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Title

Synthesis of 1‐Naphthol by a Natural Peroxygenase Engineered by Directed Evolution

AuthorsMolina-Espeja, Patricia CSIC ORCID ; Cañelas, Marina; Plou Gasca, Francisco José CSIC ORCID ; Hofrichter, Martin; Lucas, Fátima; Guallar, Victor; Alcalde Galeote, Miguel CSIC ORCID
Keywords1-naphthol
Directed evolution
Enzyme catalysis
Peroxidative activity
Peroxygenases
Issue Date2015
PublisherJohn Wiley & Sons
CitationChemBioChem 17: 341-349 (2016)
AbstractThere is an increasing interest in enzymes that catalyze the hydroxylation of naphthalene under mild conditions and with minimal requirements. To address this challenge, an extracellular fungal aromatic peroxygenase with mono(per)oxygenase activity was engineered to convert naphthalene selectively into 1‐naphthol. Mutant libraries constructed by random mutagenesis and DNA recombination were screened for peroxygenase activity on naphthalene together with quenching of the undesired peroxidative activity on 1‐naphthol (one‐electron oxidation). The resulting double mutant (G241D‐R257K) obtained from this process was characterized biochemically and computationally. The conformational changes produced by directed evolution improved the substrate's catalytic position. Powered exclusively by catalytic concentrations of H2O2, this soluble and stable biocatalyst has a total turnover number of 50 000, with high regioselectivity (97 %) and reduced peroxidative activity.
Publisher version (URL)https://doi.org/10.1002/cbic.201500493
URIhttp://hdl.handle.net/10261/171077
DOI10.1002/cbic.201500493
ISSN1439-4227
E-ISSN1439-7633
Appears in Collections:(ICP) Artículos

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