Comparison of catalase activity in different organs of the potato (Solanum tuberosum L.) cultivars grown under field condition and purification by three-phase partitioning
Catalase (CAT) (H2O2: H2O2 oxidoreductase; EC 188.8.131.52) is capable of directly dismutating moderately reactive H2O2 into H2O and O2, and is regarded as one of the major enzymatic antioxidants in plants. The CAT isozymes are known to be differentially expressed and regulated. In this study, micropropagated potato plantlets of seven cultivars were grown under field condition with uniform agricultural practices. CAT activities were measured in the crude extracts from different potato organs namely tubers, leaves and stems at different stages of their growth. Relatively higher CAT activity was noticed in the very small actively growing tubers as compared to the other tissues. Cultivar-dependent differences were noticed in terms of the CAT activities which clearly indicated variation with regard to their antioxidative capacities. pH profile, thermostability and storage stability of CAT were examined. A simple and rapid three-phase partitioning (TPP) method worked effectively with regard to purification of CAT from the crude extracts. Both denaturing and non-denaturing PAGE analyses suggested that both the tuber-type and leaf-type CAT are tetrameric in nature and varied in size, possibly referred to the distinct isoforms in potato.
KeywordsPotato (Solanum tuberosum L.) cultivars Field condition Catalase (CAT) Three-phase partitioning (TPP) PAGE analyses
We gracefully thank UGC, Government of India, for providing UGC:MAN Fellowship to G. Kaur.
Compliance with ethical standards
Conflict of interest
The authors declare no conflict of interest.
Human and animal rights statement
This study did not involve human participants and/or animals.
- Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126Google Scholar
- Beaumont F, Jouve H-M, Gagnon J, Gaillard J, Pelmont J (1990) Purification and properties of a catalase from potato tubers (Solanum tuberosum). Plant Sci 72:19–26Google Scholar
- Boguszewska D, Grudkowska M, Zagdańska B (2010) Drought-responsive antioxidant enzymes in potato (Solanum tuberosum L.). Pot Res 53:373–382Google Scholar
- Demagante AL, Vander Zaag P (1988) The response of potato (Solanum spp.) to photoperiod and light intensity under high temperatures. Pot Res 31:73–83Google Scholar
- Dennison C, Lovrien R (1997) Three phase partitioning: concentration and purification of proteins. Protein Expres Purif 11:149–161Google Scholar
- Kandukuri SS, Noor A, Ranjini SS, Vijayalakshmi MA (2012) Purification and characterization of catalase from sprouted black gram (Vigna mungo) seeds. J Chromatogr B 889–890:50–54Google Scholar
- Kiss É, Szamos J, Tamás B, Borbás R (1998) Interfacial behavior of proteins in three-phase partitioning using salt-containing water/tert-butanol systems. Colloids Surf A Physicochem Eng Aspects 142:295–302Google Scholar
- Kumar V, Luthra SK, Bhardwaj V, Singh BP (2014) Indian potato varieties and their salient features. CPRI Technical Bulletin No. 78 (revised), ICAR-Central Potato Research Institute, Shimla, Himachal Pradesh, IndiaGoogle Scholar
- Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275Google Scholar
- Mhamdi A, Queval G, Chaouch S, Vanderauwera S, Breusegem FV, Noctor G (2010) Catalase function in plants: a focus on Arabidopsis mutants as stress-mimic models. J Expt Bot 61:4197–4220Google Scholar
- Özer B, Akardere E, Çelem EB, Önal S (2010) Three-phase partitioning as a rapid and efficient method for purification of invertase from tomato. Biochem Eng J 50:110–115Google Scholar
- Rahnama H, Ebrahimzadeh H (2005) The effect of NaCl on antioxidant enzyme activities in potato seedlings. Biol Plantarum 49:93–97Google Scholar
- Rojas-Beltran JA, Dejaeghere F, Abd Alla Kotb M, Du Jardin P (2000) Expression and activity of antioxidant enzymes during potato tuber dormancy. Pot Res 43:383–393Google Scholar
- Saxena L, Iyer BK, Ananthanarayan L (2007) Three phase partitioning as a novel method for purification of ragi (Eleusine coracana) bifunctional amylase/protease inhibitor. Process Biochem 42:491–495Google Scholar
- Scandalios JG, Guan L, Polidoros AN (1997) Catalases in plants: gene structure, properties, regulation, and expression. In: Scandalios JG (ed), Oxidative stress and the molecular biology of antioxidant defenses. Cold Spring Harbor Liboratory Press pp 343–406Google Scholar