Next Article in Journal
Hydroxytyrosol Promotes the Mitochondrial Function through Activating Mitophagy
Next Article in Special Issue
Engineering Collariella virescens Peroxygenase for Epoxides Production from Vegetable Oil
Previous Article in Journal
Halogenation Activity of Mammalian Heme Peroxidases
Previous Article in Special Issue
Novel Fatty Acid Chain-Shortening by Fungal Peroxygenases Yielding 2C-Shorter Dicarboxylic Acids
Article

Structural Characterization of Two Short Unspecific Peroxygenases: Two Different Dimeric Arrangements

1
Centro de Investigaciones Biológicas “Margarita Salas” (CIB), CSIC, E-28040 Madrid, Spain
2
Intravacc, 3721 MA De Bilt, The Netherlands
3
Instituto de Recursos Naturales y Agrobiología de Sevilla (IRNAS), CSIC, E-41012 Seville, Spain
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Alessandra Napolitano
Antioxidants 2022, 11(5), 891; https://doi.org/10.3390/antiox11050891
Received: 25 March 2022 / Revised: 26 April 2022 / Accepted: 28 April 2022 / Published: 30 April 2022
(This article belongs to the Special Issue Dream Peroxygenases)
Unspecific peroxygenases (UPOs) are extracellular fungal enzymes of biotechnological interest as self-sufficient (and more stable) counterparts of cytochrome P450 monooxygenases, the latter being present in most living cells. Expression hosts and structural information are crucial for exploiting UPO diversity (over eight thousand UPO-type genes were identified in sequenced genomes) in target reactions of industrial interest. However, while many thousands of entries in the Protein Data Bank include molecular coordinates of P450 enzymes, only 19 entries correspond to UPO enzymes, and UPO structures from only two species (Agrocybe aegerita and Hypoxylon sp.) have been published to date. In the present study, two UPOs from the basidiomycete Marasmius rotula (rMroUPO) and the ascomycete Collariella virescens (rCviUPO) were crystallized after sequence optimization and Escherichia coli expression as active soluble enzymes. Crystals of rMroUPO and rCviUPO were obtained at sufficiently high resolution (1.45 and 1.95 Å, respectively) and the corresponding structures were solved by molecular replacement. The crystal structures of the two enzymes (and two mutated variants) showed dimeric proteins. Complementary biophysical and molecular biology studies unveiled the diverse structural bases of the dimeric nature of the two enzymes. Intermolecular disulfide bridge and parallel association between two α-helices, among other interactions, were identified at the dimer interfaces. Interestingly, one of the rCviUPO variants incorporated the ability to produce fatty acid diepoxides—reactive compounds with valuable cross-linking capabilities—due to removal of the enzyme C-terminal tail located near the entrance of the heme access channel. In conclusion, different dimeric arrangements could be described in (short) UPO crystal structures. View Full-Text
Keywords: unspecific peroxygenases (UPO); Collariella virescens; Marasmius rotula; crystal structures; analytical ultracentrifugation; dimeric arrangements; fatty acid epoxidation unspecific peroxygenases (UPO); Collariella virescens; Marasmius rotula; crystal structures; analytical ultracentrifugation; dimeric arrangements; fatty acid epoxidation
Show Figures

Figure 1

MDPI and ACS Style

Linde, D.; Santillana, E.; Fernández-Fueyo, E.; González-Benjumea, A.; Carro, J.; Gutiérrez, A.; Martínez, A.T.; Romero, A. Structural Characterization of Two Short Unspecific Peroxygenases: Two Different Dimeric Arrangements. Antioxidants 2022, 11, 891. https://doi.org/10.3390/antiox11050891

AMA Style

Linde D, Santillana E, Fernández-Fueyo E, González-Benjumea A, Carro J, Gutiérrez A, Martínez AT, Romero A. Structural Characterization of Two Short Unspecific Peroxygenases: Two Different Dimeric Arrangements. Antioxidants. 2022; 11(5):891. https://doi.org/10.3390/antiox11050891

Chicago/Turabian Style

Linde, Dolores, Elena Santillana, Elena Fernández-Fueyo, Alejandro González-Benjumea, Juan Carro, Ana Gutiérrez, Angel T. Martínez, and Antonio Romero. 2022. "Structural Characterization of Two Short Unspecific Peroxygenases: Two Different Dimeric Arrangements" Antioxidants 11, no. 5: 891. https://doi.org/10.3390/antiox11050891

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop