Production of Recombinant Proline Dehydrogenase Enzyme from Pseudomonas fluorescens pf-5 in E. coli System
نویسنده
چکیده مقاله:
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.
منابع مشابه
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 gene encoding prodh was isolated by means of pcr amplification and cloned in an iptg inducible t...
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عنوان ژورنال
دوره 22 شماره 4
صفحات 321- 327
تاریخ انتشار 2011-12-01
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