Abstract
Kinetic and physicochemical properties of Moringa oleifera peroxidase purified using a novel and cost efficient protocol was investigated with a view to providing information on its possible biotechnological potentials.Moringa oleifera peroxidase was purified to homogeneity in two steps, involving ATPS and size exclusion chromatography on Sephadex G-100 with a yield of 84.12 %. In-gel activity staining revealed the presence of one isoform of peroxidase. The purified peroxidase is monomeric with native and subunits molecular weight of 38.9 and 43.5 kDa respectively. Kinetic parameters - Vmax, Km(app)o-dianisidine, Km(app)H2O2 of the purified enzyme were 2.5 units/mg protein, 0.020 ± 0.04 mM and 1.37 ± 0.18 mM respectively. Its optimum pH and temperature were 5 and 30 °C respectively. The purified enzyme cross-linked BSA into an insoluble matrix with the aid of caffeic acid.The study concluded that the purification scheme adopted is rapid and efficient, the purified enzyme exhibited some physiochemical properties that make it suitable for various biotechnological applications.
Highlights
Peroxidases (EC 1.11.1.x) are a group of enzymes that contain heme, and they oxidize varieties of xenobiotics by using hydrogen peroxide (Saunders, 1973)
We reported in this study a rapid and efficient purification scheme for peroxidase from Moringa oleifera and demonstrated its potential in the synthesis of cross-link protein network and as a reporter enzyme
The purified peroxidase was stable over a pH range of 4.0–6.0 with its optimum pH around 5.0 and unstable at alkaline pH
Summary
Peroxidases (EC 1.11.1.x) are a group of enzymes that contain heme, and they oxidize varieties of xenobiotics by using hydrogen peroxide (Saunders, 1973). Peroxidase is an anti-oxidative enzyme that is widely distributed in microbe, plant, and animal tissue They represent a large group of heme-containing enzymes family (Van Huystee and Cairns, 1982). Other functions of peroxidase include their involvement in the biotransformation of drugs, chemicals, and polymers (Sakai et al, 2014) Their ability to oxidize phenolic has increased its use for various industrial applications, of which the most important ones include decolorization of waste (Lai and Lin, 2005; Dalal and Gupta, 2007; Jadhav et al, 2009), synthesis of various aromatic chemicals, removal of peroxides from foodstuff, industrial wastes and crosslinking of macromolecules (proteins) (Saitou et al 1991; Kim and Yoo, 1996) and in the biological field, as diagnostic kits for enzyme immunoassays and as an important component of ELISA system (Leon et al, 2002; Deepa and Arumughan, 2002)
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