Pilot-scale production of peroxygenase from agrocybe aegerita

Pilot-scale production of peroxygenase from agrocybe aegerita

Tonin, F. (TU Delft BT/Biocatalysis)
Tieves, F. (TU Delft BT/Biocatalysis)
Willot, S.J. (TU Delft BT/Biocatalysis)
Van Troost, Anouska (Student TU Delft)
van Oosten, R.J.C. (TU Delft BT/Biocatalysis)
Breestraat, Stefaan (Bioprocess Pilot Facility)
Van Pelt, Sander (Bioprocess Pilot Facility)
Alcalde, Miguel (Institute of Catalysis, CSIC, Madrid)
Hollmann, F. (TU Delft BT/Biocatalysis)

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).

Biocatalysis
Hydroxylation
Peroxygenases
Pilot-scale fermentation
Specific oxyfunctionalization

http://resolver.tudelft.nl/uuid:dff69288-2057-4fa5-a71d-290f4b1553d0

https://doi.org/10.1021/acs.oprd.1c00116

1083-6160

Organic Process Research and Development, 25 (6), 1414-1418

Institutional Repository

journal article

© 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