title: Construction of a tailor-made L (2S,3S)-butanediol dehydrogenase by exchanging domains between native structural analogs.
abstract: The development of a stable L-BDH chimera was attempted by exchanging whole domains between two native structural analogs, L-BDH and meso-BDH, because the S-configuration specificity of L-BDH is valuable from the standpoint of its application but its activity is unstable, whereas meso-BDH is stable. The domain chimeras obtained indicated that the leaf-like structures constituting three domains were likely to be mainly associated with chiral recognition, and the fourth domain, the basic domain, is likely to be mainly associated with enzyme stability. A combination of the leaf domains of L-BDH and the basic domain of meso-BDH attained a sufficient level of practical use as an artificial L-BDH chimera, because the resulting enzyme had both stability and S-configuration specificity. However, the levels of stability and specificity were slightly lower than those of the respective enzymes from which they were derived.
authors: Graduate School of Medicine and Engineering, University of Yamanashi, Takeda, Kofu, Japan.
authors: Tomohito
authors: T
authors: Shimegi
authors: Yuhsuke
authors: Y
authors: Takusagawa
authors: Takashi
authors: T
authors: Ohtsuki
authors: Satoko
authors: S
authors: Noda
authors: Genji
authors: G
authors: Kurisu
authors: Masami
authors: M
authors: Kusunoki
authors: Sadaharu
authors: S
authors: Ui
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D000429:Alcohol Oxidoreductases; C019801:butanediol dehydrogenase