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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Bacillus / enzymology*
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Carrier Proteins / metabolism*
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Catalytic Domain
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Molecular Sequence Data
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Oligosaccharides / biosynthesis*
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Prebiotics*
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Protein Engineering / methods*
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Substrate Specificity
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beta-Galactosidase / chemistry
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beta-Galactosidase / genetics
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beta-Galactosidase / metabolism*
Substances
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Carrier Proteins
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Oligosaccharides
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Prebiotics
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beta-Galactosidase
Associated data
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GENBANK/KT153227
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GENBANK/KT153228
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GENBANK/KT153229
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GENBANK/KT153230
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GENBANK/KT153231
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GENBANK/KT153232
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GENBANK/KT153233
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GENBANK/KT153234
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GENBANK/KT153235
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GENBANK/KT153236
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GENBANK/KT153237
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GENBANK/KT153238
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GENBANK/KT153239
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GENBANK/KT153240
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GENBANK/KT153241
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GENBANK/KT153242
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GENBANK/KT153243
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GENBANK/KT153244
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GENBANK/KT153245
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GENBANK/KT153246
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GENBANK/KT153247
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GENBANK/KT153248
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GENBANK/KT153249
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GENBANK/KT153250