Purification and characterization of alkaline-thermostable protease enzyme from pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme
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 electr...
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| Format: | Article |
| Language: | English |
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Hindawi Publishing Corporation
2014
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| Online Access: | http://psasir.upm.edu.my/id/eprint/35116/ http://psasir.upm.edu.my/id/eprint/35116/1/35116.pdf |
| _version_ | 1848847964481519616 |
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| author | Amid, Mehrnoush Abd Manap, Mohd Yazid Ahmad Zohdi, Nor Khanani |
| author_facet | Amid, Mehrnoush Abd Manap, Mohd Yazid Ahmad Zohdi, Nor Khanani |
| author_sort | Amid, Mehrnoush |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-15T09:26:58Z |
| format | Article |
| id | upm-35116 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T09:26:58Z |
| publishDate | 2014 |
| publisher | Hindawi Publishing Corporation |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-351162016-12-23T08:06:20Z http://psasir.upm.edu.my/id/eprint/35116/ Purification and characterization of alkaline-thermostable protease enzyme from pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme Amid, Mehrnoush Abd Manap, Mohd Yazid Ahmad Zohdi, Nor Khanani 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. Hindawi Publishing Corporation 2014 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/35116/1/35116.pdf 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 http://www.hindawi.com/journals/bmri/2014/259238/abs/ 10.1155/2014/259238 |
| spellingShingle | Amid, Mehrnoush Abd Manap, Mohd Yazid Ahmad Zohdi, Nor Khanani Purification and characterization of alkaline-thermostable protease enzyme from pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme |
| title | Purification and characterization of alkaline-thermostable protease enzyme from pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme |
| title_full | Purification and characterization of alkaline-thermostable protease enzyme from pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme |
| title_fullStr | Purification and characterization of alkaline-thermostable protease enzyme from pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme |
| title_full_unstemmed | Purification and characterization of alkaline-thermostable protease enzyme from pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme |
| title_short | Purification and characterization of alkaline-thermostable protease enzyme from pitaya (Hylocereus polyrhizus) waste: a potential low cost of the enzyme |
| title_sort | purification and characterization of alkaline-thermostable protease enzyme from pitaya (hylocereus polyrhizus) waste: a potential low cost of the enzyme |
| url | http://psasir.upm.edu.my/id/eprint/35116/ http://psasir.upm.edu.my/id/eprint/35116/ http://psasir.upm.edu.my/id/eprint/35116/ http://psasir.upm.edu.my/id/eprint/35116/1/35116.pdf |