Synthesis of 1‐Naphthol by a Natural Peroxygenase engineered by Directed Evolution
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Projecte de la Comissió EuropeaOptimized oxidoreductases for medium and large scale industrial biotransformations (EC-FP7-613549)
There 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.
This is the peer reviewed version of the following article, which has been published in final form at 10.1002/cbic.201500493. This article may be used for non-commercial purposes in accordance With Wiley-VCH Terms and Conditions for self-archiving
CitacióMolina-Espeja, Patricia [et al.]. Synthesis of 1‐Naphthol by a Natural Peroxygenase engineered by Directed Evolution. "ChemBioChem", 21 Gener 2016, vol. 17, núm. 4, p. 341-349.
Versió de l'editorhttp://onlinelibrary.wiley.com/doi/10.1002/cbic.201500493/abstract