Functional Expression of Two Unusual Acidic Peroxygenases from <i>Candolleomyces aberdarensis</i> in Yeasts by Adopting Evolved Secretion Mutations
Patricia Gómez de Santos, Manh Dat Hoang, Jan Kiebist, Harald Kellner, René Ullrich, Katrin Scheibner, Martin Hofrichter, Christiane Liers, Miguel Alcalde
Abstract
In this work, we incorporated several secretion mutations from an evolved fungal peroxygenase to enhance the production of active and stable forms of two unusual acidic peroxygenases. The tandem-yeast expression system based on S. cerevisiae for directed evolution and P. pastoris for overproduction on an ∼300-mg/liter scale is a versatile tool to generate UPO variants. By employing this approach, we foresee that acidic UPO variants will be more readily engineered in the near future and adapted to practical enzyme cascade reactions that can be performed over a broad pH range to oxyfunctionalize a variety of organic compounds.