title: Laccase engineering: from rational design to directed evolution.
abstract: Laccases are multicopper oxidoreductases considered by many in the biotechonology field as the ultimate "green catalysts". This is mainly due to their broad substrate specificity and relative autonomy (they use molecular oxygen from air as an electron acceptor and they only produce water as by-product), making them suitable for a wide array of applications: biofuel production, bioremediation, organic synthesis, pulp biobleaching, textiles, the beverage and food industries, biosensor and biofuel cell development. Since the beginning of the 21st century, specific features of bacterial and fungal laccases have been exhaustively adapted in order to reach the industrial demands for high catalytic activity and stability in conjunction with reduced production cost. Among the goals established for laccase engineering, heterologous functional expression, improved activity and thermostability, tolerance to non-natural media (organic solvents, ionic liquids, physiological fluids) and resistance to different types of inhibitors are all challenges that have been met, while obtaining a more comprehensive understanding of laccase structure-function relationships. In this review we examine the most significant advances in this exciting research area in which rational, semi-rational and directed evolution approaches have been employed to ultimately convert laccases into high value-added biocatalysts.
authors.affiliation: DWI-Leibniz-Institute for Interactive Materials, Forckenbeckstraβe 50, 52056, Aachen, Germany.
authors.forename: Diana M
authors.initials: DM
authors.lastname: Mate
authors.affiliation: Department of Biocatalysis, Institute of Catalysis, CSIC, Cantoblanco, 28049 Madrid, Spain. Electronic address: malcalde@icp.csic.es.
authors.forename: Miguel
authors.initials: M
authors.lastname: Alcalde
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D019020:Directed Molecular Evolution; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics; D042845:Laccase / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D015202:Protein Engineering; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D005656:Fungal Proteins; D042845:Laccase
keywords: Bacterial laccase; DNA recombination; Directed evolution; Functional expression; Fungal laccase; Laccase chimeras; Rational design; Redox potential; Saturation mutagenesis