title: Purification and characterization of enantioselective N-acetyl-β-Phe acylases from Burkholderia sp. AJ110349.
abstract: For the production of enantiopure β-amino acids, enantioselective resolution of N-acyl β-amino acids using acylases, especially those recognizing N-acetyl-β-amino acids, is one of the most attractive methods. Burkholderia sp. AJ110349 had been reported to exhibit either (R)- or (S)-enantiomer selective N-acetyl-β-Phe amidohydrolyzing activity, and in this study, both (R)- and (S)-enantioselective N-acetyl-β-Phe acylases were purified to be electrophoretically pure and determined the sequences, respectively. They were quite different in terms of enantioselectivities and in their amino acids sequences and molecular weights. Although both the purified acylases were confirmed to catalyze N-acetyl hydrolyzing activities, neither of them show sequence similarities to the N-acetyl-α-amino acid acylases reported thus far. Both (R)- and (S)-enantioselective N-acetyl-β-Phe acylase were expressed in Escherichia coli. Using these recombinant strains, enantiomerically pure (R)-β-Phe (>99% ee) and (S)-β-Phe (>99% ee) were obtained from the racemic substrate.
authors.affiliation: a AminoScience Laboratories , Ajinomoto Co., Ltd. , Kawasaki , Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Imabayashi
authors.affiliation: a AminoScience Laboratories , Ajinomoto Co., Ltd. , Kawasaki , Japan.
authors.forename: Shun'ichi
authors.initials: S
authors.lastname: Suzuki
authors.affiliation: b Department of Environmental Materials Science , Tokyo Denki University , Adachi-ku , Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Kawasaki
authors.affiliation: b Department of Environmental Materials Science , Tokyo Denki University , Adachi-ku , Japan.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Nakamatsu
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D055162:Biocatalysis; D019117:Burkholderia / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D057075:Enzyme Assays; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006868:Hydrolysis; D007700:Kinetics; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000581:Amidohydrolases; C019665:amidase
keywords: Burkholderia; N-acetyl-β-amino acid; enantiomer selective amidohydrolyzing activity; β-amino acid