African Journal of
Agricultural Research

  • Abbreviation: Afr. J. Agric. Res.
  • Language: English
  • ISSN: 1991-637X
  • DOI: 10.5897/AJAR
  • Start Year: 2006
  • Published Articles: 6574

Full Length Research Paper

Purification and characterisation of a plant peroxidase from rocket (Eruca vesicaria sbsp. Sativa) (Mill.) (syn. E. sativa) and effects of some chemicals on peroxidase activity

Hayrunnisa Nadaroglu1*, Neslihan Celebi1, Nazan Demir2 and Yasar Demir2
1Department of Food Technology, Erzurum Vocational Training School, Ataturk University, 25240 Erzurum, Turkey. 2Department of Chemistry, Science Faculty, Mugla University, 48000 Mugla, Turkey.
Email: [email protected]

  •  Accepted: 20 May 2013
  •  Published: 06 June 2013

Abstract

Rocket (Eruca vesicaria sbsp. Sativa) (Mill.) (syn. E. sativa) was grown and used widely in Turkey as a garnish in salads. A peroxidase (POD) from leaves of rocket (Eruca vesicaria sbsp. Sativa) was purified using sequential (NH4)2SO4 precipitation, CM-Sephadex and Sephacryl S-200 chromatographies. A peroxidase (POD) was purified 220.3-fold from the Rocket (E. vesicaria sbsp. Sativa) with an overall yield of 80.79%. The purified enzyme has an optimum pH, 6.0 and its optimum temperature was 40°C. The Vmax and KM values were determined by Lineweaver-Burk graphics using different substrates. The purification degree and the molecular mass of the enzyme (34 kDa) were determined by SDS-PAGE and gel filtration chromatography. POD enzyme activity was strongly inhibited by Ca2+, Mn2+, Hg22+, Zn2+ and Fe2+ as metal ions and SDS, EDTA, ascorbic acid, dithioeritritol as chemicals. But, Ni2+, Co2+, Cu2+ slightly activated the enzyme. They inhibited in the different range of peroxidase activity. Changes of POD enzyme’s kinetic parameters were most important during chemicals and metal ions metabolism, because they were risk for environmental pollution.

 

Key words: Rocket (Eruca vesicaria sbsp. Sativa), peroxidase (POD), purification, metal ions.