Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae

Abstract Fusarium oxysporum laccase was functionally expressed in Saccharomyces cerevisiae and engineered towards higher expression levels and higher reactivity towards 2,6-dimethoxyphenol, that could be used as a mediator for lignin modification. A combination of classical culture optimization and...

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Autores principales:	Natalia Kwiatos, Marzena Jędrzejczak-Krzepkowska, Agnieszka Krzemińska, Azar Delavari, Piotr Paneth, Stanisław Bielecki
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3c2feebd20c049e298f63f8604c1a474
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spelling	oai:doaj.org-article:3c2feebd20c049e298f63f8604c1a4742021-12-02T14:28:22ZEvolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae10.1038/s41598-020-60204-12045-2322https://doaj.org/article/3c2feebd20c049e298f63f8604c1a4742020-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-60204-1https://doaj.org/toc/2045-2322Abstract Fusarium oxysporum laccase was functionally expressed in Saccharomyces cerevisiae and engineered towards higher expression levels and higher reactivity towards 2,6-dimethoxyphenol, that could be used as a mediator for lignin modification. A combination of classical culture optimization and protein engineering led to around 30 times higher activity in the culture supernatant. The winner mutant exhibited three times lower Km, four times higher kcat and ten times higher catalytic efficiency than the parental enzyme. The strategy for laccase engineering was composed of a combination of random methods with a rational approach based on QM/MM MD studies of the enzyme complex with 2,6-dimethoxyphenol. Laccase mediator system with 2,6-dimethoxyphenol caused fulvic acids release from biosolubilized coal.Natalia KwiatosMarzena Jędrzejczak-KrzepkowskaAgnieszka KrzemińskaAzar DelavariPiotr PanethStanisław BieleckiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-11 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Natalia Kwiatos Marzena Jędrzejczak-Krzepkowska Agnieszka Krzemińska Azar Delavari Piotr Paneth Stanisław Bielecki Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae
description	Abstract Fusarium oxysporum laccase was functionally expressed in Saccharomyces cerevisiae and engineered towards higher expression levels and higher reactivity towards 2,6-dimethoxyphenol, that could be used as a mediator for lignin modification. A combination of classical culture optimization and protein engineering led to around 30 times higher activity in the culture supernatant. The winner mutant exhibited three times lower Km, four times higher kcat and ten times higher catalytic efficiency than the parental enzyme. The strategy for laccase engineering was composed of a combination of random methods with a rational approach based on QM/MM MD studies of the enzyme complex with 2,6-dimethoxyphenol. Laccase mediator system with 2,6-dimethoxyphenol caused fulvic acids release from biosolubilized coal.
format	article
author	Natalia Kwiatos Marzena Jędrzejczak-Krzepkowska Agnieszka Krzemińska Azar Delavari Piotr Paneth Stanisław Bielecki
author_facet	Natalia Kwiatos Marzena Jędrzejczak-Krzepkowska Agnieszka Krzemińska Azar Delavari Piotr Paneth Stanisław Bielecki
author_sort	Natalia Kwiatos
title	Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae
title_short	Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae
title_full	Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae
title_fullStr	Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae
title_full_unstemmed	Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae
title_sort	evolved fusarium oxysporum laccase expressed in saccharomyces cerevisiae
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/3c2feebd20c049e298f63f8604c1a474
work_keys_str_mv	AT nataliakwiatos evolvedfusariumoxysporumlaccaseexpressedinsaccharomycescerevisiae AT marzenajedrzejczakkrzepkowska evolvedfusariumoxysporumlaccaseexpressedinsaccharomycescerevisiae AT agnieszkakrzeminska evolvedfusariumoxysporumlaccaseexpressedinsaccharomycescerevisiae AT azardelavari evolvedfusariumoxysporumlaccaseexpressedinsaccharomycescerevisiae AT piotrpaneth evolvedfusariumoxysporumlaccaseexpressedinsaccharomycescerevisiae AT stanisławbielecki evolvedfusariumoxysporumlaccaseexpressedinsaccharomycescerevisiae
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Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae

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