Production and Purification of Peroxidase from Tissue Cultures of Cauliflower (Brassica oleracea L. var. Botrytis)

Year 2021, , 1121 - 1128, 30.10.2021

Züleyha Almaz , Şeyma Önlü , Aykut Öztekin

https://doi.org/10.16984/saufenbilder.844705

Cited By: 1

Abstract

The Plant peroxidases are remarkable enzymes due to their widespread use in industry. These enzymes, which are capable of catalyzing the oxidation of various organic and inorganic substrates, have been used in clinical diagnosis, detoxification reactions and organic synthesis. In this study, in vitro production and purification of peroxidase enzyme from cauliflower plant was proposed. Firstly, sterile seedlings were obtained from MS/B5 nutrient medium without growth regulator from cauliflower seeds and calluses from medium containing 0.5 mg / L 2.4-D. Callus and seedlings were powdered with liquid nitrogen and then homogenized. Peroxidase enzymes were purified from these homogenates using affinity technique. SDS-PAGE electrophoresis was performed to determine the molecular weight of the purified enzymes and single bands was observed at approximately 46 kDa. In addition, KM and Vmax values of the callus peroxidase enzyme were determined for guaiacol, pyrogallol and H2O2 substrates.

Keywords

cauliflower (brassica oleracea l. var. botrytis), plant tissue culture, peroxidase purification

References

• [1] V. P. Pandey, M. Awasthi, S. Singh, S. Tiwari, and U. N. Dwivedi, "A comprehensive review on function and application of plant peroxidases," Biochemistry and Analytical Biochemistry, vol. 6, no. 01, p. 308, 2017.
• [2] C. Regalado, B. E. García-Almendárez, and M. A. Duarte-Vázquez, "Biotechnological applications of peroxidases," Phytochemistry Reviews, vol. 3, no. 1-2, pp. 243-256, 2004.
• [3] F. Passardi, C. Cosio, C. Penel, and C. Dunand, "Peroxidases have more functions than a Swiss army knife," Plant cell reports, vol. 24, no. 5, pp. 255-265, 2005.
• [4] J. Jia, B. Wang, A. Wu, G. Cheng, Z. Li, and S. Dong, "A method to construct a thirdgeneration horseradish peroxidase biosensor: self-assembling gold nanoparticles to three-dimensional sol− gel network," Analytical Chemistry, vol. 74, no. 9, pp. 2217-2223, 2002.
• [5] E. Agostini, J. Hernández‐Ruiz, M. B. Arnao, S. R. Milrad, H. A. Tigier, and M. Acosta, "A peroxidase isoenzyme secreted by turnip (Brassica napus) hairy‐root cultures: inactivation by hydrogen peroxide and application in diagnostic kits," Biotechnology and applied biochemistry, vol. 35, no. 1, pp. 1-7, 2002.
• [6] A. Heller and M. S. Vreeke, "Soybean peroxidase electrochemical sensor," ed: Google Patents, 1997.
• [7] A. Bhunia, S. Durani, and P. P. Wangikar, "Horseradish peroxidase catalyzed degradation of industrially important dyes," Biotechnology and bioengineering, vol. 72, no. 5, pp. 562-567, 2001.
• [8] K. Tatsumi, S. Wada, and H. Ichikawa, "Removal of chlorophenols from wastewater by immobilized horseradish peroxidase," Biotechnology and Bioengineering, vol. 51, no. 1, pp. 126-130, 1996.
• [9] J. Liu, L. Ye, and Y. Weiping, "Copolymerization of lignin with cresol catalysed by peroxidase in reversed micellar systems," Electronic Journal of Biotechnology, vol. 2, no. 2, pp. 7-8, 1999.
• [10] M. Chivukula, J. T. Spadaro, and V. Renganathan, "Lignin peroxidase-catalyzed oxidation of sulfonated azo dyes generates novel sulfophenyl hydroperoxides," Biochemistry, vol. 34, no. 23, pp. 7765- 7772, 1995.
• [11] S.-S. Kwak, S.-K. Kim, I.-H. Park, and J. R. Liu, "Enhancement of peroxidase activity by stress-related chemicals in sweet potato," Phytochemistry, vol. 43, no. 3, pp. 565-568, 1996.
• [12] A. Cardinali et al., "Purification and characterization of a cationic peroxidase from artichoke leaves," Journal of the Science of Food and Agriculture, vol. 87, no. 7, pp. 1417-1423, 2007.
• [13] G. Fernández and A. B. Pomilio, "Optimized growth conditions and determination of the catalytic type of the peptidase complex from a novel callus culture of pineapple (Ananas comosus)," Mol Med Chem, vol. 1, pp. 39-49, 2003.
• [14] N. González-Rábade, M. del Carmen Oliver-Salvador, E. Salgado-Manjarrez, and J. A. Badillo-Corona, "In vitro production of plant peroxidases—a review," Applied biochemistry and biotechnology, vol. 166, no. 7, pp. 1644- 1660, 2012.
• [15] M. Parkinson, T. Cotter, and P. Dix, "Peroxidase production by cell suspension and hairy root cultures of horseradish (Armoracia rusticana)," Plant Science, vol. 66, no. 2, pp. 271-277, 1990.
• [16] P. Soudek, R. Podlipna, P. Marsik, and T. Vanek, "Optimalization of the peroxidase production by tissue cultures of horseradish in vitro," Biologia plantarum, vol. 49, no. 4, pp. 487-492, 2005.
• [17] E. Agostini, S. M. de Forchetti, and H. A. Tigier, "Production of peroxidases by hairy roots ofBrassica napus," Plant cell, tissue and organ culture, vol. 47, no. 2, pp. 177- 182, 1997.
• [18] R. Vazquez-Duhalt, "Peroxidase activity in calluses and cell suspension cultures of radishRaphanus sativus var. Cherry Bell," Plant cell, tissue and organ culture, vol. 18, no. 3, pp. 321-327, 1989.
• [19] A. Oztekin et al., "Purification of peroxidase enzyme from radish species in fast and high yield with affinity chromatography technique," Journal of Chromatography B, vol. 1114, pp. 86-92, 2019.
• [20] C. E. Bach et al., "Measuring phenol oxidase and peroxidase activities with pyrogallol, L-DOPA, and ABTS: effect of assay conditions and soil type," Soil Biology and Biochemistry, vol. 67, pp. 183-191, 2013.
• [21] M. M. Bradford, "A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding," Analytical biochemistry, vol. 72, no. 1-2, pp. 248-254, 1976.
• [22] U. K. Laemmli, "Cleavage of structural proteins during the assembly of the head of bacteriophage T4," nature, vol. 227, no. 5259, p. 680, 1970.
• [23] C. R. Merril, "[36] Gel-staining techniques," in Methods in enzymology, vol. 182: Elsevier, 1990, pp. 477-488.
• [24] Y. Hu, J. Wu, P. Luo, and Y. Mo, "Purification and partial characterization of peroxidase from lettuce stems," African Journal of Biotechnology, vol. 11, no. 11, pp. 2752-2756, 2012.
• [25] C. B. Lavery et al., "Purification of peroxidase from horseradish (Armoracia rusticana) roots," Journal of agricultural and food chemistry, vol. 58, no. 15, pp. 8471-8476, 2010.
• [26] E. Koksal and I. Gulcin, "Purification and characterization of peroxidase from cauliflower (Brassica oleracea L. var. botrytis) buds," Protein and Peptide Letters, vol. 15, no. 4, pp. 320-326, 2008.
• [27] T. Rudrappa, V. Lakshmanan, R. Kaunain, N. M. Singara, and B. Neelwarne, "Purification and characterization of an intracellular peroxidase from genetically transformed roots of red beet (Beta vulgaris L.)," Food chemistry, vol. 105, no. 3, pp. 1312-1320, 2007.