10.14233/ajchem.2015.18863

Issue

Vol. 27 No. 8 (2015): Vol 27 Issue 8

Issue Published : April 27, 2015

This work is licensed under a Creative Commons Attribution 4.0 International License.

Influence of Temperature and pH on Spectral Characteristics of Horseradish Peroxidase in Aqueous Buffer as well as in Nonionic Reverse Micellar System

https://doi.org/10.14233/ajchem.2015.18863

Anil Kumar

Post-Graduate Department of Chemistry, College of Commerce, Patna-800 020, India

Corresponding Author(s) : Anil Kumar

anilcoc2013@gmail.com

Asian Journal of Chemistry, Vol. 27 No. 8 (2015): Vol 27 Issue 8
Article Published : April 28, 2015

Abstract

In this article, temperature dependent spectral characteristics of peroxidase dissolved in aqueous buffer as well as in nonionic reverse micellar systems are reported at different pH values over a wide range of temperature. It is found that the soret band of peroxidase remains unchanged in reverse micellar media as compared to aqueous buffer solution at lower temperature (T £ 20 °C) and then gradually changes as the temperature is increased. At 35 °C the total shape of the spectra changes and at 50 °C it is much more in a distorted form. As the temperature is gradually increased, the total shape of the soret band changes into somewhat random type. This change of the spectral behaviour of horseradish peroxidase at different temperature can be attributed to the structural changes of the enzyme molecule encapsulated inside the aqueous core of the reverse micelles. The absorption spectra of peroxidase with temperature have been studied in reverse micellar media at various W_o. Two types of behaviour of the enzymes are found inside the micellar aqueous core; (i) in the temperature range of 10-30 °C and (ii) in the temperature range 30-50 °C. Again for W_o £ 15 the enzyme behaves in the similar fashion and W > 15 and in the aqueous system, the enzyme behaves in a different fashion. The absorption values slowly increases for W_o £ 15 and attains constant value after that. For W_o = 18 and 20, the absorption value remains constant till the temperature is 30 °C and above that it slowly decreases. The change in absorption spectra of peroxidase molecule with temperature is due to some structural perturbation of the enzyme molecule in reverse micellar media.

Keywords

Horseradish peroxidase Molar extinction coefficient Triton X-100 Reverse micelles Micellar enzymology

Kumar, A. (2015). Influence of Temperature and pH on Spectral Characteristics of Horseradish Peroxidase in Aqueous Buffer as well as in Nonionic Reverse Micellar System. Asian Journal of Chemistry, 27(8), 3057–3062. https://doi.org/10.14233/ajchem.2015.18863

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References

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References

(i) S. Roy, A. Dasgupta and P.K. Das, Langmuir, 22, 4567 (2006); doi:10.1021/la0602867; Biswas, A.R. Das, T. Pradhan, D. Touraud, W. Kunz and S. Mahiuddin, J. Phys. Chem. B, 112, 6620 (2008); doi:10.1021/jp711368p.

K. Holmberg, Adv. Colloid Interface Sci., 51, 137 (1994); doi:10.1016/0001-8686(94)80035-9.

M. Moniruzzaman, N. Kamiya and M. Goto, Langmuir, 25, 977 (2009); doi:10.1021/la803118q.

A.S. Bommarius, T.A. Hatton and D.I.C. Wang, J. Am. Chem. Soc., 117, 4515 (1995); doi:10.1021/ja00121a010.

F. Franks, Water A Comprehensive Treatise, Plenum, New York (1975).

C.M.L.Carvalho and J.M.S.Cabral, Biochimie, 82, 1063 (2000); doi:10.1016/S0300-9084(00)01187-1.

P.L.Luisi, L.Magid and J.H.Fendler, CRC Crit. Rev. Biochem., 20, 409 (1986); doi:10.3109/10409238609081999.

(i) K. Martinek, A.V. Levashov, Y.L. Khmelnitsky, N.L. Klyachko and I.V. Berezin, Science, 218, 889 (1982); doi:10.1126/science.6753152; (ii) N.L. Klyachko, A.V. Levashov and K. Martinek, J. Mol. Biol., 18, 830 (1984).

R.P. Cullis, B. Kruijff, M.J. Hope, R. Nayar and S.L. Schmid, Can. J. Biochem., 58, 1091 (1980); doi:10.1139/o80-147.

R.M.C. Dawson, J. Am. Oil Chem. Soc., 59, 401 (1982); doi:10.1007/BF02634422.

S. Sarcar, T.K. Jain and A. Maitra, Biotechnol. Bioeng., 39, 474 (1992); doi:10.1002/bit.260390416.

A.C. Maehly, Biochem. Biophys. Acta, 8, 1 (1952); doi:10.1016/0006-3002(52)90002-4.

(i) K. Martinek, A.V. Levashov and N.L. Klyachko, Molecular Biology (Russ), 18, 1019 (1984); (ii) Yu. L. Khel’nitskii, A.V. Levashov, N.L. Klyachko, V.Ya. Chernyak and K. Martinek, Biockimiya, 47, 86 (1981).

A. Kumar, Ph.D. Thesis, University of Delhi, India (1999).

D. Kelin and T. Mann, Proc. Roy. Soc., 22, 118 (1937).

H. Theorell, Arkiv. Kemi. Mineral Geol., 16A, (1942).

F.M. Menger and G. Saito, J. Am. Chem. Soc., 100, 4376 (1978); doi:10.1021/ja00482a010.

A.V. Levashov, N.L. Klyachko and K. Martinek, Bioorg. Kemi (Strasbg.), 7, 670 (1981).

A.C. Maehly, Methods Enzymol., 2, 801 (1955); doi:10.1016/S0076-6879(55)02307-0.

S. Das and A. Maitra, Colloids Surf., 35, 101 (1989); doi:10.1016/0166-6622(89)80323-3.

D.M. Zhu, X. Wu and Z.A. Schelly, Langmuir, 8, 1538 (1992); doi:10.1021/la00042a008.

H. Theorell, A.C. Maehly, H. Dam and P.-O. Kinell, Acta Chem. Scand., 4, 422 (1950); doi:10.3891/acta.chem.scand.04-0422.

Issue

Vol. 27 No. 8 (2015): Vol 27 Issue 8

Influence of Temperature and pH on Spectral Characteristics of Horseradish Peroxidase in Aqueous Buffer as well as in Nonionic Reverse Micellar System

Corresponding Author(s) : Anil Kumar

Abstract

Keywords

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