comparison between polyvinyl pyrrolidone/na2so4 aqueous two-phase systems and chromatographic methods for purification of recombinant phenylalanine dehydrogenase

phenylalanine dehydrogenase (phedh; ec 1.4.1.20) is an important enzyme of amino acid dehydrogenases family that increasingly used as a valuable biocatalyst in neonatal screening kits and synthesis of l-phenylalanine. the goal of this literature was to find a suitable purification method for recombinant bacillus badius phedh by practical comparison between chromatographic and polyvinyl pyrrolidone (pvp)/na2so4 aqueous two-phase systems (atps) techniques. the partitioning behavior of target enzyme in pvp/na2so4 atps was examined and compared with the obtained results from a chromatographic protocol. direct comparison of chromatography and atps procedures clearly revealed that the atps consisting of 8.0% (w/w) pvp, 17.0% (w/w) na2so4 with ph of 8.0, vr=0.25 and temperature of 25 °c was the most desirable process for phedh purification. a specific activity of 1231.42 u/mg, a purification factor of 36.61, a yield of 95.5% and a recovery of 138.9% were achieved. altogether, we presented a two-phase methodology as a scalable and economically alternative for the production of phedh enzyme.