ArticleScientific reports2017
Evolving stability and pH-dependent activity of the high redox potential Botrytis aclada laccase for enzymatic fuel cells.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 46 citations in OpenAlex.
- A la carte bioprospecting of substrate-selective laccases via high-throughput computational enzyme-substrate interaction profiling.Protein science : a publication of the Protein Society · 2026Article
- Insights into the Role of the D-Cluster in [NiFe]-CODH from Rhodospirillum Rubrum.Chemistry (Weinheim an der Bergstrasse, Germany) · 2025Article
- Computation suggests that the cell adhesion sub-proteome is enriched for sites of pH-dependence and charge burial.PloS one · 2025Article
- A self-regulated network for dynamically balancing multiple precursors in complex biosynthetic pathways.Metabolic engineering · 2024Article
- Fungal Laccases: Fundamentals, Engineering and Classification Update.Biomolecules · 2023Review
- The Physical Basis for pH Sensitivity in Biomolecular Structure and Function, With Application to the Spike Protein of SARS-CoV-2.Frontiers in molecular biosciences · 2022Review
- Heterologous Expression, Engineering and Characterization of a Novel Laccase ofJournal of fungi (Basel, Switzerland) · 2021Article
- Charge Transfer and Biocompatibility Aspects in Conducting Polymer-Based Enzymatic Biosensors and Biofuel Cells.Nanomaterials (Basel, Switzerland) · 2021Review
- Protein Engineering for Improving and Diversifying Natural Product Biosynthesis.Trends in biotechnology · 2020Review
- The first fungal laccase with an alkaline pH optimum obtained by directed evolution and its application in indigo dye decolorization.AMB Express · 2019Article
- Directed Evolution of a Homodimeric Laccase fromInternational journal of molecular sciences · 2018Article
- Improvement of efficiency of brown coal biosolubilization by novel recombinant Fusarium oxysporum laccase.AMB Express · 2018Article
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Authors and funding
13 authors at 3 institutions in 3 countries.
Funding
Abstract
Fungal high redox potential laccases are proposed as cathodic biocatalysts in implantable enzymatic fuel cells to generate high cell voltages. Their application is limited mainly through their acidic pH optimum and chloride inhibition. This work investigates evolutionary and engineering strategies to increase the pH optimum of a chloride-tolerant, high redox potential laccase from the ascomycete Botrytis aclada. The laccase was subjected to two rounds of directed evolution and the clones screened for increased stability and activity at pH 6.5. Beneficial mutation sites were investigated by semi-rational and combinatorial mutagenesis. Fourteen variants were characterised in detail to evaluate changes of the kinetic constants. Mutations increasing thermostability were distributed over the entire structure. Among them, T383I showed a 2.6-fold increased half-life by preventing the loss of the T2 copper through unfolding of a loop. Mutations affecting the pH-dependence cluster around the T1 copper and categorise in three types of altered pH profiles: pH-type I changes the monotonic decreasing pH profile into a bell-shaped profile, pH-type II describes increased specific activity below pH 6.5, and pH-type III increased specific activity above pH 6.5. Specific activities of the best variants were up to 5-fold higher (13 U mg
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