development and application of aqueous two-phase partition for the recovery and separation of recombinant phenylalanine dehydrogenase
نویسندگان
چکیده
aqueous two-phase systems (atps) have emerged as a powerful extraction method for the downstream processing of bio-molecules. the aim of this work was to investigate the possibility of utilizing atps for the separation of recombinant bacillus sphaericus phenylalanine dehydrogenase (phedh). polyethylene glycol (peg) and ammonium sulfate systems were selected for our experiment. the effect of different elements such as; type and concentration of peg, concentration of (nh4)2so4, ph, phase volume ratio (vr) and tie-line length (tll) on the extraction behavior and selective separation was also studied. desirable conditions for differential partitioning was obtained in 8.5 % (w/w) peg-6000, 17.5 % (w/w) (nh4)2so4 andvr 0.25at ph 8.0. phedh was mainly concentrated into the upper peg-rich phase in all tested systems. the partition coefficient (k), recovery (r %), yield (y %), tll and selectivity were found to be 58.7, 135 %, 94.42 %, 39.89 % (w/w) and 2174, respectively. from the experimental results, it was revealed that the peg molecular weight, (nh4)2so4 concentration, tll and ph of system had strong impacts on partition features. the extraction efficiency was increased with elevation of ph and tll values. in this paper, we described the partitioning behavior in peg/(nh4)2so4 atps in order to evaluate the applicability of atps for partitioning and recovery of phedh.
منابع مشابه
Development and Application of Aqueous Two-Phase Partition for the Recovery and Separation of Recombinant Phenylalanine Dehydrogenase
Aqueous two-phase systems (ATPS) have emerged as a powerful extraction method for the downstream processing of bio-molecules. The aim of this work was to investigate the possibility of utilizing ATPS for the separation of recombinant Bacillus sphaericus phenylalanine dehydrogenase (PheDH). Polyethylene glycol (PEG) and ammonium sulfate systems were selected for our experi...
متن کاملRapid One-Step Separation and Purification of Recombinant Phenylalanine Dehydrogenase in Aqueous Two-Phase Systems
Background: Phenylalanine dehydrogenase (PheDH EC 1.4.1.20) is a NAD+-dependent enzyme that performs the reversible oxidative deamination of L-phenylalanine to phenylpyruvate. It plays an important role in detection and screening of phenylketonuria (PKU) diseases and production of chiral intermediates as well. The main goal of this study was to find a simple and rapid alternative method for pur...
متن کاملrapid one-step separation and purification of recombinant phenylalanine dehydrogenase in aqueous two-phase systems
background: phenylalanine dehydrogenase (phedh ec 1.4.1.20) is a nad+-dependent enzyme that performs the reversible oxidative deamination of l-phenylalanine to phenylpyruvate. it plays an important role in detection and screening of phenylketonuria (pku) diseases and production of chiral intermediates as well. the main goal of this study was to find a simple and rapid alternative method for pur...
متن کاملComparison 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 pyrrolid...
متن کاملRapid one-step separation and purification of recombinant phenylalanine dehydrogenase in aqueous two-phase systems.
BACKGROUND Phenylalanine dehydrogenase (PheDH; EC 1.4.1.20) is a NAD+-dependent enzyme that performs the reversible oxidative deamination of L-phenylalanine to phenylpyruvate. It plays an important role in detection and screening of phenylketonuria (PKU) diseases and production of chiral intermediates as well. The main goal of this study was to find a simple and rapid alternative method for pur...
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عنوان ژورنال:
iranian journal of chemistry and chemical engineering (ijcce)ناشر: iranian institute of research and development in chemical industries (irdci)-acecr
ISSN 1021-9986
دوره 27
شماره 2 2008
میزبانی شده توسط پلتفرم ابری doprax.com
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