title: Amino acid alterations essential for increasing the catalytic activity of the nylon-oligomer-degradation enzyme of Flavobacterium sp.
abstract: The structural genes of two homologous enzymes, 6-aminohexanoate-dimer hydrolase (EII; nylB) and its evolutionally related protein EII' (nylB') of Flavobacterium sp. KI72 have an open reading frame encoding a peptide of 392 amino acids, of which 47 are different, and conserved restriction sites. The specific activity of EII towards 6-aminohexanoate dimer is about 1000-fold that of EII'. Construction of various hybrid genes obtained by exchanging fragments flanked by conserved restriction sites of the two genes demonstrated that two amino acid replacements in the EII' enzyme, i.e. Gly181----Asp (EII type) and His266----Asn (EII type), enhanced the activity toward 6-aminohexanoate dimer 1000-fold.
authors.affiliation: Department of Fermentation Technology, Osaka University, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Kato
authors.forename: K
authors.initials: K
authors.lastname: Fujiyama
authors.forename: H S
authors.initials: HS
authors.lastname: Hatanaka
authors.forename: I D
authors.initials: ID
authors.lastname: Priyambada
authors.forename: S
authors.initials: S
authors.lastname: Negoro
authors.forename: I
authors.initials: I
authors.lastname: Urabe
authors.forename: H
authors.initials: H
authors.lastname: Okada
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism*; D001483:Base Sequence; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D005417:Flavobacterium / Q000201:enzymology*; D008969:Molecular Sequence Data; D009154:Mutation; D009841:Oligonucleotides; D016366:Open Reading Frames; D010957:Plasmids; D012689:Sequence Homology, Nucleic Acid; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D009841:Oligonucleotides; D000581:Amidohydrolases; C026250:6-aminohexanoate-dimer hydrolase; C019683:6-aminohexanoate-cyclic-dimer hydrolase