ArticleApplied and environmental microbiology2014
Directed evolution of unspecific peroxygenase from Agrocybe aegerita.
Article in Applied and environmental microbiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 86 papers.
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Who cites it
86 citing papers in PubMed, 246 citations in OpenAlex.
- Promiscuous fungal peroxygenase fromiScience · 2026Article
- Engineering peroxygenases for pharmaceutical applications: recent advances and future perspectives.Chemical science · 2026Review
- Hybrid Chemo-Enzymatic Heterogeneous Catalyst for Green and Enantioselective Hydroxylation.ACS central science · 2026Article
- Biocatalytic Arene Functionalization Using Integrated Unspecific Peroxygenase and Halohydrin Dehalogenase.Chem & bio engineering · 2026Article
- An Aryl Alcohol Oxidase-Peroxygenase Catalyst System Enables Efficient Alcohol to Aldehyde Transformation.Organic process research & development · 2026Article
- Structure, characterisation and application of an unspecific peroxygenase fromRSC chemical biology · 2026Article
- Discovery of Bacterial Unspecific Peroxygenases.Biochemistry · 2026Article
- N-Glycosylation Influences the Heterologous Expression of an Unspecific Peroxygenase From Marasmius rotula in Saccharomyces cerevisiae.Microbial biotechnology · 2026Article
- A green enzymatic route for the biotransformation of naphthalene to phthalic acid.RSC chemical biology · 2026Article
- Unspecific Peroxygenases-Catalyzed Oxidation of Pharmaceutical Compounds Considered Emerging Contaminants.Chembiochem : a European journal of chemical biology · 2026Article
- Enhanced secretion of an Agrocybe aegerita peroxygenase variant in K. phaffii using the native signal peptide.AMB Express · 2026Article
- Computational rational design of unspecific peroxygenase for C-H oxidation.Science advances · 2026Article
- From fungal diversity to antimicrobial innovation: the potential of biotransformation in the era of resistance.Frontiers in microbiology · 2026Review
- Designing an oxidase toolbox for site-directed oxidation of taxanes.Nature communications · 2025Article
- Optimized Machine Learning for Autonomous Enzymatic Reaction Intensification in a Self-Driving Lab.Biotechnology and bioengineering · 2025Article
- Computationally Designed Peroxygenases That Exhibit Diverse and Selective Terpene Oxyfunctionalization.ACS catalysis · 2025Article
- Optimizing Yeast Surface-Displayed Unspecific Peroxygenase Production for Sustainable Biocatalysis.Bioengineering (Basel, Switzerland) · 2025Article
- Synthesis of 19-Hydroxyarachidonic Acid by Fungal Peroxygenases: An Experimental and Computational Study.ChemSusChem · 2025Article
- Identification of improved signal peptides for heterologous expression in Saccharomyces using a screen that exploits Gaussia luciferase.Scientific reports · 2025Article
- Conductive Hairy Particles With Homogeneous and Janus Design as Carrier Materials for the Efficient Immobilization of Unspecific Peroxygenases.Biotechnology journal · 2025Article
26 more citing papers are in PubMed but not listed here.
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Authors and funding
6 authors at 4 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Unspecific peroxygenase (UPO) represents a new type of heme-thiolate enzyme with self-sufficient mono(per)oxygenase activity and many potential applications in organic synthesis. With a view to taking advantage of these properties, we subjected the Agrocybe aegerita UPO1-encoding gene to directed evolution in Saccharomyces cerevisiae. To promote functional expression, several different signal peptides were fused to the mature protein, and the resulting products were tested. Over 9,000 clones were screened using an ad hoc dual-colorimetric assay that assessed both peroxidative and oxygen transfer activities. After 5 generations of directed evolution combined with hybrid approaches, 9 mutations were introduced that resulted in a 3,250-fold total activity improvement with no alteration in protein stability. A breakdown between secretion and catalytic activity was performed by replacing the native signal peptide of the original parental type with that of the evolved mutant; the evolved leader increased functional expression 27-fold, whereas an 18-fold improvement in the kcat/Km value for oxygen transfer activity was obtained. The evolved UPO1 was active and highly stable in the presence of organic cosolvents. Mutations in the hydrophobic core of the signal peptide contributed to enhance functional expression up to 8 mg/liter, while catalytic efficiencies for peroxidative and oxygen transfer reactions were increased by several mutations in the vicinity of the heme access channel. Overall, the directed-evolution platform described is a valuable point of departure for the development of customized UPOs with improved features and for the study of structure-function relationships.
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