@article{8091b0943e914c1fa46a2e9775c82e1c,
title = "Spectroelectrochemistry for determination of the redox potential in heme enzymes: Dye-decolorizing peroxidases",
abstract = "Dye-decolorizing peroxidases (DyPs) are heme-containing enzymes that are structurally unrelated to other peroxidases. Some DyPs show high potential for applications in biotechnology, which critically depends on the stability and redox potential (E°') of the enzyme. Here we provide a comparative analysis of UV–Vis- and surface-enhanced resonance Raman-based spectroelectrochemical methods for determination of the E°' of DyPs from two different organisms, and their variants generated targeting E°' upshift. We show that substituting the highly conserved Arginine in the distal side of the heme pocket by hydrophobic amino acid residues impacts the heme architecture and redox potential of DyPs from the two organisms in a very distinct manner. We demonstrate the advantages and drawbacks of the used spectroelectrochemical approaches, which is relevant for other heme proteins that contain multiple heme centers or spin populations.",
keywords = "Dye decolorizing peroxidases, Heme proteins, Redox potential, Spectroelectrochemistry",
author = "Catarina Barbosa and Rodrigues, {Carolina F.} and Nikola Lon{\v c}ar and Martins, {L{\'i}gia O.} and Smilja Todorovic and Silveira, {C{\'e}lia M.}",
note = "Funding Information: The authors thank Dr. Caterina Martin, from GECCO Biotech B.V., for the help in the construction of CboDyP variants and Prof. Marco W. Fraaije for helpful discussions. This work was supported by FCT - Funda{\c c}{\~a}o para a Ci{\^e}ncia e a Tecnologia, I.P., through MOSTMICRO-ITQB R&D Unit ( UIDB/04612/2020 , UIDP/04612/2020 ) and LS4FUTURE Associated Laboratory ( LA/P/0087/2020 ). The authors acknowledge the support from the European Union's Horizon 2020 Research and Innovation Program, through B-LigZymes project (grant agreement number 824017 ) and from COST Action CA21115, supported by COST (European Cooperation in Science and Technology). CB and CRF acknowledge FCT Ph.D. studentships 2020.05017.BD and UI/BD/153388/2022, respectively. Publisher Copyright: {\textcopyright} 2023",
year = "2024",
month = jan,
doi = "10.1016/j.bbadva.2023.100112",
language = "English",
volume = "5",
journal = "BBA Advances",
issn = "2667-1603",
publisher = "Elsevier Science Publisher B.V.",
}