Purification, characterization, and inhibition sensitivity of peroxidase from wheat (Triticum aestivum ssp. vulgare)

Figures & data

Table 1. The results of purification of peroxidase from wheat (Triticum aestivum ssp. Vulgare).

Purification steps	Total enzyme activity (EU/mL.min)	Total protein (mg)	Specific activity (EU/mg)	Purification fold
Homogenate	0.022	1.152	0.02	1.0
(NH4)2SO4 precipitation and dialysis	0.903	0.892	1.01	51
Ion exchange chromatography	0.139	0.535	0.26	14
Sephadex-G-25	0.881	0.162	5.43	284

Table 2. The inhibition effects of different chemicals on peroxidase (wPOD) activity from wheat (Triticum aestivum ssp. Vulgare).

Inhibitors	Concentration (mM)	Inhibition (%)
Citric acid	5.0	94
EDTA	20.0	89
CTAB	20.0	95
Sodium azide	5.0	79

Figure 1. Purification of peroxidase (wPOD) purified from wheat (Triticum aestivum ssp. Vulgare). A: Ammonium sulphate precipitation; B: CM-Sephadex anion exchange chromatography; and C: Gel filtration chromatography.

Figure 2. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) zymograme of peroxidase (wPOD) purified from wheat (Triticum aestivum ssp. Vulgare). Molecular weight markers (Lane 1), and gel filtration peak (Lane 2–3).

Figure 3. The effect of different conditions on peroxidase (wPOD) purified from wheat (Triticum aestivum ssp. Vulgare). A: Optimum pH of wPOD; B: The enzyme stability at different pH with time; C: The effect of temperature on wPOD activity; D: Thermal stability of wPOD; and E: The effect of ionic strength on wPOD.

Figure 4. A: Lineweaver-Burk plots for both substrate of guaiacol; and B: H₂O₂ for peroxidase (wPOD) purified from wheat (Triticum aestivum ssp. Vulgare).