Evolved Fusarium oxysporum laccase expressed in Saccharomyces cerevisiae

• Published in: Scientific reports Vol. 10; no. 1; p. 3244
• Main Authors: Kwiatos, Natalia, Jędrzejczak-Krzepkowska, Marzena, Krzemińska, Agnieszka, Delavari, Azar, Paneth, Piotr, Bielecki, Stanisław
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 24-02-2020; Nature Publishing Group; Nature Portfolio
• Subjects: 631/114; 631/114/469; 631/114/663; 631/45; 631/45/535; 631/45/535/1267; 631/45/603; 631/45/607; 631/45/607/1168; 631/45/612; 631/45/612/1141; 631/535; 631/535/1267; 631/61; 631/61/168; 631/61/185; 631/61/338; 631/61/338/469; Coal; Enzymes; Evolution, Molecular; Fulvic acids; Fusarium - enzymology; Fusarium oxysporum; Humanities and Social Sciences; Humic Substances - analysis; Kinetics; Laccase; Laccase - genetics; Laccase - isolation & purification; Laccase - metabolism; Lignin; Molecular Docking Simulation; Molecular Dynamics Simulation; multidisciplinary; Mutation - genetics; Protein Engineering; Saccharomyces cerevisiae; Saccharomyces cerevisiae - metabolism; Science; Science (multidisciplinary); Thermodynamics; Yeast
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-60204-1

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.