Generation and Characterization of Acid Tolerant Fibrobacter succinogenes S85

Abstract Microorganisms are key components for plant biomass breakdown within rumen environments. Fibrobacter succinogenes have been identified as being active and dominant cellulolytic members of the rumen. In this study, F. succinogenes type strain S85 was adapted for steady state growth in contin...

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Main Authors: Chia-wei Wu, Thomas Spike, Dawn M. Klingeman, Miguel Rodriguez, Virgil R. Bremer, Steven D. Brown
Format: Article
Language:English
Published: Nature Publishing Group 2017-05-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-017-02628-w
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spelling doaj-b6f86f4e710a42288a69e44db6e8bb982020-12-08T02:13:16ZengNature Publishing GroupScientific Reports2045-23222017-05-017111510.1038/s41598-017-02628-wGeneration and Characterization of Acid Tolerant Fibrobacter succinogenes S85Chia-wei Wu0Thomas Spike1Dawn M. Klingeman2Miguel Rodriguez3Virgil R. Bremer4Steven D. Brown5Biosciences Division, Oak Ridge National LaboratoryBiota Biosciences, Inc.Biosciences Division, Oak Ridge National LaboratoryBiosciences Division, Oak Ridge National LaboratoryElanco Animal HealthBiosciences Division, Oak Ridge National LaboratoryAbstract Microorganisms are key components for plant biomass breakdown within rumen environments. Fibrobacter succinogenes have been identified as being active and dominant cellulolytic members of the rumen. In this study, F. succinogenes type strain S85 was adapted for steady state growth in continuous culture at pH 5.75 and confirmed to grow in the range of pH 5.60–5.65, which is lower than has been reported previously. Wild type and acid tolerant strains digested corn stover with equal efficiency in batch culture at low pH. RNA-seq analysis revealed 268 and 829 genes were differentially expressed at pH 6.10 and 5.65 compared to pH 6.70, respectively. Resequencing analysis identified seven single nucleotide polymorphisms (SNPs) in the sufD, yidE, xylE, rlmM, mscL and dosC genes of acid tolerant strains. Due to the absence of a F. succinogenes genetic system, homologues in Escherichia coli were mutated and complemented and the resulting strains were assayed for acid survival. Complementation with wild-type or acid tolerant F. succinogenes sufD restored E. coli wild-type levels of acid tolerance, suggesting a possible role in acid homeostasis. Recent genetic engineering developments need to be adapted and applied in F. succinogenes to further our understanding of this bacterium.https://doi.org/10.1038/s41598-017-02628-w
collection DOAJ
language English
format Article
sources DOAJ
author Chia-wei Wu
Thomas Spike
Dawn M. Klingeman
Miguel Rodriguez
Virgil R. Bremer
Steven D. Brown
spellingShingle Chia-wei Wu
Thomas Spike
Dawn M. Klingeman
Miguel Rodriguez
Virgil R. Bremer
Steven D. Brown
Generation and Characterization of Acid Tolerant Fibrobacter succinogenes S85
Scientific Reports
author_facet Chia-wei Wu
Thomas Spike
Dawn M. Klingeman
Miguel Rodriguez
Virgil R. Bremer
Steven D. Brown
author_sort Chia-wei Wu
title Generation and Characterization of Acid Tolerant Fibrobacter succinogenes S85
title_short Generation and Characterization of Acid Tolerant Fibrobacter succinogenes S85
title_full Generation and Characterization of Acid Tolerant Fibrobacter succinogenes S85
title_fullStr Generation and Characterization of Acid Tolerant Fibrobacter succinogenes S85
title_full_unstemmed Generation and Characterization of Acid Tolerant Fibrobacter succinogenes S85
title_sort generation and characterization of acid tolerant fibrobacter succinogenes s85
publisher Nature Publishing Group
series Scientific Reports
issn 2045-2322
publishDate 2017-05-01
description Abstract Microorganisms are key components for plant biomass breakdown within rumen environments. Fibrobacter succinogenes have been identified as being active and dominant cellulolytic members of the rumen. In this study, F. succinogenes type strain S85 was adapted for steady state growth in continuous culture at pH 5.75 and confirmed to grow in the range of pH 5.60–5.65, which is lower than has been reported previously. Wild type and acid tolerant strains digested corn stover with equal efficiency in batch culture at low pH. RNA-seq analysis revealed 268 and 829 genes were differentially expressed at pH 6.10 and 5.65 compared to pH 6.70, respectively. Resequencing analysis identified seven single nucleotide polymorphisms (SNPs) in the sufD, yidE, xylE, rlmM, mscL and dosC genes of acid tolerant strains. Due to the absence of a F. succinogenes genetic system, homologues in Escherichia coli were mutated and complemented and the resulting strains were assayed for acid survival. Complementation with wild-type or acid tolerant F. succinogenes sufD restored E. coli wild-type levels of acid tolerance, suggesting a possible role in acid homeostasis. Recent genetic engineering developments need to be adapted and applied in F. succinogenes to further our understanding of this bacterium.
url https://doi.org/10.1038/s41598-017-02628-w
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