Isolation, Screening and Identification of Bi-functional PutA Flavoenzyme Producing Pseudomonas Fluoresense from Soil

The bifunctional proline utilization A (PutA) contains proline dehydrogenase and ∆-pyrroline-5-carboxylate dehydrogenase domains and plays an important role in the metabolic pathway of proline to glutamate. The aim of this research was to isolate and identify the PutA falvoenzyme producing bacteria with appropriate kinetic properties. Needed soil samples were collected from different places of Tehran city including Darakeh, Darband, Farahzad and Damavand. Isolation and screening of falvoenzyme producing microorganisms was performed by enrichment culture technique in proline specific culture medium and enzyme activity staining by formazan color, reaction and confirmed by enzyme assay. Isolated strain was identified by phenotypic and biochemical characteristics. specific activity of falvoenzyme was also measured. In total of 400 isolated bacteria, only one of them was PutA producer that recognized as Pseudomonas fluorescens. The specific activity of falvoenzyme for L-proline and ∆1-Pyrroline-5-carboxylate was determined 22.4 U/mg and 10.7 U/mg, respectively. Specific activity of this enzyme for proline and ∆-pyrroline-5-carboxylate in compression with various amount of falvoenzyme which isolated from other resources is acceptable.