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Issue 19, 2019
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Engineering of a fungal laccase to develop a robust, versatile and highly-expressed biocatalyst for sustainable chemistry

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Abstract

Fungal laccases can play an important role as biocatalysts in organic chemistry to replace chemical synthesis. In a previous work we synthesized conductive polyaniline using a high-redox potential laccase from our collection of recombinant fungal variants. Still, the oxidation of aniline is hindered by the reaction conditions (low pH and presence of anionic surfactants). Thus, we tackle here the directed evolution of the enzyme assisted by computational simulation aiming at improving aniline oxidation at the required polymerization conditions while maintaining the enzyme's substrate promiscuity. Simultaneously, its secretion by the host used for the engineering (Saccharomyces cerevisiae) was enhanced. Then, the improved laccase variant was overproduced in the industrial host Aspergillus oryzae and assayed for one-pot synthesis of polyaniline and naphtol-derived dyes whose textile dyeing properties were verified in an industrial environment. Finally, modification of its C-terminal tail further enhanced laccase stability by flexibilization of the region. The resulting biocatalyst displays noticeable stability at high temperature and extreme pH while shows improved kcat values on the different substrates tested. Moreover, it is remarkably produced in S. cerevisiae at rates not formerly reported in the literature. These facts, together with the overexpression in A. oryzae opens new scenarios for its further development and application.

Graphical abstract: Engineering of a fungal laccase to develop a robust, versatile and highly-expressed biocatalyst for sustainable chemistry

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Supplementary files

Article information


Submitted
18 Jul 2019
Accepted
03 Sep 2019
First published
03 Sep 2019

This article is Open Access

Green Chem., 2019,21, 5374-5385
Article type
Paper

Engineering of a fungal laccase to develop a robust, versatile and highly-expressed biocatalyst for sustainable chemistry

F. de Salas, P. Aza, J. F. Gilabert, G. Santiago, S. Kilic, M. E. Sener, J. Vind, V. Guallar, A. T. Martínez and S. Camarero, Green Chem., 2019, 21, 5374
DOI: 10.1039/C9GC02475A

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