Abstract
Thermal stability of horseradish peroxidase (HRP) was studied by differential scanning calorimetry, tryptophan fluorescence, the heme absorption and enzymatic activity analysis while the concentrations of sodium phosphate buffer ranged from 2.5 to 50 mM at pH 7.0.
The results showed that the denaturation temperature (T m) values decreased and the intrinsic tryptophan fluorescence intensity of denatured HRP increased as sodium phosphate buffer concentration increased. Furthermore, the heme absorbance at 403 nm and enzymatic activity of HRP decreased with the increasing buffer concentrations. According to data obtained in this experiment, it can be concluded that sodium phosphate accelerated the denaturation process of HRP and reduced the thermal stability of HRP.
Similar content being viewed by others
References
H. B. Dunford, Ed., Heme Peroxidases, Wiley, New York 1999.
P. Tijssen and E. Kurstak, Anal. Biochem., 136 (1984) 451.
R. Maidan and A. Heller, J. Am. Chem. Soc., 113 (1991) 9003.
M. G. Garguilo, N. Huynh, A. Proctor and C. A. Michael, Anal. Chem., 65 (1993) 523.
A. L. Ghindilis, P. Atanasov and E. Wilkins, Electroanalysis, 9 (1997) 661.
M. Gajhede, D. J. Schuller, A. Henriksen, A. T. Smith and T. L. Poulos, Nat. Struct. Biol., 4 (1997) 1032.
O. Mirza, A. Henriksen, L. Ostergaard, K. G. Welinder and M. Gajhede, Acta Crystallogr. D: Biol. Crystallogr., 56 (2000) 372.
A. S. L. Carvalho, E. P. E. Melo, B. S. Ferreira, M. T. Neves-Petersen, S. B. Petersen and M. R. Aires-Barros, Arch. Biochem. Biophys., 415 (2003) 257.
H. S. Pappa and A. E. G. Cass, Eur. J. Biochem., 212 (1993) 227.
K. G. Welinder, J. M. Mauro and L. Norskov-Lauritsen, Biochem. Soc. Trans., 20 (1992) 337.
T. L. Poulos, Curr. Opin. Biotechnol., 4 (1993) 484.
A. M. English and G. Tsaprailis, Adv. Inorg. Chem., 43 (1995) 79.
A. Henriksen, O. Mirza, C. Indiani, K. Teilum, G. Smulevich, K. G. Welinder and M. Gajhede, Protein Sci., 10 (2001) 108.
N. C. Veitch and R. J. P. Williams, Eur. J. Biochem., 189 (1990) 351.
J. W. Tams and K. G. Welinder, Biochemistry, 35 (1996) 7573.
G. Tsaprailis, D. W. S. Chan and A. M. English, Biochemistry, 37 (1998) 2004.
K. Chattopadhyay and S. Mazumdar, Biochemistry, 39 (2000) 263.
D. G. Pina, A. V. Shnyrova, F. Gavilanes, A. Rodriguez, F. Leal, M. G. Roig, I. Y. Sakharov, G. G. Zhadan, E. Villar and V. L. Shnyrov, Eur. J. Biochem., 268 (2001) 120.
A. S. L. Carvalho, M. T. Neves-Petersen, S. B. Petersen, M. R. Aires-Barros and E. P. E. Melo, Biochim. Biophys. Acta, 1747 (2005) 99.
A. S. L. Carvalho, B. S. Ferreira, M. T. Neves-Petersen, S. B. Petersen, M. R. Aires-Barros and E. P. Melo, Enzyme Microb. Technol., 2006 in press.
G. Rialdi and E. Battistel, J. Thermal Anal., 47 (1996) 17.
A. Ginsburg, J. Therm. Anal. Cal., 61 (2000) 425.
G. D. Manetto, C. La Rosa, D. M. Grasso and D. Milardi, J. Therm. Anal. Cal., 80 (2005) 263.
A. Michnik, K. Michalik and Z. Drzazga, J. Therm. Anal. Cal., 80 (2005) 399.
B. Bugyi, G. Papp, S. Halasi and B. Visegrády, J. Therm. Anal. Cal., 82 (2005) 275.
G. Papp, B. Bugyi, Z. Ujfalusi, S. Halasi and J. Orbán, J. Therm. Anal. Cal., 82 (2005) 281.
A. Michnik, J. Therm. Anal. Cal., 87 (2007) 91.
A. Stirpe, R. Guzzi, H. Wijma, M. P. Verbeet, G. W. Canters and L. Sportelli, Biochim. Biophys. Acta, 1752 (2005) 47.
M. Rezaei-Tavirani, S. H. Moghaddamnia, B. Ranjbar, M. Amani and S. A. Marashi, J. Biochem. Mol. Biol., 39 (2006) 530.
X. P. Geng, Y. N. Wu, B. H. Wang, H. F. Zhang, X. D. Geng and J. W. Xing, J. Therm. Anal. Cal., 85 (2006) 601.
J. W. Tams and K. G. Welinder, FEBS Lett., 421 (1998) 234.
M. A. Lemos, J. C. Oliveira and J. A. Saraiva, Lebensmittel-Wissenschaft und-Technologie, 33 (2000) 362.
R. Lumry and E. Eyring, J. Phys. Chem., 58 (1954) 110.
J. M. Sanchez-Ruiz, Biophys. J., 61 (1992) 921.
I. M. Plaza del Pino, B. Ibarra-Molero and J. M. Sanchez-Ruiz, Proteins, 40 (2000) 58.
H. F. Liu, Z.Y. Wang, Y. W. Liu, J. Xiao and C. X. Wang, Thermochim. Acta, 443 (2006) 173.
J. E. Brunet, G. A. Gonzalez and C. P. Sotomayor, Photochem. Photobiol., 38 (1983) 253.
T. K. Das and S. Mazumdar, Eur. J. Biochem., 227 (1995) 823.
J. R. Lakowicz, Principles of Fluorescence Spectroscopy, Plenum Press, New York 1983.
A. T. Smith and N. C. Veitch, Curr. Opin. Chem. Biol., 2 (1998) 269.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Haifeng, L., Yuwen, L., Xiaomin, C. et al. Effects of sodium phosphate buffer on horseradish peroxidase thermal stability. J Therm Anal Calorim 93, 569–574 (2008). https://doi.org/10.1007/s10973-007-8407-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10973-007-8407-y