Artigo Revisado por pares

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

ISSN

1878-6162

Autores

Brandt Bertrand, María R. Trejo‐Hernández, Daniel Morales‐Guzmán, Luis Caspeta, Ramón Suárez‐Rodríguez, Fernando Martínez‐Morales,

Tópico(s)

Electrochemical sensors and biosensors

Resumo

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−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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