Influence of Osmotic Stress on Fermentative Production of Succinic Acid by Actinobacillus succinogenes

Xiaojiang, Fang; Li, Jian; Zheng, Xiaoyu; Yao, Xi; Chen, Kequan; Wei, Ping; Ouyang, Pingkai; Jiang, Min

doi:10.1007/s12010-011-9239-6

Cited by 15 publications

(3 citation statements)

References 25 publications

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“…Supplementation with betaine increased the cell growth rate, glucose consumption rate, and succinate productivity of engineered E. coli strains in a high initial glucose concentration (150 g/liter) (19). Supplementation with proline increased the succinate titer by 22% from engineered Actinobacillus succinogenes under high osmotic pressure (20). The expression of exogenous betaine biosynthesis genes from the extreme halophile Ectothiorhodospira halochloris resulted in a recombinant E. coli with double the cellular yield compared to that of its control strain in 0.5 M NaCl (21).…”

Section: Resultsmentioning

confidence: 99%

Osmotolerance in Escherichia coli Is Improved by Activation of Copper Efflux Genes or Supplementation with Sulfur-Containing Amino Acids

Xiao

Zhu

Feng

et al. 2017

Appl Environ Microbiol

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Improvement in the osmotolerance of Escherichia coli is essential for the production of high titers of various bioproducts. In this work, a cusS mutation that was identified in the previously constructed high-succinate-producing E. coli strain HX024 was investigated for its effect on osmotolerance. CusS is part of the twocomponent system CusSR that protects cells from Ag(I) and Cu(I) toxicity. Changing cusS from strain HX024 back to its original sequence led to a 24% decrease in cell mass and succinate titer under osmotic stress (12% glucose). When cultivated with a high initial glucose concentration (12%), introduction of the cusS mutation into parental strain Suc-T110 led to a 21% increase in cell mass and a 40% increase in succinate titer. When the medium was supplemented with 30 g/liter disodium succinate, the cusS mutation led to a 120% increase in cell mass and a 492% increase in succinate titer. Introducing the cusS mutation into the wild-type strain ATCC 8739 led to increases in cell mass of 87% with 20% glucose and 36% using 30 g/liter disodium succinate. The cusS mutation increased the expression of cusCFBA, and gene expression levels were found to be positively related to osmotolerance abilities. Because high osmotic stress has been associated with deleterious accumulation of Cu(I) in the periplasm, activation of CusCFBA may alleviate this effect by transporting Cu(I) out of the cells. This hypothesis was confirmed by supplementing sulfurcontaining amino acids that can chelate Cu(I). Adding methionine or cysteine to the medium increased the osmotolerance of E. coli under anaerobic conditions. IMPORTANCE In this work, an activating Cus copper efflux system was found to increase the osmotolerance of E. coli. In addition, new osmoprotectants were identified. Supplementation with methionine or cysteine led to an increase in osmotolerance of E. coli under anaerobic conditions. These new strategies for improving osmotolerance will be useful for improving the production of chemicals in industrial bioprocesses.

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Section: Resultsmentioning

confidence: 99%

Osmotolerance in Escherichia coli Is Improved by Activation of Copper Efflux Genes or Supplementation with Sulfur-Containing Amino Acids

Xiao

Zhu

Feng

et al. 2017

Appl Environ Microbiol

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“…After challenge at pH 4.0 for 5 H, the survival rate of L. lactis NZ9000 (Leu+) increased 28.5‐fold compared to L. lactis NZ9000 (Leu − ). In addition, the succinic acid production of Actinobacillus succinogenes NJ113 was increased by 22.2% with the addition of 25 mM proline . Furthermore, the addition of 1 mM glycine betaine to the growth medium partially relieved osmotic pressure stress in Desulfovibrio halophilus .…”

Section: Process Engineering To Improve Microbial Stress Resistancementioning

confidence: 97%

Techniques for enhancing the tolerance of industrial microbes to abiotic stresses: A review

Zhou

Zhang

et al. 2019

Biotech and App Biochem

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The diversity of stress responses and survival strategies evolved by microorganism enables them to survive and reproduce in a multitude of harsh environments, whereas the discovery of the underlying resistance genes or mechanisms laid the foundation for the directional enhancement of microbial tolerance to abiotic stresses encountered in industrial applications. Many biological techniques have been developed for improving the stress resistance of industrial microorganisms, which greatly benefited the bacteria on which industrial production is based. This review introduces the main techniques for enhancing the resistance of microorganisms to abiotic stresses, including evolutionary engineering, metabolic engineering, and process engineering, developed in recent years. In addition, we also discuss problems that are still present in this area and offer directions for future research.

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“…Compatible solutes are usually impermeable to the cell membrane, less toxic at high internal concentrations, and not easily catabolized [ 6 , 8 ], which greatly facilitates water remaining within the cytoplasm. In terms of SA production, it was reported that medium supplemented with glycine-betaine or proline improved cell osmotolerance and succinate production in E. coli [ 12 ] and Actinobacillus succinogenes [ 13 ]. However, it is worth noting that the osmoprotective effects of these compatible solutes are conditional.…”

Section: Introductionmentioning

confidence: 99%

A novel point mutation in RpoB improves osmotolerance and succinic acid production in Escherichia coli

Xiao

Zhu

et al. 2017

BMC Biotechnol

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Background Escherichia coli suffer from osmotic stress during succinic acid (SA) production, which reduces the performance of this microbial factory.ResultsHere, we report that a point mutation leading to a single amino acid change (D654Y) within the β-subunit of DNA-dependent RNA polymerase (RpoB) significantly improved the osmotolerance of E. coli. Importation of the D654Y mutation of RpoB into the parental strain, Suc-T110, increased cell growth and SA production by more than 40% compared to that of the control under high glucose osmolality. The transcriptome profile, determined by RNA-sequencing, showed two distinct stress responses elicited by the mutated RpoB that counterbalanced the osmotic stress. Under non-stressed conditions, genes involved in the synthesis and transport of compatible solutes such as glycine-betaine, glutamate or proline were upregulated even without osmotic stimulation, suggesting a “pre-defense” mechanism maybe formed in the rpoB mutant. Under osmotic stressed conditions, genes encoding diverse sugar transporters, which should be down-regulated in the presence of high osmotic pressure, were derepressed in the rpoB mutant. Additional genetic experiments showed that enhancing the expression of the mal regulon, especially for genes that encode the glycoporin LamB and maltose transporter, contributed to the osmotolerance phenotype.ConclusionsThe D654Y single amino acid substitution in RpoB rendered E. coli cells resistant to osmotic stress, probably due to improved cell growth and viability via enhanced sugar uptake under stressed conditions, and activated a potential “pre-defense” mechanism under non-stressed conditions. The findings of this work will be useful for bacterial host improvement to enhance its resistance to osmotic stress and facilitate bio-based organic acids production.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-017-0337-6) contains supplementary material, which is available to authorized users.

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Influence of Osmotic Stress on Fermentative Production of Succinic Acid by Actinobacillus succinogenes

Cited by 15 publications

References 25 publications

Osmotolerance in Escherichia coli Is Improved by Activation of Copper Efflux Genes or Supplementation with Sulfur-Containing Amino Acids

Osmotolerance in Escherichia coli Is Improved by Activation of Copper Efflux Genes or Supplementation with Sulfur-Containing Amino Acids

Techniques for enhancing the tolerance of industrial microbes to abiotic stresses: A review

A novel point mutation in RpoB improves osmotolerance and succinic acid production in Escherichia coli

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