Vanillyl alcohol oxidase from \(\textit {Diplodia corticola}\)

  • Vanillyl alcohol oxidases (VAOs) belong to the 4-phenol oxidases family and are found predominantly in lignin-degrading ascomycetes. Systematical investigation of the enzyme family at the sequence level resulted in discovery and characterization of the second recombinantly produced VAO member, \(\it Dc\)VAO, from \(\textit {Diplodia corticola}\). Remarkably high activities for 2,6-substituted substrates like 4-allyl-2,6-dimethoxy-phenol (3.5 \(\pm\) 0.02 U \(mg^{−1}\)) or 4-(hydroxymethyl)-2,6-dimethoxyphenol (6.3 \(\pm\) 0.5 U \(mg^{−1}\)) were observed, which could be attributed to a Phe to Ala exchange in the catalytic center. In order to rationalize this rare substrate preference among VAOs, we resurrected and characterized three ancestral enzymes and performed mutagenesis analyses. The results indicate that a Cys/Glu exchange was required to retain activity for \(\gamma\)-hydroxylations and shifted the acceptance towards benzyl ethers (up to 4.0 \(\pm\) 0.1 U \(mg^{−1}\)). Our findings contribute to the understanding of the functionality of VAO enzyme group, and with \(\it Dc\)VAO, we add a new enzyme to the repertoire of ether cleaving biocatalysts.

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Author:Daniel EggerichsGND, Nils WeindorfGND, Maria Laura MascottiORCiDGND, Natalie WelzelGND, Marco Wilhelmus FraaijeGND, Dirk TischlerORCiDGND
Parent Title (English):Journal of biological chemistry
Subtitle (English):residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives
Publisher:Elsevier B.V.
Place of publication:Amsterdam
Document Type:Article
Date of Publication (online):2024/01/30
Date of first Publication:2023/06/08
Publishing Institution:Ruhr-Universität Bochum, Universitätsbibliothek
Tag:Open Access Fonds
Issue:7, Article 104898
First Page:104898-1
Last Page:104898-13
Article Processing Charge funded by the Deutsche Forschungsgemeinschaft (DFG) and the Open Access Publication Fund of Ruhr-Universität Bochum.
Institutes/Facilities:Lehrstuhl für Pflanzenphysiologie, Nachwuchsgruppe Mikrobielle Biotechnologie
Dewey Decimal Classification:Naturwissenschaften und Mathematik / Biowissenschaften, Biologie, Biochemie
open_access (DINI-Set):open_access
faculties:Fakultät für Biologie und Biotechnologie
Licence (English):License LogoCreative Commons - CC BY 4.0 - Attribution 4.0 International