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

Getachew Tafere Abrha

Getachew Tafere Abrha

  • Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
  • Department of Biotechnology, College of Dry Land Agriculture and Natural Resources, Mekelle UniversityMekelle, Ethiopia
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Abrha, Getachew Tafere
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Qian Li

Qian Li

Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
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Li, Qian
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Xiaolin Kuang

Xiaolin Kuang

Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
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Kuang, Xiaolin
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Difan Xiao

Difan Xiao

Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
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Xiao, Difan
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Ellen Ayepa

Ellen Ayepa

Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
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Ayepa, Ellen
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Jinjian Wu

Jinjian Wu

Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
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Wu, Jinjian
, 
Huan Chen

Huan Chen

Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
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Chen, Huan
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Zhengyue Zhang

Zhengyue Zhang

Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
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Zhang, Zhengyue
, 
Yina Liu

Yina Liu

Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
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Liu, Yina
, 
Xiumei Yu

Xiumei Yu

Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
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Yu, Xiumei
, 
Quanju Xiang

Quanju Xiang

Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
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Xiang, Quanju
 und 
Menggen Ma

Menggen Ma

  • Department of Applied Microbiology, College of Resources, Sichuan Agricultural UniversitySichuan, China
  • Institute of Natural Resources and Geographic Information Technology, College of Resources, Sichuan Agricultural UniversitySichuan, China
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Ma, Menggen
  
14. Juni 2023
Contribution of YPRO15C Overexpression to the Resistance of Saccharomyces cerevisiae BY4742 Strain to Furfural Inhibitor's Cover Image
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Artikel-Kategorie: ORIGINAL PAPER
Online veröffentlicht: 14. Juni 2023
Seitenbereich: 177 - 186
Eingereicht: 09. Feb. 2023
Akzeptiert: 13. Apr. 2023
DOI: https://doi.org/10.33073/pjm-2023-019
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© 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.
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.

Fig. 1.

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.

Fig. 2.

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.

Fig. 3.

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.

Fig. 4.

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).
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).
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