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Pilot-Scale Production of Peroxygenase from Agrocybe aegerita.

Fabio ToninFlorian TievesSébastien WillotAnouska van TroostRemco van OostenStefaan BreestraatSander van PeltMiguel AlcaldeFrank Hollmann
Published in: Organic process research & development (2021)
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).
Keyphrases
  • study protocol
  • saccharomyces cerevisiae
  • lactic acid
  • randomized controlled trial
  • gram negative
  • mass spectrometry
  • ionic liquid
  • high resolution
  • anaerobic digestion
  • liquid chromatography
  • tandem mass spectrometry