Discovery and Heterologous Expression of Unspecific Peroxygenases

Since 2004, unspecific peroxygenases, in short UPOs (EC. 1.11.2.1), have been explored. are closing a gap between P450 monooxygenases and chloroperoxidases. These enzymes highly active biocatalysts for the selective oxyfunctionalisation of C–H, C=C C-C bonds. secreted fungal proteins Komagataella phaffii (Pichia pastoris) is an ideal host high throughput screening approaches UPO production. Heterologous overexpression 26 new by K. was performed deep well plate cultivation shake flask up to 50 mL volume. Enzymes were screened using colorimetric assays with 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,6-dimethoxyphenol (DMP), naphthalene 5-nitro-1,3-benzodioxole (NBD) as reporter substrates. The PaDa-I (AaeUPO mutant) HspUPO used benchmarks find interesting complementary activity profiles good producing strains. Herein we show that six from Psathyrella aberdarensis, Coprinopsis marcescibilis, Aspergillus novoparasiticus, Dendrothele bispora brasiliensis particularly active.