Functional expression, production, and biochemical characterization of a laccase using yeast surface display technology
2016; Elsevier BV; Volume: 120; Issue: 12 Linguagem: Inglês
10.1016/j.funbio.2016.08.009
ISSN1878-6162
AutoresBrandt Bertrand, María R. Trejo‐Hernández, Daniel Morales‐Guzmán, Luis Caspeta, Ramón Suárez‐Rodríguez, Fernando Martínez‐Morales,
Tópico(s)Electrochemical sensors and biosensors
ResumoA 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−1 and 6.22 ± 0.53 gL−1 respectively, 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 NaN3 displayed 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.
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