title: Purification and characterization of alkaline-thermostable protease enzyme from Pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme.
abstract: The thermoalkaline protease enzyme from pitaya (Hylocereus polyrhizus) waste was purified by a factor of 221.2 with 71.3% recovery using ammonium sulphate precipitation, gel filtration, and cation exchange chromatography. Gel filtration chromatography together with sodium dodecyl sulphate gel electrophoresis (SDS-PAGE) revealed that the enzyme is monomeric with a molecular weight of 26.7 kDa. The apparent K m and V max of the protease were 2.8 mg/mL and 31.20 u/min, respectively. The optimum pH and temperature were 8.0 and 70°C. The enzyme was highly active and stable over a wide pH range (from pH 3.0 to pH 11.0 with the optimum activity at pH 8.0). The protease has broad specificity toward azocasein, casein, hemoglobin, and gelatine. Activity of the enzyme was inhibited by Fe(2+) and Zn(2+), while protease activity was increased in the presence of Ca(2+) and Mg(2+) and Cu(2+) by factors of 125%, 110%, and 105%, respectively. The alkaline protease showed extreme stability toward surfactants and oxidizing agent. The purified protease exhibited extreme stability in the presence of organic solvents and inhibitors. In addition, the enzyme was relativity stable toward organic solvents and chelating agents, such as ethylenediaminetetraacetic acid (EDTA). The enzyme, derived from pitaya peel, possesses unique characteristics and could be used in various industrial and biotechnological applications.
authors.affiliation: Department of Food Technology, Faculty of Food Science and Technology, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor, Malaysia.
authors.forename: Mehrnoush
authors.identifier: 0000-0003-1517-2326
authors.initials: M
authors.lastname: Amid
authors.affiliation: Department of Food Technology, Faculty of Food Science and Technology, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor, Malaysia.
authors.forename: Mohd Yazid A B D
authors.initials: MY
authors.lastname: Manap
authors.affiliation: Department of Food Technology, Faculty of Food Science and Technology, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor, Malaysia.
authors.forename: Nor Khanani
authors.initials: NK
authors.lastname: Zohdi
mesh_terms: D000383:Agriculture; D029421:Cactaceae / Q000201:enzymology*; D002384:Catalysis; D004789:Enzyme Activation; D005638:Fruit / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007220:Industrial Waste / Q000032:analysis; D007700:Kinetics; D008296:Malaysia; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification*; D010936:Plant Extracts / Q000737:chemistry* / Q000302:isolation & purification*; D013696:Temperature
chemical_list: D007220:Industrial Waste; D010936:Plant Extracts; D010447:Peptide Hydrolases