Production and manipulation of blue copper oxidases for technological applications.

Autor: Zhou S; Aix Marseille Université, CNRS, Centrale Marseille, iSm2, Marseille, France., Rousselot-Pailley P; Aix Marseille Université, CNRS, Centrale Marseille, iSm2, Marseille, France., Ren L; Aix Marseille Université, CNRS, Centrale Marseille, iSm2, Marseille, France., Charmasson Y; Aix Marseille Université, CNRS, Centrale Marseille, iSm2, Marseille, France., Dezord EC; Aix Marseille Université, CNRS, Centrale Marseille, iSm2, Marseille, France., Robert V; Aix Marseille Université, CNRS, Centrale Marseille, iSm2, Marseille, France., Tron T; Aix Marseille Université, CNRS, Centrale Marseille, iSm2, Marseille, France., Mekmouche Y; Aix Marseille Université, CNRS, Centrale Marseille, iSm2, Marseille, France. Electronic address: y.mekmouche@univ-amu.fr.
Jazyk: angličtina
Zdroj: Methods in enzymology [Methods Enzymol] 2018; Vol. 613, pp. 17-61. Date of Electronic Publication: 2018 Nov 24.
DOI: 10.1016/bs.mie.2018.10.015
Abstrakt: Fungal laccases are robust multicopper oxidoreductases. Perfectly amenable to synthetic evolution, the fungal laccase scaffold is a potential generic for the production of tailored biocatalysts, which, in principle, can be secreted at substantial levels in industrially relevant organisms. In this chapter, the strategy we have developed for the rapid production of hundreds of milligram of laccase variants is detailed. It is based on the use of two heterologous expression hosts: the yeast Saccharomyces cerevisiae for a rapid upstream screening and the fungus Aspergillus niger for downstream production. Methods for screening active and nonactive laccase variants, convenient setups for enzyme production in both organisms as well as a methodology for efficient purification of large amounts of recombinant enzymes are given. The general procedure for developing new materials for artificial catalysis is also described.
(© 2018 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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