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Contribution of YPRO15C Overexpression to the Resistance of Saccharomyces cerevisiae BY4742 Strain to Furfural Inhibitor

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Polish Journal of Microbiology

Volumen 72 (2023): Edición 2 (June 2023)

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Article Category: ORIGINAL PAPER

Publicado en línea: 14 jun 2023

Páginas: 177 - 186

Recibido: 09 feb 2023

Aceptado: 13 abr 2023

DOI: https://doi.org/10.33073/pjm-2023-019

Palabras clave
furfural, overexpressed, tolerance, transcriptomics

© 2023 Getachew Tafere Abrha et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Construction illustration of the recombinant plasmid. a) pUG6-TEF1p-YPR015C, b) integration diagram of the recombinant plasmid pUG6-TEF1p-YPR into the chromosomal DNA of Saccharomyces cerevisiae.

Cell growth of Saccharomyces cerevisiae strains overexpressing YPR015C and BY4742 as measured at OD600 on a defined synthetic medium containing 35 mM of furfural.
The prolonged lag phase indicates the inhibitory effect of furfural on the parental strain. — Cell growth of Saccharomyces cerevisiae strains overexpressing YPR015C and BY4742 as measured at OD600 on a defined synthetic medium containing 35 mM of furfural. The prolonged lag phase indicates the inhibitory effect of furfural on the parental strain.

Furfural and HMF reduction of activities in the crude cell extracts of the parental (BY4742) and YPR015C overexpressing strains at 35 mM of furfural. The activities were measured using NADH or NADPH as cofactors.
a) NADH and c) NADPH used for furfural reduction, b) and d) for HMF reduction, respectively. The data represent averages of three experiments. *p < 0.05; **p < 0.01 indicates significant differences. — Furfural and HMF reduction of activities in the crude cell extracts of the parental (BY4742) and YPR015C overexpressing strains at 35 mM of furfural. The activities were measured using NADH or NADPH as cofactors. a) NADH and c) NADPH used for furfural reduction, b) and d) for HMF reduction, respectively. The data represent averages of three experiments. *p < 0.05; **p < 0.01 indicates significant differences.

Accumulation of reactive oxygen species (ROS) caused by furfural.
a) Representative images of cells stained with the ROS indicator 2’,7’-dichlorofluorescein diacetate, b) percentage of cells at each concentration of furfural and peroxide that stained positive for ROS by 2’,7’-dichlorofluorescein diacetate after 2, 8, and 16 h. The data represent averages of three experiments with standard error. *p < 0.05; **p < 0.01 indicates significant differences when at least 100 cells were examined on each bright-field image. — Accumulation of reactive oxygen species (ROS) caused by furfural. a) Representative images of cells stained with the ROS indicator 2’,7’-dichlorofluorescein diacetate, b) percentage of cells at each concentration of furfural and peroxide that stained positive for ROS by 2’,7’-dichlorofluorescein diacetate after 2, 8, and 16 h. The data represent averages of three experiments with standard error. *p < 0.05; **p < 0.01 indicates significant differences when at least 100 cells were examined on each bright-field image.

Changes in cell densities after the treatment with lyticase during 4 h of incubation. Saccharomyces cerevisiae strains were treated for 2, 8, and 16 h (from top to bottom) with 35 mM of furfural, respectively. The mean values of relative optical density are presented with vertical error bars, each representing a single standard deviation (n = 3).

eISSN:: 2544-4646
Idioma:: Inglés

Calendario de la edición:: 4 veces al año
Temas de la revista:: Life Sciences, Microbiology and Virology

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