Discovery and heterologous expression of unspecific Peroxygenases

Katharina Ebner, Lukas Pfeifenberger, Veronika Schusterbauer, Claudia Rinnofner, Anton Glieder*, Margit Winkler*

*Korrespondierende/r Autor/-in für diese Arbeit

Publikation: Beitrag in einer FachzeitschriftArtikelBegutachtung

Abstract

Since 2004, unspecific peroxygenases, in short UPOs (EC. 1.11.2.1), have been explored. UPOs are closing a gap between P450 monooxygenases and chloroperoxidases. These enzymes are highly active biocatalysts for the selective oxyfunctionalisation of C–H, C=C and C-C bonds. UPOs are secreted fungal proteins and Komagataella phaffii (Pichia pastoris) is an ideal host for high throughput screening approaches and UPO production. Heterologous overexpression of 26 new UPOs by K. phaffii was performed in deep well plate cultivation and shake flask cultivation 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 and 5-nitro-1,3-benzodioxole (NBD) as reporter substrates. The PaDa-I (AaeUPO mutant) and HspUPO were used as benchmarks to find interesting new enzymes with complementary activity profiles as well as good producing strains. Herein we show that six UPOs from Psathyrella aberdarensis, Coprinopsis marcescibilis, Aspergillus novoparasiticus, Dendrothele bispora and Aspergillus brasiliensis are particularly active.
Originalspracheenglisch
Aufsatznummer206
FachzeitschriftCatalysts
Jahrgang13
Ausgabenummer1
DOIs
PublikationsstatusVeröffentlicht - 6 Jan. 2023

ASJC Scopus subject areas

  • Umweltwissenschaften (insg.)
  • Katalyse
  • Physikalische und Theoretische Chemie

Fields of Expertise

  • Human- & Biotechnology

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