title: C-Terminal-oriented Immobilization of Enzymes Using Sortase A-mediated Technique.
abstract: In the present study, sortase A-mediated immobilization of enzymes was used for the preparation of immobilized enzymes. Thermobifida fusca YX β-glucosidase (BGL) or Streptococcus bovis 148 α-amylase (AmyA) were produced with C-terminal sortase A recognition sequences. The resulting fusion proteins were successfully immobilized on nanoparticle surfaces using sortase A. Some properties (activity, stability, and reusability) of the immobilized fusion proteins were evaluated. Both immobilized BGL and immobilized AmyA prepared by the sortase A-mediated technique retained their catalytic activity, exhibiting activities 3.0- or 1.5-fold (respectively) of those seen with the same enzymes immobilized by chemical crosslinking. Immobilized enzymes prepared by the sortase A-mediated technique did not undergo dramatic changes in stability compared with the respective free enzymes. Thus, the sortase A-mediated technique provides a promising method for immobilization of active, stable enzymes.
authors.affiliation: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
authors.forename: Yuto
authors.initials: Y
authors.lastname: Hata
authors.affiliation: Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Matsumoto
authors.affiliation: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan. tanaka@kitty.kobe-u.ac.jp.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kondo
mesh_terms: D019881:Aminoacyltransferases / Q000737:chemistry* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D001671:Biochemistry / Q000379:methods*; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000378:metabolism*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D053758:Nanoparticles / Q000737:chemistry; D059027:Recycling; D013696:Temperature; D000516:alpha-Amylases / Q000378:metabolism*; D001617:beta-Glucosidase / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D019881:Aminoacyltransferases; C120542:sortase A; D000516:alpha-Amylases; D001617:beta-Glucosidase; D003546:Cysteine Endopeptidases
keywords: enzymes; immobilization of enzymes; sortase A