Pilot-scale production of peroxygenase from agrocybe aegerita

Journal Article (2021)
Authors

Fabio Tonin (TU Delft - BT/Biocatalysis)

F. Tieves (TU Delft - BT/Biocatalysis)

Sébastien Jean Paul Willot (TU Delft - BT/Biocatalysis)

Anouska Van Troost (Student TU Delft)

Remco Van Oosten (TU Delft - BT/Biocatalysis)

Stefaan Breestraat (Bioprocess Pilot Facility)

Sander Van Pelt (Bioprocess Pilot Facility)

Miguel Alcalde (Institute of Catalysis, CSIC, Madrid)

Frank Hollmann (TU Delft - BT/Biocatalysis)

Research Group
BT/Biocatalysis
Copyright
© 2021 F. Tonin, F. Tieves, S.J. Willot, Anouska Van Troost, R.J.C. van Oosten, Stefaan Breestraat, Sander Van Pelt, Miguel Alcalde, F. Hollmann
To reference this document use:
https://doi.org/10.1021/acs.oprd.1c00116
More Info
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Publication Year
2021
Language
English
Copyright
© 2021 F. Tonin, F. Tieves, S.J. Willot, Anouska Van Troost, R.J.C. van Oosten, Stefaan Breestraat, Sander Van Pelt, Miguel Alcalde, F. Hollmann
Research Group
BT/Biocatalysis
Issue number
6
Volume number
25
Pages (from-to)
1414-1418
DOI:
https://doi.org/10.1021/acs.oprd.1c00116
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Abstract

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).