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 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