Purification, characterization and selected inhibition properties of peroxidase from haricot bean (Phaseolus vulgaris L.)

Figures & data

Table 1. Purification of peroxidase from haricot bean (Phaseolus vulgaris L.).

Purification steps	Total enzyme activity(EU/mL.min)	Total protein(mg/mL)	Specific activity(EU/mg)	Purification fold
Homogenate	0.897	0.0305	29.40	1.0
(NH4)2SO4 precipitation and dialysis	0.715	0.0159	44.96	1.53
Ion-exchange chromatography	0.318	0.0098	32.44	1.09
Gel filtration chromatography	0.043	0.0017	25.29	0.86

Table 2. The inhibition effects of different chemicals on peroxidase activity from haricot bean (Phaseolus vulgaris L.).

Inhibitors	Concentration (mM)	Inhibition (%)
Citric acid	5.0	81
EDTA	20.0	90
CTAB	20.0	91
Sodium azide	5.0	99

Figure 1. Purification of peroxidase (POD) purified from haricot bean (Phaseolus vulgaris L.). (A) Ammonium sulphate precipitation, (B) CM-Sephadex exchange chromatography, and (C) gel filtration chromatography.

Figure 2. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) zymograme of peroxidase (POD) purified from haricot bean (Phaseolus vulgaris L.). Molecular weight markers (Lane 1) and gel filtration peak (Lane 2).

Figure 3. The effect of different conditions on peroxidase (POD) purified from haricot bean (Phaseolus vulgaris L.). (A) Optimum pH of POD, (B) the enzyme stability at different pH with time, (C) the effect of temperature on POD activity, (D) thermal stability of POD, and (E) the effect of ionic strength on POD.

Figure 4. Lineweaver–Burk plots for both substrate of guaiacol (A) and H₂O₂ (B) for POD purified from haricot bean (Phaseolus vulgaris L.).