Single Amino Acid Substitution in Bacillus sphaericus Phenylalanine Dehydrogenase Dramatically Increases Its Discrimination between Phenylalanine and Tyrosine Substrates
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منابع مشابه
Single amino acid substitution in Bacillus sphaericus phenylalanine dehydrogenase dramatically increases its discrimination between phenylalanine and tyrosine substrates.
Homology-based modeling of phenylalanine dehydrogenases (PheDHs) from various sources, using the structures of homologous enzymes Clostridium symbiosum glutamate dehydrogenase and Bacillus sphaericus leucine dehydrogenase as a guide, revealed that an asparagine residue at position 145 of B. sphaericus PheDH was replaced by valine or alanine in PheDHs from other sources. This difference was prop...
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Replacement of glycine by serine at positions 123 and 124 of phenylalanine dehydrogenases from Bacillus badius and Bacillus sphaericus respectively strikingly decreased enzyme activity toward aromatic amino acids and resulted in an elevation of relative activity toward aliphatic amino acids. The mutant from B. badius preferentially dehydrogenated branched-chain amino acids, while that from B. s...
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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...
متن کاملAffinity Purification and Characterization of Recombinant Bacillus sphaericus Phenylalanine Dehydrogenase Produced by pET Expression Vector System
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...
متن کاملmolecular cloning and evaluation of wild promoter in expression of bacillus sphaericus phenylalanine dehydrogenase gene in bacillus subtilis cells
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...
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ژورنال
عنوان ژورنال: Biochemistry
سال: 2002
ISSN: 0006-2960,1520-4995
DOI: 10.1021/bi020196a