Purification of an infection-related acidic peroxidase from pearl millet seedlings

Devaiah, Shivakumar Pattada and Shetty, Shekar H. (2009) Purification of an infection-related acidic peroxidase from pearl millet seedlings. Pesticide Biochemistry and Physiology, 94 (2-3). pp. 119-126. ISSN 1095-9939

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Official URL: http://doi.org/10.1016/j.pestbp.2009.04.010

Abstract

Higher basal level of peroxidase activity was observed in highly resistant pearl millet cultivar IP 18292. Upon inoculation with downy mildew pathogen, Sclerospora graminicola, up to 60% increase in peroxidase activity was observed in highly resistant seedlings over the period of time. Iso-electric focusing analysis revealed that, two acidic isozymes of peroxidase with the pI of 5.9 and 5.1 present only in IP 18292 pearl millet seedlings. Upon inoculation with downy mildew pathogen, accumulation of these to isozymes was increased. These results indicated the possible involvement of acidic peroxidase in pearl millet defense. To study the nature of the acidic peroxidase which increases upon inoculation was purified from seedlings of highly resistant pearl millet cultivar using DEAE-Sepharose and Sephadex G-100 columns. The purified enzyme has a molecular weight of 21.8 kDa on SDS-PAGE and has a pI of 5.1. The optimum pH for maximum peroxidase activity was found to be at pH 7.0 and was resistant to high temperature (27-60 degrees C). The K-m for H2O2 and V-max of the enzyme reaction were 5.26 mM and 322.58 units, respectively. Purified peroxidase enzyme was found to be CaCl2 dependent and both MgCl2 and ZnCl2 showed inhibitory effect on enzyme activity. Sodium azide and EDTA inhibited the enzyme and EGTA found to be specific inhibitor of peroxidase. (C) 2009 Elsevier Inc. All rights reserved.

Item Type: Article
Uncontrolled Keywords: Pearl millet; Peroxidase; Purification; Sclerospora graminicola
Subjects: B Life Science > Biotechnology
Divisions: Department of > Biotechnology
Depositing User: Shrirekha N
Date Deposited: 26 Aug 2019 06:30
Last Modified: 26 Aug 2019 06:30
URI: http://eprints.uni-mysore.ac.in/id/eprint/7028

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