An efficient methodology for the purification of date palm peroxidase: Stability comparison with horseradish peroxidase (HRP)

Abstract

In the present study, Peroxidase from date palm ( Phoenix dactylifera ) leaves was purified to homogeneity by three-step procedure including aqueous two-phase system, hydrophobic and Ion-exchange chromatography. The enzyme migrated as single band on SDS-PAGE giving molecular weight of 68 ± 3 kDa. The purification factor for purified date palm peroxidase was 68 with high 41% yield. Enzymatic assays together with far-UV circular dichroism (CD), intrinsic and extrinsic fluorescence studies were carried out to monitor the structural stability of date palm and horseradish peroxidase (HRP) against various pH and temperatures. Activity measurements illustrated different pH stability for date palm and HRP. Both peroxidases are more susceptible to extreme acidic conditions as suggested by 4 & 15 nm red shift in date palm and HRP, respectively. Secondary structure analysis using far UV-CD exhibited predominance of α-helical (43.8%) structure. Also, pH induces loss in the secondary structure of date palm peroxidase. Thermal stability analysis revealed date palm peroxidase is more stable in comparison to HRP. In summary, date palm peroxidases could be promising enzymes for various applications where extreme pH and temperature is required.

Purification and characterization of peroxidase from date palm cv. Agwa fruits

“…The different molecular weights of 55 and 68 kDa were reported for peroxidases from date palm leaves. [11,16] The similar molecular weight was detected for peroxidase from Raphanus sativus (44 kDa). [31] The pH optimum of the enzyme is very important for enzyme activity, where the active site changed by changing the ionic state of amino acids of enzyme.…”

A novel peroxidase from runner bean ( Phaseolus coccineus L.): Enhanced affinity purification, characterization, and dye decolorization activity

“…Calotropis procera leaves 2-fold (Mafulul et al, 2018) and from date palm 68-fold with a yield of 41% (Al-Bagmi et al, 2019). Although the PODs can be purified by multi-step methods (ammonium sulfate precipitation, acetone precipitation, ion exchange, and size-exclusion chromatography), these complicated steps lead to the loss of yield, fold, and time.…”

Immobilization of Low-Cost Alternative Vegetable Peroxidase (Raphanus sativus L. peroxidase): Choice of Support/Technique and Characterization

“…Peroxidases (EC 1.11.1.7) are enzymes that catalyze the oxidation of a wide variety of substrates dependent on H 2 O 2 [1]. Among vegetable peroxidases, horseradish peroxidase (HRP) is the most studied [2,3] due to the numerous possibilities of applications, such as bioremediation, biosensors and diagnostic kits [4,5].…”