Production of Recombinant Proline Dehydrogenase Enzyme from Pseudomonas fluorescens pf-5 in E. coli System
Proline dehydrogenase (ProDH; 1.5.99.8) belongs to superfamily of amino acid dehydrogenase, which plays a significant role in the metabolic pathway from proline to glutamate. The goal of this research was gene cloning and characterization of ProDH enzyme from Pseudomonas fluorescens pf-5 strain. The...
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University of Tehran
2011-12-01
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| Series: | Journal of Sciences, Islamic Republic of Iran |
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| Online Access: | https://jsciences.ut.ac.ir/article_23866_63a6e5e7500846f1c48213192b466b42.pdf |
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| author | E. Omidinia |
| author_facet | E. Omidinia |
| author_sort | E. Omidinia |
| collection | DOAJ |
| description | Proline dehydrogenase (ProDH; 1.5.99.8) belongs to superfamily of amino acid dehydrogenase, which plays a significant role in the metabolic pathway from proline to glutamate. The goal of this research was gene cloning and characterization of ProDH enzyme from Pseudomonas fluorescens pf-5 strain. The gene encoding ProDH was isolated by means of PCR amplification and cloned in an IPTG inducible T7-based expression system. The Histidine-tagged recombinant enzyme was purified and its kinetic properties were studied. According to SDS-PAGE analysis ProDH revealed a MW of 40 kDa. The Km and Vmax values of P. fluorescens ProDH were estimated to be 20 mM and 160 ?mol/min, respectively. ProDH activity was stable at alkaline pH and the highest activity was observed at pH 8.5 and 30°C. This study is the first data on the isolation and production of P. fluorescens ProDH enzyme in E. coli expression system. |
| format | Article |
| id | doaj-art-0b15fee2f06744af9699fcd0988ad4ae |
| institution | Kabale University |
| issn | 1016-1104 2345-6914 |
| language | English |
| publishDate | 2011-12-01 |
| publisher | University of Tehran |
| record_format | Article |
| series | Journal of Sciences, Islamic Republic of Iran |
| spelling | doaj-art-0b15fee2f06744af9699fcd0988ad4ae2025-08-20T03:53:51ZengUniversity of TehranJournal of Sciences, Islamic Republic of Iran1016-11042345-69142011-12-0122432132723866Production of Recombinant Proline Dehydrogenase Enzyme from Pseudomonas fluorescens pf-5 in E. coli SystemE. Omidinia0Pasteur Institute of IranProline dehydrogenase (ProDH; 1.5.99.8) belongs to superfamily of amino acid dehydrogenase, which plays a significant role in the metabolic pathway from proline to glutamate. The goal of this research was gene cloning and characterization of ProDH enzyme from Pseudomonas fluorescens pf-5 strain. The gene encoding ProDH was isolated by means of PCR amplification and cloned in an IPTG inducible T7-based expression system. The Histidine-tagged recombinant enzyme was purified and its kinetic properties were studied. According to SDS-PAGE analysis ProDH revealed a MW of 40 kDa. The Km and Vmax values of P. fluorescens ProDH were estimated to be 20 mM and 160 ?mol/min, respectively. ProDH activity was stable at alkaline pH and the highest activity was observed at pH 8.5 and 30°C. This study is the first data on the isolation and production of P. fluorescens ProDH enzyme in E. coli expression system.https://jsciences.ut.ac.ir/article_23866_63a6e5e7500846f1c48213192b466b42.pdfcloningpseudomonas fluorescencecharacterizationproline dehydrogenase (prodh) |
| spellingShingle | E. Omidinia Production of Recombinant Proline Dehydrogenase Enzyme from Pseudomonas fluorescens pf-5 in E. coli System Journal of Sciences, Islamic Republic of Iran cloning pseudomonas fluorescence characterization proline dehydrogenase (prodh) |
| title | Production of Recombinant Proline Dehydrogenase Enzyme from Pseudomonas fluorescens
pf-5 in E. coli System |
| title_full | Production of Recombinant Proline Dehydrogenase Enzyme from Pseudomonas fluorescens
pf-5 in E. coli System |
| title_fullStr | Production of Recombinant Proline Dehydrogenase Enzyme from Pseudomonas fluorescens
pf-5 in E. coli System |
| title_full_unstemmed | Production of Recombinant Proline Dehydrogenase Enzyme from Pseudomonas fluorescens
pf-5 in E. coli System |
| title_short | Production of Recombinant Proline Dehydrogenase Enzyme from Pseudomonas fluorescens
pf-5 in E. coli System |
| title_sort | production of recombinant proline dehydrogenase enzyme from pseudomonas fluorescens pf 5 in e coli system |
| topic | cloning pseudomonas fluorescence characterization proline dehydrogenase (prodh) |
| url | https://jsciences.ut.ac.ir/article_23866_63a6e5e7500846f1c48213192b466b42.pdf |
| work_keys_str_mv | AT eomidinia productionofrecombinantprolinedehydrogenaseenzymefrompseudomonasfluorescenspf5inecolisystem |