title: Deduced catalytic mechanism of D-amino acid amidase from Ochrobactrum anthropi SV3.
abstract: D-Amino acid amidase (DAA) from Ochrobactrum anthropi SV3 catalyzes D-stereospecific hydrolysis of amino acid amides. DAA has attracted attention as a catalyst for the stereospecific production of D-amino acids, although the mechanism that drives the reaction has not been clear. Previously, the structure of DAA was classified into two types, a substrate-bound state with an ordered Omega loop, and a ground state with a disordered Omega loop. Because the binding of the substrate facilitates ordering, this transition was regarded to be induced fit motion. The angles and distances of hydrogen bonds at Tyr149 Oeta, Ser60 Ogamma and Lys63 Nzeta revealed that Tyr149 Oeta donates an H atom to a water molecule in the substrate-bound state, and that Tyr149 Oeta donates an H atom to Ser60 Ogamma or Lys63 Nzeta in the ground state. Taking into consideration the locations of the H atoms of Tyr149 Oeta, Ser60 Ogamma and Lys63 Nzeta, a catalytic mechanism of DAA activity is presented, wherein a shift of an H atom at Tyr149 Oeta in the substrate-bound versus the ground state plays a significant role in the reaction. This mechanism explains well why acylation proceeds and deacylation does not proceed in the substrate-bound state.
authors.affiliation: Department of Biotechnology, School of Engineering, Nagoya University, Chikusa, Nagoya 464-8603, Japan.
authors.forename: Seiji
authors.initials: S
authors.lastname: Okazaki
authors.forename: Atsuo
authors.initials: A
authors.lastname: Suzuki
authors.forename: Hidenobu
authors.initials: H
authors.lastname: Komeda
authors.forename: Yasuhisa
authors.initials: Y
authors.lastname: Asano
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamane
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry / Q000378:metabolism*; D002384:Catalysis; D006860:Hydrogen Bonding; D008958:Models, Molecular; D020621:Ochrobactrum anthropi / Q000201:enzymology*
chemical_list: D000581:Amidohydrolases; C407355:D-amino acid amidase