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Cellular Analysis and Comparative Transcriptomics Reveal the Tolerance Mechanisms of Candida tropicalis Toward Phenol.

ABSTRACT: Phenol is a ubiquitous pollutant and can contaminate natural water resources. Hence, the removal of phenol from wastewater is of significant importance. A series of biological methods were used to remove phenol based on the natural ability of microorganisms to degrade phenol, but the tolerance mechanism of phenol-degraded strains to phenol are not very clear. Morphological observation on Candida tropicalis showed that phenol caused the reactive oxygen species (ROS) accumulation, damaging the mitochondrial and the endoplasmic reticulum. On the basis of transcriptome data and cell wall susceptibility analysis, it was found that C. tropicalis prevented phenol-caused cell damage through improvement of cell wall resistance, maintenance of high-fidelity DNA replication, intracellular protein homeostasis, organelle integrity, and kept the intracellular phenol concentration at a low level through cell-wall remodeling and removal of excess phenol via MDR/MXR transporters. The knowledge obtained will promote the genetic modification of yeast strains in general to tolerate the high concentrations of phenol and improve their efficiency of phenol degradation.

SUBMITTER: Wang H

PROVIDER: S-EPMC7179700 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Cellular Analysis and Comparative Transcriptomics Reveal the Tolerance Mechanisms of <i>Candida tropicalis</i> Toward Phenol.

Wang Hanyu H Li Qian Q Peng Yuanyuan Y Zhang Zhengyue Z Kuang Xiaolin X Hu Xiangdong X Ayepa Ellen E Han Xuebing X Abrha Getachew Tafere GT Xiang Quanju Q Yu Xiumei X Zhao Ke K Zou Likou L Gu Yunfu Y Li Xi X Li Xiaoying X Chen Qiang Q Zhang Xiaoping X Liu Beidong B Ma Menggen M

Frontiers in microbiology 20200415

Phenol is a ubiquitous pollutant and can contaminate natural water resources. Hence, the removal of phenol from wastewater is of significant importance. A series of biological methods were used to remove phenol based on the natural ability of microorganisms to degrade phenol, but the tolerance mechanism of phenol-degraded strains to phenol are not very clear. Morphological observation on <i>Candida tropicalis</i> showed that phenol caused the reactive oxygen species (ROS) accumulation, damaging ...[more]

PMID: 32373081

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Project description:BackgroundCurrent biomass pretreatment by hydrothermal treatment (including acid hydrolysis, steam explosion, and high-temperature steaming) and ionic liquids generally generate inhibitors to the following fermentation process. Furfural is one of the typical inhibitors generated in hydrothermal treatment of biomass. Furfural could inhibit cell growth rate and decrease biofuel productivity of microbes. Candida tropicalis is a promising microbe for the production of biofuels and value-added chemicals using hemicellulose hydrolysate as carbon source. In this study, C. tropicalis showed a comparable ability of furfural tolerance during fermentation. We investigated the mechanism of C. tropicalis's robust tolerance to furfural and relevant metabolic responses to obtain more information for metabolic engineering of microbes for efficient lignocellulose fermentation.ResultsCandida tropicalis showed comparable intrinsic tolerance to furfural and a fast rate of furfural detoxification. C. tropicalis's half maximal inhibitory concentration for furfural with xylose as the sole carbon source was 3.69 g/L, which was higher than that of most wild-type microbes reported in the literature to our knowledge. Even though furfural prolonged the lag phase of C. tropicalis, the final biomass in the groups treated with 1 g/L furfural was slightly greater than that in the control groups. By real-time PCR analysis, we found that the expression of ADH1 in C. tropicalis (ctADH1) was induced by furfural and repressed by ethanol after furfural depletion. The expression of ctADH1 could be regulated by both furfural and ethanol. After the disruption of gene ctADH1, we found that C. tropicalis's furfural tolerance was weakened. To further confirm the function of ctADH1 and enhance Escherichia coli's furfural tolerance, ctADH1 was overexpressed in E. coli BL21 (DE3). The rate of furfural degradation in E. coli BL21 (DE3) with pET-ADH1 (high-copy plasmid) and pCS-ADH1 (medium-copy plasmid) was increased by 1.59-fold and 1.28-fold, respectively.ConclusionsCandida tropicalis was a robust strain with intrinsic tolerance to inhibitor furfural. The mechanism of furfural detoxification and metabolic responses were identified by multiple analyses. Alcohol dehydrogenase 1 was confirmed to be responsible for furfural detoxification. C. tropicalis showed a complex regulation system during furfural detoxification to minimize adverse effects caused by furfural. Furthermore, the mechanism we uncovered in this work was successfully applied to enhance E. coli's furfural tolerance by heterologous expression of ctADH1. The study provides deeper insights into strain modification for biofuel production by efficient lignocellulose fermentation.

Dataset Information

Cellular Analysis and Comparative Transcriptomics Reveal the Tolerance Mechanisms of Candida tropicalis Toward Phenol.

Publications

Cellular Analysis and Comparative Transcriptomics Reveal the Tolerance Mechanisms of <i>Candida tropicalis</i> Toward Phenol.

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