Purification and Thermal Characterization of a Novel Peroxidase from a Local Chick Pea Cultivar

Title: Purification and Thermal Characterization of a Novel Peroxidase from a Local Chick Pea Cultivar

Volume: 13 Issue: 8

Author(s): H. N. Bhatti, A. Najma, M. Asgher, M. A. Hanif and M. A. Zia

Affiliation:

Keywords: Chick pea peroxidase, Cicer arietinum L, thermal denaturation, purification, thermostability, free energy

Abstract: A novel peroxidase isolated from a local chick pea (Cicer arietinum L.) cultivar (Balksar 2000) was purified by means of ammonium sulfate precipitation, DEAE-cellulose chromatography and two runs on gel filtration. The purified enzyme has a specific activity of 2045 U/mg with 17 % activity recovery. The molecular mass of the enzyme was estimated to be 39 kDa by SDS-polyacrylamide gel electrophoresis. Optimum pH and temperature of the enzyme were 5.5 and 45°C respectively. The thermal denaturation of local chick pea peroxidase was studied in aqueous solution at temperatures ranging from 45°C to 65°C. The temperature of 50% inactivation of the enzyme was found to be 68°C. The enthalpy (ΔH*) and free energy (ΔG*) of thermal denaturation of chick pea peroxidase were 101.4 and 103.4 k J/mol respectively at 65°C.Metals like Zn2+, Mn2+ , Hg2+, Co2+ and Al3+ slightly inhibited the peroxidase activity while Ca2+, Mg2+ and Ba2+ have no effect on enzyme activity. The high specific activity and thermal stability make chick pea peroxidase an alternative to horseradish peroxidase (HRP) in various applications.

Cite this article as:

Bhatti N. H., Najma A., Asgher M., Hanif A. M. and Zia A. M., Purification and Thermal Characterization of a Novel Peroxidase from a Local Chick Pea Cultivar, Protein & Peptide Letters 2006; 13 (8) . https://dx.doi.org/10.2174/092986606777841271