Teetz, NiklasNiklasTeetzLang, SelinaSelinaLangLiese, AndreasAndreasLieseHoltmann, DirkDirkHoltmann2024-11-282024-11-282024-11-11ChemCatChem 16 (21): e202400908 (2024-11-11)https://hdl.handle.net/11420/52180Unspecific peroxygenases (UPOs) are regarded as a “dream catalyst” for selective oxyfunctionalization reactions like oxygenations. We present the display of the model UPO rAaeUPO (PaDa−I) on the cell surface of the heterologous production host Komagataella phaffii as a one-step production and immobilization process. The coding sequence for PaDa−I was combined with genes coding for cell wall proteins from Saccharomyces cerevisiae and transformed into K. phaffii. The fusion proteins were compared among each other and with secreted, free PaDa−I. One system in particular, a C-terminal fusion of PaDa−I and Sag1 yielded near identical activity per volume culture broth to the secreted PaDa−I with ~90 % of the activity being at the cell wall. The surface display simplifies downstream processing and includes immobilization on a cheap, retainable and replaceable matrix, that is the production host itself. The enzymes remained active in a repeated batch process for 10 batches and 200 h of catalysis.en1867-3880202421Wileyhttps://creativecommons.org/licenses/by-nc-nd/4.0/Enzyme catalysis | Immobilization | Unspecific peroxygenases | Yeast surface displayTechnology::660: Chemistry; Chemical Engineering::660.6: BiotechnologyJournal Articlehttps://doi.org/10.15480/882.1375910.1002/cctc.20240090810.15480/882.13759Journal Article