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


Citation

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

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.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Food Science and Technology
DOI Number: https://doi.org/10.1155/2014/259238
Publisher: Hindawi Publishing Corporation
Keywords: Alkaline-thermostable protease enzyme; Pitaya waste; Hylocereus polyrhizus; Low cost; Enzyme
Depositing User: Nurul Ainie Mokhtar
Date Deposited: 30 Dec 2015 10:57
Last Modified: 23 Dec 2016 08:06
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1155/2014/259238
URI: http://psasir.upm.edu.my/id/eprint/35116
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