Isolation, purification and characterization of 5'-phosphodiesterase from Aspergillus fumigatus
| Autor: | Qiuxia Li, Yingying Fan, Zhiting Luo, Zongwen Pang, Bing Han, Yang Liu, Hua Qiu, Shubo Li |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Protein Conformation lcsh:Medicine 01 natural sciences Catalysis Aspergillus fumigatus 03 medical and health sciences Hydrolysis 010608 biotechnology Nucleotide lcsh:Science chemistry.chemical_classification Multidisciplinary biology Phosphoric Diester Hydrolases lcsh:R Active site Phosphodiesterase biology.organism_classification Enzyme assay Molecular Weight 030104 developmental biology Enzyme Biochemistry chemistry biology.protein Specific activity lcsh:Q Research Article |
| Zdroj: | PLoS ONE PLoS ONE, Vol 12, Iss 10, p e0186011 (2017) |
| ISSN: | 1932-6203 |
| Popis: | 5′-Phosphodiesterase (5′-PDE) catalyzes the hydrolysis of ribonucleic acid to obtain a mixture of ribonucleotides, such as 5′-guanosine monophosphate and 5′-adenosine monophosphate. In this study, a 5'-PDE was newly isolated and purified from Aspergillus fumigatus. Following purification, this enzyme exhibited a specific activity of 1036.76 U/mg protein, a molecular weight of 9.5 kDa, and an optimal temperature and pH for enzyme activity of 60°C and 5.0, respectively. However, its activity was partially inhibited by Fe3+, Cu2+, and Zn2+, but slightly improved by the presence of K+ and Na+. Additionally, chemical-modification experiments were also applied to investigate the structural information of 5'-PDE, in which the residues containing carboxyl and imidazole groups were essential for enzyme activity based on their localization in the 5′-PDE active site. Furthermore, purified 5′-PDE could specifically catalyze the synthesis of ribonucleotides with a Vmax 0.71 mmol/mg·min and a KM of 13.60 mg/mL. |
| Databáze: | OpenAIRE |
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