Abstract
Hemoglobin, horseradish peroxidase, and bovine serum albumin incubated heme-catalyzed the oxidation of dibenzothiophene into sulfoxide in the presence of hydrogen peroxide. This reaction was carried out in an aqueous buffer containing 25% of water-miscible organic solvents. The observation of this transient state of hemoproteins during sulfoxidation showed heme degradation. None of the compounds usually involved in a classical peroxidative activity mechanism were detected. Furthermore, this activity did not appear to be based on a Fenton-type reaction. The highest degrees of sulfoxidation were obtained with hemoglobin. Under the best conditions of reaction, 100% of dibenzothiophene were converted into dibenzothiophene sulfoxide by hemoglobin. Heat-denatured hemoproteins did keep their sulfoxidation activity. With hemoglobin, a kcat of 0.22 min-1 was determined. Nearly the same values were obtained with heat-denatured hemoglobin and bovine serum albumin-adsorbed heme. With horseradish peroxidase, only 4% of conversion was attained. This percentage could be slightly increased by using a less pure peroxidase or heat-denatured peroxidase.
Similar content being viewed by others
Abbreviations
- Heme:
-
iron protoporphyrin IX regardless of the oxidation and ligation state
- Hb:
-
hemoglobin
- HRP:
-
horseradish peroxidase
- DBT:
-
dibenzothiophene
- SOD:
-
Superoxide dismutase
- BSA:
-
bovine serum albumin
- ABTS:
-
2,2−azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid)
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- DMF:
-
N,N-dimethylformamide
- Hemin:
-
chloroprotoporphyrin IX iron (III)
- HPLC:
-
high-pressure liquid chromatography
- GC/MS:
-
gas chromatography/mass spectroscopy
- DMSO:
-
dimethylsulfoxide
- EDTA:
-
ethylenediaminetetraacetic acid
- DTPA:
-
diethylenetriaminepentaacetic acid
References
Wariishi, H., Valli, K., and Renganathan, V. (1989),J. Biol. Chem. 264, 14,185–14,191.
Juliette, L. Y., Hyman, M. R., and Arp, D. J. (1993),Appl. Environ. Microbiol. 59, 3718–3727.
Piette, L. H., Bulow, G., and Yamazaki, I. (1964),Biochim. Biophys. Acta 88, 120–129.
Kobayashi, S., Nakano, S., Goto, T., Kimura, T., and Schaap, A. P. (1986),Biochem. Biophys. Res. Commun. 135, 166–171.
Doerge, D. (1986),Arch. Biochem. Biophys. 244, 678–685.
Colonna, S., Gaggero, N., Manfredi, A., Casella, L., Gullotti, M., Carrea, G., and Pasta, P. (1990),Biochemistry 29, 10,465–10,468.
Casella, L., Gullotti, M., Ghezzi, R., Poli, S., Beringhelli, T., Colonna, S., and Carrea, G. (1992),Biochemistry 31, 9451–9459.
Klyachko, N. L. and Klibanov, A. M. (1992),Appl. Biochem. Biotechnol. 37, 53–68.
Dunford, H. B. (1991), inPeroxidases in Chemistry and Biology, vol. 2, Everse, J., Everse, K. E., and Grisham, M. B., eds., CRC, Boca Raton, pp. 1–24.
Keilin, D. and Hartree, E. F. (1950),Nature 166, 513–514.
King, N. K. and Winfield, M. E. (1963),J. Biol. Chem. 238, 1520–1528.
Ohlsson, P. I., and Paul, K. G. (1976),Acta Chem. Scand. B30, 373–375.
Kelder, P. P., de Mol, N. J., and Janssen, L. H. L. (1989),Biochem. Pharmacol. 38, 3593–3599.
Nelson, D. P. and Kiesow, L. A. (1972),Anal. Biochem. 49, 474–478.
Faison, B. D., Clark, T. M., Lewis, S. N., Ma, C. Y., Sharkey, D. M., and Woodward, C. A. (1991),Appl. Biochem. Biotechnol. 28/29, 237–251.
Rachmilewitz, E. A., Peisach, J., and Blumberg, W. E. (1971),J. Biol. Chem. 246, 3356–3366.
Giulivi, C. and Cadenas, E. (1993),FEBS 332, 287–290.
Perez, U. and Dunford, H. B. (1990),Biochim. Biophys. Acta 1038, 98–104.
Gutteridge, J. M. C., Richmond, R., and Halliwell, B. (1979),Biochem. J. 184, 469–472.
Gutteridge, J. M. C. (1986),FEBS Lett. 201, 291–295.
Puppo, A. and Halliwell, B. (1988),Biochem. J. 249, 185–190.
Arnao, M. B., Acosta, M., Del Rio, J. A., Varón, R., and Garcia-Cánovas, F. (1990),Biochim. Biophys. Acta 1041, 43–47.
Vincent, S. H., Grady, R. W., Shaklai, N., Snider, J. M., and Muller-Eberhard, U. (1988),Arch. Biochem. Biophys. 265, 539–550.
Beaven, G. H., d’Albis, A., and Gratzer, W. B. (1974),Eur. J. Biochem. 41, 539–546.
Shin, W. S., Yamashita, H., and Hirose, M. (1994),Biochem. J. 304, 81–86.
Rice, R. H., Lee, Y. M., and Brown, W. D. (1983),Arch. Biochem. Biophys. 221, 417–427.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Stachyra, T., Guillochom, D., Pulvin, S. et al. Hemoglobin, horseradish peroxidase, and heme-bovine serum albumin as biocatalyst for the oxidation of dibenzothiophene. Appl Biochem Biotechnol 59, 231–243 (1996). https://doi.org/10.1007/BF02783567
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02783567