International journal of molecular sciences, 2020-10, Vol.21 (21), p.7859
2020
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- Autor(en) / Beteiligte
- Titel
- Phe-140 Determines the Catalytic Efficiency of Arylacetonitrilase from Alcaligenes faecalis
- Ist Teil von
- International journal of molecular sciences, 2020-10, Vol.21 (21), p.7859
- Ort / Verlag
- Switzerland: MDPI AG
- Erscheinungsjahr
- 2020
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- Arylacetonitrilase from ATCC8750 (NitAF) hydrolyzes various arylacetonitriles to the corresponding carboxylic acids. A systematic strategy of amino acid residue screening through sequence alignment, followed by homology modeling and biochemical confirmation was employed to elucidate the determinant of NitAF catalytic efficiency. Substituting Phe-140 in NitAF (wild-type) to Trp did not change the catalytic efficiency toward phenylacetonitrile, an arylacetonitrile. The mutants with nonpolar aliphatic amino acids (Ala, Gly, Leu, or Val) at location 140 had lower activity, and those with charged amino acids (Asp, Glu, or Arg) exhibited nearly no activity for phenylacetonitrile. Molecular modeling showed that the hydrophobic benzene ring at position 140 supports a mechanism in which the thiol group of Cys-163 carries out a nucleophilic attack on a cyanocarbon of the substrate. Characterization of the role of the Phe-140 residue demonstrated the molecular determinant for the efficient formation of arylcarboxylic acids.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1422-0067, 1661-6596eISSN: 1422-0067DOI: 10.3390/ijms21217859Titel-ID: cdi_doaj_primary_oai_doaj_org_article_a63d94f0633844b9ae377332b21aac2d
- Format
- –
- Schlagworte
- Acetonitriles - metabolism, Alcaligenes faecalis, Alcaligenes faecalis - enzymology, Alcaligenes faecalis - genetics, Amino Acid Sequence, Amino Acid Substitution, Amino acids, Aminohydrolases - chemistry, Aminohydrolases - genetics, Aminohydrolases - metabolism, Arylacetonitrilase, Bacterial Proteins - chemistry, Bacterial Proteins - genetics, Bacterial Proteins - metabolism, Benzene, Carboxylic acids, Catalytic Domain, catalytic efficiency, Efficiency, Enzyme kinetics, Homology, Hydrophobic and Hydrophilic Interactions, Hydrophobicity, Modelling, Models, Molecular, Molecular modelling, Mutagenesis, Mutation, Nucleotide sequence, Phenylalanine - metabolism, Protein Conformation, Proteins, Residues, Simulation, Substrates