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

Getachew Tafere Abrha

Abrha, Getachew Tafere

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Zhang, Zhengyue

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14 giu 2023

Contribution of YPRO15C Overexpression to the Resistance of Saccharomyces cerevisiae BY4742 Strain to Furfural Inhibitor's Cover Image

Polish Journal of Microbiology

Volume 72 (2023): Numero 2 (Giugno 2023)

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Categoria dell'articolo: ORIGINAL PAPER

Pubblicato online: 14 giu 2023

Pagine: 177 - 186

Ricevuto: 09 feb 2023

Accettato: 13 apr 2023

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

Parole chiave
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).

Lingua:: Inglese

Frequenza di pubblicazione:: 4 volte all'anno
Argomenti della rivista:: Scienze biologiche, Microbiologia e virologia

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