phenylketonuria (pku) is an autosomal recessive disorder, which is characterized by severe mental retardation, microcephaly and seizures. the symptoms of this disease can be prevented if detected soon after birth. therefore, blood phenylalanine (phe) measurement is essential for the early diagnosis, treatment and dietary monitoring of pku patients. the goal of this research was to introduce a r...

We describe here a new microquantification method of l-phenylalanine concentration in an extract from a dried blood spot by using the diaphorase-resazurin system. To miniaturize the fluorometric enzymatic microplate assay for the diagnosis of phenylketonuria, an enzyme chip immobilized with His-tag fused phenylalanine dehydrogenase (PheDH) was developed. His-tag fused PheDH was immobilized on t...

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...

cloning and expression of the l-phenylalanine dehydrogenase gene, from b. sphaericus in e. coli were done. the gene was cloned in the vector pet16b and transformed into e. coli bl21 (de3). the functional form of the l-phenylalanine dehydrogenase enzyme was purified by affinity purification techniques, taking advantage of the ability of this enzyme to bind to the nucleotide site affinity dye, re...

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...

To evaluate the role of wild promoter of L-phenylalanine dehydrogenase (PheDH) gene, referred to as pdh, from Bacillus sphaericus in expression, cloning of pdh gene in Bacillus subtilis was performed. The whole pdh gene was cloned in pHY300PLK shuttle vector and amplified, construct (pHYDH) then transformed in B. subtilis ISW1214 and E. coli JM109. The pdh endogenous promoter presented no effec...

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...

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...

to evaluate the role of wild promoter of l-phenylalanine dehydrogenase (phedh) gene, referred to as pdh, from bacillus sphaericus in expression, cloning of pdh gene in bacillus subtilis was performed. the whole pdh gene was cloned in phy300plk shuttle vector and amplified, construct (phydh) then transformed in b. subtilis isw1214 and e. coli jm109. the pdh endogenous promoter presented no effec...

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 di...