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 V_max and K_M 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.