Purification and selected biochemical properties of peroxidase from cress (Lepidium sativum sub sp. sativum)

Figures & data

Table 1. Purification steps of peroxidase from cress (Lepidium sativum sub sp. sativum).

Purification steps	Total volume (mL)	Enzyme activity (EU/mL)	Total enzyme activity (EU)	Protein (mg)	Total protein (mg)	Specific activity (EU/mg protein)	Yield (%)	Purification fold
Homogenate	50	1038	51900	1.34	67.0	774.63	100.0	1.0
(NH4)2SO4 precipitation	12	1573	18876	1.066	12.8	1474.68	36.4	1.9
Gel filtration chromatography	3	708	2124	0.170	0.51	4164.70	4.1	5.4
Ion Exchange chromatography	3	104	316	0.0029	0.0087	36321.83	0.61	46.9

Figure 1. Purification of peroxidase (cPOD) from cress (Lepidium sativum sub sp. sativum). (A) Ammonium sulphate precipitation, (B) Gel filtration chromatography, (C) CM-Sephadex ion exchange chromatography.

Figure 2. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) zymogram of peroxidase (cPOD) purified from cress (Lepidium sativum sub sp. sativum). Ion exchange chromatography peak (Lane 1) and molecular weight markers (Lane 2).

Figure 3. The effect of different conditions on peroxidase (cPOD) purified from cress (Lepidium sativum sub sp. sativum). (A) Optimum temperature of cPOD, (B) optimum pH of cPOD, (C) the effect of ionic strength on cPOD, (D) Stabile pH for cPOD.

Figure 4. Lineweaver-Burk graphs substrate pattern of peroxidase from cress (Lepidium sativum sub sp. sativum). (A) Guaiacol substrate, (B) H₂O₂ substrate.

Table 2. Inhibition properties of some inhibitors on cPOD activity.

Metal ions	Concentration (mM)	IC50 (mM)	Ki (mM)	Inhibition type
Al^3+	0–30	8.36	5.78	Noncompetitive
Cu^2+	0–30	31.56	20.90	Noncompetitive
Cr^3+	0–30	20.71	10.30	Noncompetitive
CTAB	0–2.5	1.98	1.160	Competitive