title: Beyond the outer limits of nature by directed evolution.
abstract: For more than thirty years, biotechnology has borne witness to the power of directed evolution in designing molecules of industrial relevance. While scientists all over the world discuss the future of molecular evolution, dozens of laboratory-designed products are being released with improved characteristics in terms of turnover rates, substrate scope, catalytic promiscuity or stability. In this review we aim to present the most recent advances in this fascinating research field that are allowing us to surpass the limits of nature and apply newly gained attributes to a range of applications, from gene therapy to novel green processes. The use of directed evolution in non-natural environments, the generation of catalytic promiscuity for non-natural reactions, the insertion of unnatural amino acids into proteins or the creation of unnatural DNA, is described comprehensively, together with the potential applications in bioremediation, biomedicine and in the generation of new bionanomaterials. These successful case studies show us that the limits of directed evolution will be defined by our own imagination, and in some cases, stretching beyond that.
authors.affiliation: Department of Biocatalysis, Institute of Catalysis, CSIC, Cantoblanco, 28049 Madrid, Spain.
authors.forename: Patricia
authors.initials: P
authors.lastname: Molina-Espeja
authors.affiliation: Department of Biocatalysis, Institute of Catalysis, CSIC, Cantoblanco, 28049 Madrid, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Viña-Gonzalez
authors.affiliation: Department of Biocatalysis, Institute of Catalysis, CSIC, Cantoblanco, 28049 Madrid, Spain.
authors.forename: Bernardo J
authors.initials: BJ
authors.lastname: Gomez-Fernandez
authors.affiliation: Department of Biocatalysis, Institute of Catalysis, CSIC, Cantoblanco, 28049 Madrid, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Martin-Diaz
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, 600 South Mathews Ave, Urbana, IL 61801, USA; Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 600 South Mathews Ave, Urbana, IL 61801, USA.
authors.forename: Eva
authors.initials: E
authors.lastname: Garcia-Ruiz
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: D057005:Bioengineering; D040761:Biomimetic Materials; D001709:Biotechnology; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019020:Directed Molecular Evolution; D049329:Nanostructures; D058615:Synthetic Biology
chemical_list: D004247:DNA
keywords: Biomedicine; Bionanomaterials; Bioremediation; Directed evolution; Gene therapy; Non-natural biocatalysis; Non-natural environments; Unnatural DNA; Unnatural amino acids