title: Phenylalanine dehydrogenase of Bacillus badius. Purification, characterization and gene cloning.
abstract: Phenylalanine dehydrogenase produced by Bacillus badius IAM 11059 was purified from the crude extract of B. badius to homogeneity, as judged by disc gel electrophoresis. The enzyme has an isoelectric point of 3.5 and a relative molecular mass, Mr, of 310,000-360,000. The enzyme is composed of identical subunits with an Mr 41,000-42,000. The substrate specificity of the enzyme in the oxidative deamination reaction was high for L-phenylalanine, but rather low in the reductive amination reaction, with phenylpyruvate, p-hydroxyphenylpyruvate, and 2-oxohexanoate. The gene for the enzyme was cloned into Escherichia coli with plasmid pBR322 as a vector. The enzyme was expressed in high level in E. coli. The enzyme produced by E. coli transformant was purified to homogeneity and shown to be identical to that of B. badius IAM 11,059 with respect to the specific activity, Mr, subunit structure and amino acid composition.
authors.affiliation: Sagami Chemical Research Center, Kanagawa, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Asano
authors.forename: A
authors.initials: A
authors.lastname: Nakazawa
authors.forename: K
authors.initials: K
authors.lastname: Endo
authors.forename: Y
authors.initials: Y
authors.lastname: Hibino
authors.forename: M
authors.initials: M
authors.lastname: Ohmori
authors.forename: N
authors.initials: N
authors.lastname: Numao
authors.forename: K
authors.initials: K
authors.lastname: Kondo
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000235:genetics / Q000302:isolation & purification*; D000596:Amino Acids / Q000302:isolation & purification; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D005075:Biological Evolution; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005822:Genetic Vectors; D007526:Isoelectric Point; D010957:Plasmids; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D004269:DNA, Bacterial; D000594:Amino Acid Oxidoreductases; C022342:phenylalanine oxidase