Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae

Author:

Kwiatos Natalia,Jędrzejczak-Krzepkowska Marzena,Krzemińska Agnieszka,Delavari Azar,Paneth Piotr,Bielecki Stanisław

Abstract

AbstractFusarium 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.

Funder

Narodowym Centrum Nauki

Publisher

Springer Science and Business Media LLC

Subject

Multidisciplinary

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