ArticleAMB Express2019
The first fungal laccase with an alkaline pH optimum obtained by directed evolution and its application in indigo dye decolorization.
Article in AMB Express, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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39 citing papers in PubMed, 99 citations in OpenAlex.
- A laccase-mediator system based on caffeic acid for the synthesis of benzimidazoles.Scientific reports · 2026Article
- Bio decomposition of paddy straw waste using laccase producing novel soil bacterial isolates: An eco-friendly and sustainable approach.Scientific reports · 2026Article
- Ligninolytic Enzyme Content of MushroomFood science & nutrition · 2026Article
- Adaptive evolution of Agaricomycetes laccases follows wood lignin diversification in plants.Applied and environmental microbiology · 2026Article
- Harnessing the potential role of lignocellulolytic microbial consortia for rapid decomposition of rice straw.Frontiers in microbiology · 2026Article
- Discovery and engineering of bifunctional enzymes for lignocellulose degradation: Metagenomic and computational approaches.Biotechnology reports (Amsterdam, Netherlands) · 2025Review
- Enhancing the Substrate Adaptability of Laccase through Ancestral Sequence Reconstruction for Applications in Mycotoxin Detoxification.Journal of agricultural and food chemistry · 2025Article
- Recent Advances in Laccase Production: Challenges and Future Perspectives.Journal of microbiology and biotechnology · 2025Review
- Specific Phenylpropanoid Oligomerization in a Neutral Environment by the Recombinant Alkaline Laccase fromBiomolecules · 2025Article
- Catalytic Residue Reprogramming Enhances Enzyme Activity at Alkaline pH via Phenolate-Mediated Proton Transfer.ACS synthetic biology · 2025Article
- An Economical Industrially Feasible Approach for Denim Biobleaching: Reusing of Laccase and Mediator for Multiple Cycles.Indian journal of microbiology · 2025Article
- Efficient Decolorization of the Poly-Azo Dye Sirius Grey byMolecules (Basel, Switzerland) · 2024Article
- Role of microbial laccases in valorization of lignocellulosic biomass to bioethanol.Frontiers in bioengineering and biotechnology · 2024Review
- Electricity generation and oxidoreductase potential during dye discoloration by laccase-producing Ganoderma gibbosum in fungal fuel cell.Microbial cell factories · 2023Article
- Fungal Laccases: Fundamentals, Engineering and Classification Update.Biomolecules · 2023Review
- Article
- Effective biosynthesis of 2,5-furandicarboxylic acid from 5-hydroxymethylfurfural via a bi-enzymatic cascade system using bacterial laccase and fungal alcohol oxidase.Biotechnology for biofuels and bioproducts · 2023Article
- Structural Insight into the Amino Acid Environment of the Two-Domain Laccase's Trinuclear Copper Cluster.International journal of molecular sciences · 2023Article
- Characterization of a New Laccase fromMolecules (Basel, Switzerland) · 2023Article
- Biosensors Based on Phenol Oxidases (Laccase, Tyrosinase, and Their Mixture) for Estimating the Total Phenolic Index in Food-Related Samples.Life (Basel, Switzerland) · 2023Review
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8 authors at 2 institutions in 2 countries.
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Abstract
Engineering of fungal laccases with optimum catalytic activity at alkaline pH has been a long-lasting challenge. In this study, a mutant library containing 3000 clones was obtained by error-prone PCR to adapt the optimum pH of a fungal laccase Lcc9 from the basidiomycete Coprinopsis cinerea. After three rounds of functional screening, a mutant with three amino acid changes (E116K, N229D, I393T) named PIE5 was selected. PIE5 showed an optimum pH of 8.5 and 8.0 against guaiacol and 2,6-DMP when expressed in Pichia pastoris, representing the first fungal laccase that possesses an optimum pH at an alkaline condition. Site directed mutagenesis disclosed that N229D contributed the most to the optimum pH increment. A single N229D mutation caused an increase in optimum pH by 1.5 units. When used in indigo dye decolorization, PIE5 efficiently decolorized 87.1 ± 1.1% and 90.9 ± 0.3% indigo dye at the optimum conditions of pH 7.0-7.5 and 60 °C, and with either methyl 3,5-dimethoxy-4-hydroxybenzoate or 2,2'-azino-bis(3-ethylbenzothazoline-6-sulfonate) as the mediator. In comparison, the commercially available fungal laccase TvLac from Trametes villosa decolorized 84.3 ± 1.8% of indigo dye under its optimum conditions (opt. pH 5.0 and 60 °C). The properties of an alkaline-dependent activity and the high indigo dye decolorization ability (1.3-fold better than the parental Lcc9) make the new fungal laccase PIE5 an alternative for specific industrial applications.
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