Broadening the Biocatalytic Toolbox-Screening and Expression of New Unspecific Peroxygenases
Unspecific peroxygenases (UPOs) catalyze the selective transfer of single oxygen atoms from peroxides to a broad range of substrates such as un-activated hydrocarbons. Since specific oxyfunctionalizations are among the most-desired reactions in synthetic chemistry, UPOs are of high industrial intere...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Book |
| Published: |
MDPI AG,
2022-01-01T00:00:00Z.
|
| Subjects: | |
| Online Access: | Connect to this object online. |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Summary: | Unspecific peroxygenases (UPOs) catalyze the selective transfer of single oxygen atoms from peroxides to a broad range of substrates such as un-activated hydrocarbons. Since specific oxyfunctionalizations are among the most-desired reactions in synthetic chemistry, UPOs are of high industrial interest. To broaden the number of available enzymes, computational and experimental methods were combined in this study. After a comparative alignment and homology modelling, the enzymes were expressed directly in <i>P. pastoris.</i> Out of ten initially selected sequences, three enzymes (one from <i>Aspergillus niger</i> and two from <i>Candolleomyces aberdarensis</i>) were actively expressed. Cultivation of respective expression clones in a bioreactor led to production titers of up to 300 mg L<sup>−1</sup>. Enzymes were purified to near homogeneity and characterized regarding their specific activities and pH-optima for typical UPO substrates. This work demonstrated that directed evolution is not necessarily required to produce UPOs in <i>P. pastoris</i> at respective titers. The heterologous producibility of these three UPOs will expand the toolbox of available enzymes and help to advance their synthetic application. |
|---|---|
| Item Description: | 10.3390/antiox11020223 2076-3921 |