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Purification and characterization of alkaline-thermostable protease enzyme from pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme

Amid, Mehrnoush and Abd Manap, Mohd Yazid and Ahmad Zohdi, Nor Khanani (2014) Purification and characterization of alkaline-thermostable protease enzyme from pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme. BioMed Research International, 2014 . art. no. 259238. pp. 1-8. ISSN 2314-6133; ESSN: 2314-6141

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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 m a x 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 Fe2+ and Zn2+, while protease activity was increased in the presence of Ca2+ and Mg2+ and Cu2+ 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.

Item Type:Article
Keyword:Alkaline-thermostable protease enzyme; Pitaya waste; Hylocereus polyrhizus; Low cost; Enzyme
Faculty or Institute:Faculty of Food Science and Technology
Publisher:Hindawi Publishing Corporation
DOI Number:10.1155/2014/259238
Altmetrics:http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1155/2014/259238
ID Code:35116
Deposited By: Nurul Ainie Mokhtar
Deposited On:30 Dec 2015 18:57
Last Modified:23 Dec 2016 16:06

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