Title
Fungal peroxygenase chimera identified by sub-domain shuffling and split-gfp reveal retained activity and altered specificity
Conference Dates
September 15-19, 2019
Abstract
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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Recommended Citation
Pascal Püllmann and Martin J. Weissenborn, "Fungal peroxygenase chimera identified by sub-domain shuffling and split-gfp reveal retained activity and altered specificity" in "Enzyme Engineering XXV", Huimin Zhao, University of Illinois at Urbana-Champaign, USA John Wong, Pfizer, USA Eds, ECI Symposium Series, (2019). https://dc.engconfintl.org/enzyme_xxv/38