Pilot-scale production of peroxygenase from agrocybe aegerita

Fabio Tonin*, Florian Tieves, Sébastien Willot, Anouska Van Troost, Remco Van Oosten, Stefaan Breestraat, Sander Van Pelt, Miguel Alcalde, Frank Hollmann

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)
30 Downloads (Pure)


The pilot-scale production of the peroxygenase from Agrocybe aegerita (rAaeUPO) is demonstrated. In a fed-batch fermentation of the recombinant Pichia pastoris, the enzyme was secreted into the culture medium to a final concentration of 0.29 g L-1 corresponding to 735 g of the peroxygenase in 2500 L of the fermentation broth after 6 days. Due to nonoptimized downstream processing, only 170 g of the enzyme has been isolated. The preparative usefulness of the so-obtained enzyme preparation has been demonstrated at a semipreparative scale (100 mL) as an example of the stereoselective hydroxylation of ethyl benzene. Using an adjusted H2O2 feed rate, linear product formation was observed for 7 days, producing more than 5 g L-1 (R)-1-phenyl ethanol. The biocatalyst performed more than 340.000 catalytic turnovers (942 g of the product per gram of rAaeUPO).

Original languageEnglish
Pages (from-to)1414-1418
Number of pages5
JournalOrganic Process Research and Development
Issue number6
Publication statusPublished - 2021


  • Biocatalysis
  • Hydroxylation
  • Peroxygenases
  • Pilot-scale fermentation
  • Specific oxyfunctionalization


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