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Production of Recombinant Proline Dehydrogenase Enzyme from Pseudomonas fluorescens pf-5 in E. coli System | ||
| Journal of Sciences, Islamic Republic of Iran | ||
| مقاله 3، دوره 22، شماره 4، اسفند 2011، صفحه 321-327 اصل مقاله (265.65 K) | ||
| نویسنده | ||
| E. Omidinia | ||
| Pasteur Institute of Iran | ||
| چکیده | ||
| 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. | ||
| کلیدواژه‌ها | ||
| Cloning؛ pseudomonas fluorescence؛ Characterization؛ Proline dehydrogenase (ProDH) | ||
| عنوان مقاله [English] | ||
| Production of Recombinant Proline Dehydrogenase Enzyme from Pseudomonas fluorescens pf-5 in E. coli System | ||
| نویسندگان [English] | ||
| E. Omidinia | ||
| چکیده [English] | ||
| 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. | ||
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آمار تعداد مشاهده مقاله: 1,806 تعداد دریافت فایل اصل مقاله: 1,881 |
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