title: Blood tolerant laccase by directed evolution.
abstract: High-redox potential laccases are powerful biocatalysts with a wide range of applications in biotechnology. We have converted a thermostable laccase from a white-rot fungus into a blood tolerant laccase. Adapting the fitness of this laccase to the specific composition of human blood (above neutral pH, high chloride concentration) required several generations of directed evolution in a surrogate complex blood medium. Our evolved laccase was tested in both human plasma and blood, displaying catalytic activity while retaining a high redox potential at the T1 copper site. Mutations introduced in the second coordination sphere of the T1 site shifted the pH activity profile and drastically reduced the inhibitory effect of chloride. This proof of concept that laccases can be adapted to function in extreme conditions opens an array of opportunities for implantable nanobiodevices, chemical syntheses, and detoxification.
authors: Department of Biocatalysis, Institute of Catalysis, CSIC, 28049 Madrid, Spain.
authors: Diana M
authors: DM
authors: Mate
authors: David
authors: D
authors: Gonzalez-Perez
authors: Magnus
authors: M
authors: Falk
authors: Roman
authors: R
authors: Kittl
authors: Marcos
authors: M
authors: Pita
authors: Antonio L
authors: AL
authors: De Lacey
authors: Roland
authors: R
authors: Ludwig
authors: Sergey
authors: S
authors: Shleev
authors: Miguel
authors: M
authors: Alcalde
mesh_terms: D001665:Binding Sites; D002712:Chlorides / Q000737:chemistry; D003300:Copper / Q000737:chemistry; D019020:Directed Molecular Evolution; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D042845:Laccase / Q000097:blood* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D017434:Protein Structure, Tertiary
chemical_list: D002712:Chlorides; D004791:Enzyme Inhibitors; D003300:Copper; D042845:Laccase