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Purification and characterization of Cu,Zn superoxide dismutase from pumpkin (Cucurbita moschata) pulp

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Abstract

Superoxide dismutase (SOD, EC 1.15.1.1) plays an important role in pharmaceutical, neutraceutical and cosmeceutical industries as it can eliminate toxic superoxide radicals. Copper and zinc SOD was purified from pumpkin (Cucurbita moschata) stringy pulp. The stepwise procedure consisted phosphate extraction, heat treatment, ammonium sulfate precipitation, Sephadex G-100 gel filtration and DEAE-52 chromatography. This procedure resulted in a single band on native polyacrylamide gels and sodium dodecyl sulfate–polyacrylamide gels. The enzyme was purified 54.4-fold with its specific activity of 1,794 U/mg protein and the yield of 23.1 %. The enzyme molecular mass was estimated at 35.3 ± 0.6 kDa by gel filtration. The subunit molecular mass of this homo-dimeric protein was estimated at 17.5 ± 0.3 kDa on sodium dodecyl sulfate–polyacrylamide gels. The purified enzyme, which was characterized as a H2O2-sensitive SOD, contained approximately one copper atom and one zinc atom per subunit, indicating that it belongs to the Cu,Zn-SOD. The enzyme was found to remain thermostable lower than 50 °C. Pumpkin pulp Cu,Zn-SOD appeared to remain wider range of pH stability than other plant Cu,Zn-SOD, as it showed a strong and steady activity at a wider range of pH from 4 to 12 at 25 °C for 2 h. The enzyme was slightly inhibited by SDS and NaN3, but was not inhibited by Na3BO3 and sodium citrate. Therefore, these results suggested that pumpkin stringy pulp which is considered as a waste can be used as a rich source of Cu,Zn-SOD and can be commercially exploited in cosmeceuticals, pharmaceuticals and neutraceuticals.

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References

  1. Paris HS, Brown RN (2005) The genes of pumpkin and squash. HortScience 40:1620–1630

    CAS  Google Scholar 

  2. Fu C, Shi H, Li Q (2006) A review on pharmacological activities and utilization technologies of pumpkin. Plant Foods Hum Nutr 61:73–80

    CAS  Google Scholar 

  3. Shi J, Yi C, Ye X, Xue S, Jiang Y, Ma Y, Liu D (2010) Effects of supercritical CO2 fluid parameters on chemical composition and yield of carotenoids extracted from pumpkin. LWT-Food Sci Technol 43:39–44

    Article  CAS  Google Scholar 

  4. Xanthopoulou MN, Nomikos T, Fragopoulou E, Antonopoulou S (2009) Antioxidant and lipoxygenase inhibitory activities of pumpkin seed extracts. Food Res Int 42:641–646

    Article  CAS  Google Scholar 

  5. Parker MW, Blake CC (1988) Iron- and manganese-containing superoxide dismutases can be distinguished by analysis of their primary structures. FEBS Lett 229:377–382

    Article  CAS  Google Scholar 

  6. Valdivia A, Pérez-Álvarez S, Aroca-Aguilar JD, Ikuta I, Jordán J (2009) Superoxide dismutases: a physiopharmacological update. J Physiol Biochem 65:195–208

    Article  CAS  Google Scholar 

  7. Moreira PI, Siedlak SL, Aliev G, Zhu X, Cash AD, Smith MA, Perry G (2005) Oxidative stress mechanisms and potential therapeutics in Alzheimer disease. J Neural Transm 112:921–932

    Article  CAS  Google Scholar 

  8. Lubrano V, Di Cecco P, Zucchelli GC (2006) Role of superoxide dismutase in vascular inflammation and in coronary artery disease. Clin Exp Med 6:84–88

    Article  CAS  Google Scholar 

  9. He N, Li Q, Sun D, Ling X (2008) Isolation, purification and characterization of superoxide dismutase from garlic. Biochem Eng J 38:33–38

    Article  CAS  Google Scholar 

  10. Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287

    Article  CAS  Google Scholar 

  11. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  12. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of Bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  13. Wang Z, He Z, Li S, Yuan Q (2005) Purification and partial characterization of Cu, Zn containing superoxide dismutase from entomogenous fungal species Cordyceps militaris. Enzyme Microb Technol 36:862–869

    Article  Google Scholar 

  14. Kochhar S, Kochhar VK (2008) Identification and characterization of a super-stable Cu–Zn SOD from leaves of turmeric (Curcuma longa L.). Planta 228:307–318

    Article  CAS  Google Scholar 

  15. Haddad NIA, Yuan Q (2005) Purification and some properties of Cu, Zn superoxide dismutase from Radix lethospermi seed, kind of Chinese traditional medicine. J Chromatogr B 818:123–131

    Article  CAS  Google Scholar 

  16. Ragusa S, Cambria MT, Scarpa M, Di Paolo ML, Falconi M, Rigo A, Cambria A (2001) Properties of purified cytosolic isoenzyme I of Cu, Zn-superoxide dismutase from Nicotiana plumbaginifolia leaves. Protein Expr Purif 23:261–269

    Article  CAS  Google Scholar 

  17. Babitha MP, Prakash HS, Shekar shetty H (2002) Purification and partial characterization of manganese superoxide dismutase from downy mildew resistant pearl millet seedlings. Plant Sci 163:917–924

    Article  CAS  Google Scholar 

  18. Vyas D, Kumar S (2005) Purification and partial characterization of a low temperature responsive Mn-SOD from tea (Camellia sinensis (L.) O. Kuntze). Biochem Biophys Res Commun 329:831–838

    Article  CAS  Google Scholar 

  19. Ahmed H, Schott EJ, Gauthier JD, Vasta GR (2003) Superoxide dismutases from the oyster parasite Perkinsus marinus: purification, biochemical characterization, and development of a plate microassay for activity. Anal Biochem 318:132–141

    Article  CAS  Google Scholar 

  20. Colla G, Rouphael Y, Cardarelli M, Salerno A, Rea A (2010) The effectiveness of grafting to improve alkalinity tolerance in watermelon. Environ Exp Bot 68:283–291

    Article  CAS  Google Scholar 

  21. Skarpanska-Stejnborn A, Pilaczynska-Szczesniak L, Basta P, Deskur-Smielecka E, Woitas-Slubowska D, Adach Z (2011) Effects of oral supplementation with plant superoxide dismutase extract on selected redox parameters and an inflammatory marker in a 2,000-m rowing-ergometer test. Int J Sport Nutr Exerc Metabol 21:124–134

    Google Scholar 

Download references

Acknowledgments

This work was supported by the open fund of Research Center of Green Manufacturing and Energy-Saving and Emission Reduction Technology in Wuhan University of Science and Technology (B1207).

Conflict of interest

Xiaorong Qin, Mingjin Zhang and Lin Wu declare that we have no conflict of interest.

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Authors and Affiliations

  1. College of Chemical Engineering and Technology, Wuhan University of Science and Technology, Wuhan, 430081, People’s Republic of China

    Xiaorong Qin & Lin Wu

  2. Wuhan Institute of Physics and Mathematics, The Chinese Academy of Sciences, Wuhan, 430071, People’s Republic of China

    Mingjin Zhang

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  1. Xiaorong Qin
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  2. Mingjin Zhang
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  3. Lin Wu
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Correspondence to Xiaorong Qin.

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Qin, X., Zhang, M. & Wu, L. Purification and characterization of Cu,Zn superoxide dismutase from pumpkin (Cucurbita moschata) pulp. Eur Food Res Technol 235, 1049–1054 (2012). https://doi.org/10.1007/s00217-012-1829-1

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  • DOI: https://doi.org/10.1007/s00217-012-1829-1

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