Directed evolution of a Swiss knife ligninase: The unspecific peroxygenase
September 24-28, 2017
Among the broad repertory of the ligninolytic enzyme consortium, the unspecific peroxygenase (UPO) is shining as a versatile biocatalyst with potential applications in organic chemistry. UPO is a heme-thiolate peroxidase with mono(per)oxygenase activity for the selective oxyfunctionalization of C-H bonds. Fueled by catalytic concentrations of H2O2, which acts as both oxygen donor and as final electron acceptor, this stable, soluble and extracellular enzyme performs dozens of transformations that are of considerable interest in organic synthesis. In this talk I will describe the main attributes of this versatile enzyme, while reflecting on the directed evolution campaigns recently followed in our laboratory that set out to enhance the functional expression of UPO in yeast and improve the activity, as well as approximating its properties to the required industrial standards.
Mate, D.M., Palomino, M.A., Molina-Espeja, P., Martin-Diaz, J. and Alcalde, M. (2017). Modification of the peroxygenative:peroxidative activity ratio in the unspecific peroxygenase from Agrocybe aegerita by structure-guided evolution. Protein Engineering Design and Selection. In press.
Molina-Espeja, P., Gomez de Santos, P., and Alcalde, M. (2017). Directed evolution of unspecific peroxygenase. In: Directed Enzyme Evolution: Advances and Applications. Alcalde, M. (Ed). Springer. 127-143.
Molina-Espeja, P., Cañellas, M., Plou, F.J., Hofrichter, M., Lucas, F., Guallar, V. and Alcalde, M. (2016). Synthesis of 1-naphthol by a natural peroxygenase engineered by directed evolution. ChemBioChem 17: 341-349.
Molina-Espeja, P., Ma, S., Mate, D.M., Ludwig, R. and Alcalde, M. (2015). Tandem-yeast expression system for engineering and producing unspecific peroxygenase. Enzyme Microbial Technology 73-74:29-33
Alcalde, M. (2015). Engineering the ligninolytic enzyme consortium. Trends in Biotechnology 33: 155-162.
Molina-Espeja, P., Garcia-Ruiz, E., Gonzalez-Perez, D., Ullrich, R., Hofrichter, M. and Alcalde, M. (2014). Directed evolution of unspecific peroxygenase from Agrocybe aegerita. Applied & Environmental Microbiology 80: 3496-3507
Miguel Alcalde, "Directed evolution of a Swiss knife ligninase: The unspecific peroxygenase" in "Enzyme Engineering XXIV", Pierre Monsan, Toulouse White Biotechnology, France Magali Remaud-Simeon, LISBP-INSA, University of Toulouse, France Eds, ECI Symposium Series, (2017). http://dc.engconfintl.org/enzyme_xxiv/119