Cyanobacterium Oscillatoria sp. peroxidase active at alkaline pH and high stability under chemical stresses

Main Article Content

Surasak Laloknam
Chaiyasard Kachensuwan


Cyanobacterial peroxidase enzyme extracted from Oscillatoria sp. SWU121 was purified from crude extract by 20-80% ammonium sulfate precipitation, DEAE cellulose ion-exchange chromatography, and Sephadex G-100 size exclusion chromatography. The purified Oscillatoria sp. peroxidase (OsPOX) exhibited a specific activity of 6106.63, while purification fold and yield were 17.45 and 34.70%, respectively. The OsPOX showed single band of protein on native and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE). The molecular weight was 45 kD similarly determined by gel filtration and SDS-PAGE suggesting that the OsPOX contained only one subunit. The apparent Km and Vmax values of the enzyme against phenol were 930 mM and 8078.25 protein-1, respectively. The temperature and pH optimum for OsPOX were 30ºC and pH 9.0, respectively. However, it was stable at 10 – 70ºC and pH 7.0 - 11.0. The presence of metal ions such as Na+ , Mn2+and Fe3+ enhanced peroxidase activity. On the other hand, Zn+ , Cd2+ and Hg2+ strongly inhibited the enzyme activity. SDS and EDTA reduced the peroxidase activity at 10 mM (20%). The OsPOX was found to be stable in the presence of urea. The affinity of the enzyme was highest for gallic acid, followed by ascorbic acid, phenol and cafeic acid. This finding showed that OsPOX was active at alkaline pH and stable in presence of urea.

Article Details



Ajila, C. M. and Prasada, Rao U. J. S. 2008. Purification and characterization of black gram (Vigna mungo) husk peroxidase. Journal of Molecular Catalysis B: Enzymatic 60, 36–44.

Al–Senaidy, A. and Ismael, M. (2011). Purification and characterization of membrane – bound peroxidase from date palm leaves (Phoenix dactylifera L.). Saudi Journal of Biological Sciences 18(3), 293–298.

Becana, M. and Lotassa, C. 2007. Oxidative stress as a response to salinity and aluminum. Lotus Newsletter 37(3), 98.

Bertelsmann, H., Kuehbacher, M., Weseloh, G., Kyriakopoulos, A. and Behne, D. 2007. Sperm nuclei glutathione peroxidases and their occurrence in animal species with cysteine-containing protamine. Biochimica et Biophysica Acta 1770, 1459–1467.

Bradford, M. 1976. A rapid method for the quantification of microgram quantities of proteins utilizing the principle of protein–dye binding. Analytical Biochemistry 72, 248–254.

Chen, M., Yao, S., Zhang, H. and Liang, X. 2010. Purification and characterization of a versatile peroxidase from edible mushroom Pleurotus eryngii. Chinese Journal of Chemical Engineering 18(5), 824–829.

Fatima, A., Husain, Q. and Khan, R. H. 2007. A peroxidase from bitter gourd (Momordica charantia): A comparison with horseradish peroxidase. Journal of Molecular Catalysis B: Enzymatic 47, 66–71.

Grigorieva, G. and Shestakov, S. 1982. Transformation in the cyanobacterium Synechocystis 6803, FEMS Microbiology Letter 13, 367–370.

Heinfing, A., Martènez, M. J., Martènez, A. T., Bergbauer, M. and Szewzyk, U. 1998. Purification and characterization of peroxidases from the dye-decolorizing fungus Bjerkandera adusta, FEMS Microbiology Letter 165, 43–50.

Ihsan, G. 2008. The effect of heavy metals on peroxidase from Jerusalem artichoke (Helianthus tuberosus L.) tubers. African Journal of Biotechnology 7(13), 2248–2253.

Incharoensakdi, A. and Laloknam, S. 2005. Nitrate uptake in the halotolerant cyanobacterium Aphanothece halophytica is energy-dependent driven by pH. Journal of biochemistry and molecular biology 38(4), 468–473.

Kachensuwan, C., Kongwithtaya, S., Phornphisutthimas, S. and Laloknam, S. 2012. A Screening of cyanobacteria producing the peroxidase. Journal of Research Unit on Science, Technology and Environment for Learning 3(1), 1–7. (in Thai)

Kavitha, K., Venkateraman, G. and Parida, A. 2008. An oxidative and salinity stress induced peroxisomal ascorbate peroxidase from Avicennia marina: Molecular and functional characterization. Plant Physiology and Biochemistry 46, 794–804.

Kongwithtaya, S., Laloknam, S. and Chairote, G. 2010. Purification and Characterization of Plant Peroxidase from Coccinia grandis (Ivy Gourd). Journal of Chemistry and Chemical Engineering 4(10), 13–19.

Kumar, S. and Sarla, S. M. 2008. Partial purification of superoxide dismutase and peroxidase from ber (Zizyphus mauritiana Lamk.) fruit using anion exchange chromatography. Physiology and Molecular Biology of Plants 14(3), 167–172.

Laloknam, S., Kanchitanurak, P., Boonburapong B., Rai, V. and Kongwitthaya, S. 2014. Inorganic and organic compounds of freshwater filamentous cyanobacteria under normal and salt stress conditions. Journal of Chemistry and Chemical Engineering 8, 1059–1067.

Melda, S., Ilhami, G., Murat, C., Ali, A., Hilal, M., Habibe, B. and Hasan, O. 2010. Purification and characterization of peroxidase from Turkish black radish (Raphanus sativus L.). Journal of Medicinal Plants Research 4(12), 1187–1196.

Miranda, M. V., Magri, M. L., Navarro, A. A. and del Canizo, O. 2002. Cascone, Study of variables involved in horseradish and soybean peroxidase purification by affinity chromatography on concanavalin A-Agarose. Process Biochemistry 38, 537–543.

Nelson, R. E., Fessler, L., Takagi, Y., Blumberg, B., Keene, D. R., Olson, P. F., Parker, C. G. and Fessler, J. H. 1994. Peroxidasin: a novel enzyme-matrix protein of Drosophila development, EMBO Journal 13, 3438–3447.

Ogawa, J., Triarsi, W., Sulistyaningdyah, T., Li, Q., Tanaka, H., Xie, S., Kana, K., Ikeda, T. and Shimizu, S. 2004. Two extracellular proteins with alkaline peroxidase activity, a novel cytochrome C and a cata-lase–peroxidase, from Bacillus sp. No.13. Bio - chimica et Biophysica Acta 1699, 65–75.

Rai, A. N., Söderbäck, E. and Bergman, B. 2000. Cyano - bacterium-plant symbioses. New Phytologist 147, 449–481.

Regelsberger, G. Jakopitsch, C., Plasser, L., Schwaiger, H., Furtmuller, P. Peschek, G., Zamocky, M. and Obin - ger, C. 2002. Occurrence and biochemistry of hy - doperoxidases in oxygenic phototrophic prokaryotes (Cyanobacteria). Plant Physiology and Biochemis - try 40, 479–490.

Sakamoto, M. and Komagata, K. 1996. Aerobic Growth of and Activities of NADH Oxidase and NADH Per - oxidase in Lactic Acid Bacteria. Journal of Fermen - tation and Bioengineering 82, 210–216.

Shannon, L., Kay, E. and Lew, J. 1966. Peroxidase isozymes from horseradish roots. I. isolation and physical properties. Journal of Biological Chemistry 241, 2166–2172.

Singh, J., Dubey, A., Diwakar, S., Rawat, S., Batra, N. and Joshi, A. 2010. Biochemical Characterization of peroxidase from the fruits of Mullus pumilus. Inter - national Research Journal of Biotechnology 1(4), 51–58.

Stanier, R. Y. and Cohen, B. G. 1977. Phototrophic prokar - yotes: the cyanobacteria. Annual Review of Micro - biology 31, 225–274.

Takeda, T., Yoshimura, K. and Shigeoka, S. 1998. Purifica - tion and characterization of ascorbate peroxidase in Chlorella vulgaris. Biochimie. 80, 295–301.

Veda, P. and Dwivedi, N. 2011. Purification and charac - terization of peroxidase from Leucaena leucoceph - ala, a tree legume. Journal of Molecular Catalysis B: Enzymatic 68, 168–173.

Vernwal, S., Yadav, R. and Yadav, K. 2006. Purification of a peroxidase from Solanum melongena fruit juice. Indian Journal of Biochemistry and Biophysics 43, 239–243.

Vitali, A., Botta, B., Monache, G. D., Zappitelli, S., Ric - ciardi, P., Melino, S., Petruzzelli, R. and Giardina, B. 1998. Purification and partial characterization of a peroxidase from plant cell cultures of Cassia didymobotrya and biotransformation studies. Bio - chemical Journal 331, 513–519.

Welinder, K. G. 1992. Superfamily of plant, fungal and bacterial peroxidases, Current Opinion in Structural Biology 2, 388–393.

Wright, H. and Nicell, J. A. 1999. Characterization of soybean peroxidase for the treatment of aqueous phenol. Bioresource Technology 70, 69–79.

Zamocky, M. 2004. Phylogenetic relationships in class I of the superfamily of bacterial, fungal, and plant peroxidases. European Journal of Biochemistry 271, 3297–3309.

Zheng, Z., Wang, Z. and Lin, R. 1999. Purification and some properties of a peroxidase from Tartary buck - wheat bran. Fagopyum16, 57–60.