title: A novel S-enantioselective amidase acting on 3,3,3-trifluoro-2-hydroxy-2-methylpropanamide from Arthrobacter sp. S-2.
abstract: A novel S-enantioselective amidase acting on 3,3,3-trifluoro-2-hydroxy-2-methylpropanamide was purified from Arthrobacter sp. S-2. The enzyme acted S-enantioselectively on 3,3,3-trifluoro-2-hydroxy-2-methylpropanamide to yield (S)-3,3,3-trifluoro-2-hydroxy-2-methylpropanoic acid. Based on the N-terminal amino acid sequence of this amidase, the gene coding S-amidase was cloned from the genomic DNA of Arthrobacter sp. S-2 and expressed in an Escherichia coli host. The recombinant S-amidase was purified and characterized. Furthermore, the purified recombinant S-amidase with the C-His6-tag, which was expressed in E. coli as the C-His6-tag fusion protein, was used in the kinetic resolution of (±)-3,3,3-trifluoro-2-hydroxy-2-methylpropanamide to obtain (S)-3,3,3-trifluoro-2-hydroxy-2-methylpropanoic acid and (R)-3,3,3-trifluoro-2-hydroxy-2-methylpropanamide.
authors.affiliation: a Biotechnology Research Center and Department of Biotechnology , Toyama Prefectural University , Imizu , Japan.
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Fuhshuku
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Watanabe
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Nishii
authors.forename: Akihiro
authors.initials: A
authors.lastname: Ishii
authors.forename: Yasuhisa
authors.initials: Y
authors.lastname: Asano
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000813:Anilides / Q000737:chemistry / Q000378:metabolism*; D001173:Arthrobacter / Q000737:chemistry* / Q000201:enzymology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006639:Histidine / Q000235:genetics / Q000378:metabolism; D006868:Hydrolysis; D006885:Hydroxybutyrates / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D000813:Anilides; D001426:Bacterial Proteins; C471213:His-His-His-His-His-His; D006885:Hydroxybutyrates; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D006639:Histidine; D000581:Amidohydrolases
keywords: 3,3,3-trifluoro-2-hydroxy-2-methylpropanamide; 3,3,3-trifluoro-2-hydroxy-2-methylpropanoic acid; Arthrobacter sp.; amidase