Structure-Guided Immobilization of an Evolved Unspecific Peroxygenase
by
Patricia Molina-Espeja 1,†
, Paloma Santos-Moriano 1,2,†, Eva García-Ruiz 1, Antonio Ballesteros 1, Francisco J. Plou 1,* and Miguel Alcalde 1,*
Patricia Molina-Espeja 1,†, Paloma Santos-Moriano 1,2,†, Eva García-Ruiz 1, Antonio Ballesteros 1, Francisco J. Plou 1,* and Miguel Alcalde 1,*
1
Department of Biocatalysis, Institute of Catalysis, CSIC, Cantoblanco, 28049 Madrid, Spain
2
Applied Biotechnology Group, Faculty of Biomedical and Health Sciences, Universidad Europea de Madrid, Urbanización El Bosque, Villaviciosa de Odón, 28670 Madrid, Spain
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Int. J. Mol. Sci. 2019, 20(7), 1627; https://doi.org/10.3390/ijms20071627
Received: 19 February 2019 / Revised: 25 March 2019 / Accepted: 29 March 2019 / Published: 2 April 2019
(This article belongs to the Special Issue Industrial Enzymes: Structure, Function and Applications)
Unspecific peroxygenases (UPOs) are highly promiscuous biocatalyst with self-sufficient mono(per)oxygenase activity. A laboratory-evolved UPO secreted by yeast was covalently immobilized in activated carriers through one-point attachment. In order to maintain the desired orientation without compromising the enzyme’s activity, the S221C mutation was introduced at the surface of the enzyme, enabling a single disulfide bridge to be established between the support and the protein. Fluorescence confocal microscopy demonstrated the homogeneous distribution of the enzyme, regardless of the chemical nature of the carrier. This immobilized biocatalyst was characterized biochemically opening an exciting avenue for research into applied synthetic chemistry.
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Keywords:
directed evolution; structure-guided immobilization; oxyfunctionalization; unspecific peroxygenase
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MDPI and ACS Style
Molina-Espeja, P.; Santos-Moriano, P.; García-Ruiz, E.; Ballesteros, A.; Plou, F.J.; Alcalde, M. Structure-Guided Immobilization of an Evolved Unspecific Peroxygenase. Int. J. Mol. Sci. 2019, 20, 1627.
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