title: Functional expression, production, and biochemical characterization of a laccase using yeast surface display technology.
abstract: A Trametes versicolor laccase was functionally expressed on the membrane surface of Saccharomyces cerevisiae EBY100. Laccase expression was increased 6.57-fold by medium optimization and surpassed production by the native strain. Maximal laccase and biomass production reached 19 735 ± 1719 Ug-1and 6.22 ± 0.53 gL-1respectively, after 2 d of culture. Optimum oxidization of all substrates by laccase was observed at pH 3. Laccase showed high affinity towards substrates used with Km (mM) and Vmax (μmol min-1) values of 0.57 ± 0.0047 and 24.55 ± 0.64, 1.52 ± 0.52 and 9.25 ± 1.78, and 2.67 ± 0.12 and 11.26 ± 0.75, were reported for ABTS, 2, 6-DMP and GUA, respectively. EDTA and NaN3displayed none competitive inhibition towards laccase activity. The optimum temperature for activity was 50 °C; however, the enzyme was stable over a wide range of temperatures (25-70 °C). The biologically immobilized laccase showed high reusability towards phenolic substrates and low reusability with non-phenolic substrates. High affinity for a diversity phenolic compounds and great ethanol tolerance substantiates this laccase/yeast biocatalyst potential for application in the production of bioethanol.
authors.affiliation: Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, CP 62209, Mexico.
authors.forename: Brandt
authors.initials: B
authors.lastname: Bertrand
authors.affiliation: Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, CP 62209, Mexico.
authors.forename: María R
authors.initials: MR
authors.lastname: Trejo-Hernández
authors.affiliation: Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, CP 62209, Mexico.
authors.forename: Daniel
authors.initials: D
authors.lastname: Morales-Guzmán
authors.affiliation: Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, CP 62209, Mexico.
authors.forename: Luis
authors.initials: L
authors.lastname: Caspeta
authors.affiliation: Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, CP 62209, Mexico.
authors.forename: Ramón
authors.initials: R
authors.lastname: Suárez Rodríguez
authors.affiliation: Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, CP 62209, Mexico. Electronic address: fernandomm@uaem.mx.
authors.forename: Fernando
authors.initials: F
authors.lastname: Martínez-Morales
mesh_terms: D061505:Cell Surface Display Techniques; D003470:Culture Media / Q000737:chemistry; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D042845:Laccase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D055454:Trametes / Q000201:enzymology / Q000235:genetics
chemical_list: D003470:Culture Media; D004800:Enzymes, Immobilized; D042845:Laccase
keywords: Laccase immobilization; Medium optimization; Saccharomyces cerevisiae EBY100; Whole-cell biocatalyst