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Grant support

This work was supported by the European Union [FP7-KBBE-2013-7-613549-INDOX, H2020-BBI-PPP-2015-2-720297-ENZOX2], the COST Action [CM1303 Systems Biocatalysis] and the Spanish Government [BIO2016-79106-R-Lignolution].

Analysis of institutional authors

Mate, Diana M.Author

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November 25, 2019
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Modification of the peroxygenative: peroxidative activity ratio in the unspecific peroxygenase from Agrocybe aegerita by structure-guided evolution

Publicated to:PROTEIN ENGINEERING DESIGN & SELECTION. 30 (3): 191-198 - 2017-01-01 30(3), DOI: 10.1093/protein/gzw073

Authors: Mate, Diana M.; Palomino, Miguel A.; Molina-Espeja, Patricia; Martin-Diaz, Javier; Alcalde, Miguel;

Affiliations

CSIC, Inst Catalysis, Dept Biocatalysis, Marie Curie 2, Madrid 28049, Spain - Author

Abstract

Unspecific peroxygenase (UPO) is a heme-thiolate peroxidase capable of performing with high-selectivity C-H oxyfunctionalizations of great interest in organic synthesis through its peroxygenative activity. However, the convergence of such activity with an unwanted peroxidative activity encumbers practical applications. In this study, we have modified the peroxygenative: peroxidative activity ratio (P:p ratio) of UPO from Agrocybe aegerita by structure-guided evolution. Several flexible loops (Glu1-Pro35, Gly103-Asp131, Ser226-Gly243, Gln254-Thr276 and Ty293-Arg327) were selected on the basis on their B-factors and Delta Delta G values. The full ensemble of segments (43% of UPO sequence) was subjected to focused evolution by the Mutagenic Organized Recombination Process by Homologous IN vivo Grouping (MORPHING) method in Saccharomyces cerevisiae. Five independent mutant libraries were screened in terms of P: p ratio and thermostability. We identified several variants that harbored substitutions at positions 120 and 320 with a strong enhancement in the P: p ratio albeit at the cost of stability. The most thermostable mutant of this process (S226G with an increased T-50 of 2 degrees C) was subjected to further combinatorial saturation mutagenesis on Thr120 and Thr320 yielding a collection of variants with modified P: p ratio and recovered stability. Our results seem to indicate the coexistence of several oxidation sites for peroxidative and peroxygenative activities in UPO.

Keywords

peroxidative activityperoxygenative activitystructure-guided evolutionthermostabilityOxidationPeroxidative activityPeroxygenative activityPopmusicSitesStability changesStructure-guided evolutionThermostabilityUnspecific peroxygenaseVersatile peroxidase

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal PROTEIN ENGINEERING DESIGN & SELECTION due to its progression and the good impact it has achieved in recent years, according to the agency Scopus (SJR), it has become a reference in its field. In the year of publication of the work, 2017, it was in position , thus managing to position itself as a Q1 (Primer Cuartil), in the category Biotechnology.

From a relative perspective, and based on the normalized impact indicator calculated from the Field Citation Ratio (FCR) of the Dimensions source, it yields a value of: 2.96, which indicates that, compared to works in the same discipline and in the same year of publication, it ranks as a work cited above average. (source consulted: Dimensions Jul 2025)

Specifically, and according to different indexing agencies, this work has accumulated citations as of 2025-07-20, the following number of citations:

  • WoS: 25
  • Scopus: 29
  • Europe PMC: 1

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-07-20:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 75.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 76 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 3.

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.

Leadership analysis of institutional authors

There is a significant leadership presence as some of the institution’s authors appear as the first or last signer, detailed as follows: First Author (MATE MATE, DIANA) .