Fungal peroxygenase chimera identified by sub-domain shuffling and split-gfp reveal retained activity and altered specificity

Conference Dates

September 15-19, 2019


Fungal peroxygenases[1] catalyse the hydroxylation of aromatic and aliphatic C-H bonds with high activities, selectivities and thereby relying solely on hydrogen peroxide as a cofactor.[2] Although there are more than 4000 putative fungal peroxygenase genes annotated, only Agrocybe aegerita unspecific peroxygenase (AaeUPO) could be heterologously expressed in S. cerevisiae after five rounds of directed evolution and screening more than 9000 transformants.[3] Four of the identified nine mutations were located in the signal-peptide. However, as “you get what you screen for” in directed evolution, the enzyme has been evolved towards the substrate and not just on the increased expression level. Many fungal genomes carry several dozens of UPO genes suggesting they harbour different specificities and perhaps even catalytic activities. This requires a substrate unspecific screening system for the evolution of these UPOs for heterologous expression.

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