Dataset Information

Enhancement of Thermal Resistance by Metal Ions in Thermotolerant Zymomonas mobilis TISTR 548.

ABSTRACT: The thermal resistance of fermenting microbes is a key characteristic of stable fermentation at high temperatures. Therefore, the effects of various metal ions on the growth of Zymomonas mobilis TISTR 548, a thermotolerant ethanologenic bacterium, at a critical high temperature (CHT) were examined. Addition of Mg2+ and K+ increased CHT by 1°C, but the effects of the addition of Mn2+, Ni2+, Co2+, Al3+, Fe3+, and Zn2+ on CHT were negligible. To understand the physiological functions associated with the addition of Mg2+ or K+, cell morphology, intracellular reactive oxygen species (ROS) level, and ethanol productivity were investigated at 39°C (i.e., above CHT). Cell elongation was repressed by Mg2+, but not by K+. Addition of both metals reduced intracellular ROS level, with only K+ showing the highest reduction strength, followed by both metals and only Mg2+. Additionally, ethanol productivity was recovered with the addition of both metals. Moreover, the addition of Mg2+ or K+ at a non-permissive temperature in 26 thermosensitive, single gene-disrupted mutants of Z. mobilis TISTR 548 revealed that several mutants showed metal ion-specific growth improvement. Remarkably, K+ repressed growth of two mutants. These results suggest that K+ and Mg2+ enhance cell growth at CHT via different mechanisms, which involve the maintenance of low intracellular ROS levels.

SUBMITTER: Kosaka T

PROVIDER: S-EPMC7136397 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Enhancement of Thermal Resistance by Metal Ions in Thermotolerant Zymomonas mobilis TISTR 548.

Kosaka Tomoyuki T Nishioka Aya A Sakurada Tomoko T Miura Kento K Anggarini Sakunda S Yamada Mamoru M

Frontiers in microbiology 20200331

The thermal resistance of fermenting microbes is a key characteristic of stable fermentation at high temperatures. Therefore, the effects of various metal ions on the growth of Zymomonas mobilis TISTR 548, a thermotolerant ethanologenic bacterium, at a critical high temperature (CHT) were examined. Addition of Mg2+ and K+ increased CHT by 1°C, but the effects of the addition of Mn2+, Ni2+, Co2+, Al3+, Fe3+, and Z ...[more]

PMID: 32296404

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Project description:UNLABELLED: BACKGROUND:High tolerance to ethanol is a desirable characteristics for ethanologenic strains used in industrial ethanol fermentation. A deeper understanding of the molecular mechanisms underlying ethanologenic strains tolerance of ethanol stress may guide the design of rational strategies to increase process performance in industrial alcoholic production. Many extensive studies have been performed in Saccharomyces cerevisiae and Escherichia coli. However, the physiological basis and genetic mechanisms involved in ethanol tolerance for Zymomonas mobilis are poorly understood on genomic level. To identify the genes required for tolerance to ethanol, microarray technology was used to investigate the transcriptome profiling of the ethanologenic Z. mobilis in response to ethanol stress. RESULTS:We successfully identified 127 genes which were differentially expressed in response to ethanol. Ethanol up- or down-regulated genes related to cell wall/membrane biogenesis, metabolism, and transcription. These genes were classified as being involved in a wide range of cellular processes including carbohydrate metabolism, cell wall/membrane biogenesis, respiratory chain, terpenoid biosynthesis, DNA replication, DNA recombination, DNA repair, transport, transcriptional regulation, some universal stress response, etc. CONCLUSION:In this study, genome-wide transcriptional responses to ethanol were investigated for the first time in Z. mobilis using microarray analysis.Our results revealed that ethanol had effects on multiple aspects of cellular metabolism at the transcriptional level and that membrane might play important roles in response to ethanol. Although the molecular mechanism involved in tolerance and adaptation of ethanologenic strains to ethanol is still unclear, this research has provided insights into molecular response to ethanol in Z. mobilis. These data will also be helpful to construct more ethanol resistant strains for cellulosic ethanol production in the future.

Project description:Zymomonas mobilis ZM4 produces near theoretical yields of ethanol with high specific productivity and recombinant strains are able to ferment both C-5 and C-6 sugars. However, the genetic and physiological basis of the ZM4 response to various industrially-relevant stresses is poorly understood. In this study, the dynamics of ZM4 oxygen stress responses were elucidated by characterizing the transcriptomic and metabolomic profiles of aerobic and anaerobic fermentations using whole-genome microarray analysis and gas chromatography-mass spectrometry. In the absence of oxygen, ZM4 consumed glucose more rapidly, had a higher growth rate, and ethanol was the major end-product. Under aerobic conditions, much less ethanol was observed with other end-products produced, including acetate, lactate, and acetoin. In the early exponential phase, no genes were detected as being significantly differentially expressed between aerobic and anaerobic conditions via microarray analysis. However, microarray analysis of the stationary phase cultures revealed that 166 genes were significantly differentially expressed by more than two-fold. Quantitative-PCR validated the expression values for seventeen genes from different categories of stationary phase microarray data. Transcripts for Entner-Doudoroff pathway genes and gene pdc, encoding a key enzyme leading to the ethanol production, were at least 30-fold more abundant under anaerobic conditions at stationary phase based on quantitative-PCR results. Expression of stress response genes was found to be greater under oxygen stress conditions. GC-MS analysis of stationary phase intracellular metabolites indicated that ZM4 under anaerobic conditions contained lower levels of amino acids, glucose and ED pathway intermediates, whereas metabolites such as ribitol, trehalose, myristic acid, glyceric acid, glucose 6-phosphate, mannose 6-phosphate, and 4-hydroxybutanoic acid were more abundant than under aerobic conditions. Transcriptomic and metabolomic data were consistent with faster glucose consumption and greater ethanol production under anaerobic conditions and suggested gene targets for deletion and improved fermentation. Keywords: Time course/ Stress Response

Dataset Information

Enhancement of Thermal Resistance by Metal Ions in Thermotolerant Zymomonas mobilis TISTR 548.

Publications

Enhancement of Thermal Resistance by Metal Ions in Thermotolerant <i>Zymomonas mobilis</i> TISTR 548.

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