title: Monobody-mediated alteration of enzyme specificity.
abstract: Current methods for engineering enzymes modify enzymes themselves and require a detailed mechanistic understanding or a high-throughput assay. Here, we describe a new approach where catalytic properties are modulated with synthetic binding proteins, termed monobodies, directed to an unmodified enzyme. Using the example of a β-galactosidase from Bacillus circulans, we efficiently identified monobodies that restricted its substrates for its transgalactosylation reaction and selectively enhanced the production of small oligosaccharide prebiotics.
authors: Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois, USA.|Frontier Research Department, Gifu R&D Center, Amano Enzyme, Inc., Gifu, Japan.
authors: Shun-Ichi
authors: S
authors: Tanaka
authors: Frontier Research Department, Gifu R&D Center, Amano Enzyme, Inc., Gifu, Japan.
authors: Tetsuya
authors: T
authors: Takahashi
authors: Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois, USA.
authors: Akiko
authors: A
authors: Koide
authors: Frontier Research Department, Gifu R&D Center, Amano Enzyme, Inc., Gifu, Japan.
authors: Satoru
authors: S
authors: Ishihara
authors: Frontier Research Department, Gifu R&D Center, Amano Enzyme, Inc., Gifu, Japan.
authors: Satoshi
authors: S
authors: Koikeda
authors: Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois, USA.
authors: Shohei
authors: S
authors: Koide
mesh_terms: D001407:Bacillus / Q000201:enzymology*; D002352:Carrier Proteins / Q000378:metabolism*; D020134:Catalytic Domain; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000096:biosynthesis*; D056692:Prebiotics; D015202:Protein Engineering / Q000379:methods*; D013379:Substrate Specificity; D001616:beta-Galactosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D009844:Oligosaccharides; D056692:Prebiotics; D001616:beta-Galactosidase