Role of Trehalose in Salinity and Temperature Tolerance in the Model Halophilic Bacterium Chromohalobacter salexigens

Reina-Bueno, Mercedes; Argandoña, Montserrat; Salvador, Manuel; Rodríguez-Moya, Javier; Iglesias‐Guerra, Fernando; Csonka, László N.; Nieto, Joaquı́n J.; Vargas, Carmen Regla

doi:10.1371/journal.pone.0033587

Cited by 58 publications

(31 citation statements)

References 73 publications

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“…Probably, trehalose adds stringency by shielding attractive forces between particles in a high salt concentration environment and maintaining enough repulsion to keep the particles apart. In plants and bacteria, trehalose has been reported to confer tolerance to high salt stress414243.…”

Section: Discussionmentioning

confidence: 99%

Trehalose prevents aggregation of exosomes and cryodamage

Bosch¹,

Beaurepaire²,

Allard³

et al. 2016

Sci Rep

242

206

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Exosomes are important mediators in intercellular communication. Released by many cell types, they transport proteins, lipids, and nucleic acids to distant recipient cells and contribute to important physiopathological processes. Standard current exosome isolation methods based on differential centrifugation protocols tend to induce aggregation of particles in highly concentrated suspensions and freezing of exosomes can induce damage and inconsistent biological activity. Trehalose is a natural, non-toxic sugar widely used as a protein stabilizer and cryoprotectant by the food and drug industry. Here we report that addition of 25 mM trehalose to pancreatic beta-cell exosome-like vesicle isolation and storage buffer narrows the particle size distribution and increases the number of individual particles per microgram of protein. Repeated freeze-thaw cycles induce an increase in particle concentration and in the width of the size distribution for exosome-like vesicles stored in PBS, but not in PBS 25 mM trehalose. No signs of lysis or incomplete vesicles were observed by cryo-electron tomography in PBS and trehalose samples. In macrophage immune assays, beta-cell extracellular vesicles in trehalose show consistently higher TNF-alpha cytokine secretion stimulation indexes suggesting improved preservation of biological activity. The addition of trehalose might be an attractive means to standardize experiments in the field of exosome research and downstream applications.

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Section: Discussionmentioning

confidence: 99%

Trehalose prevents aggregation of exosomes and cryodamage

Bosch¹,

Beaurepaire²,

Allard³

et al. 2016

Sci Rep

242

206

View full text Add to dashboard Cite

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“…Introduction of pHYDROX2 in wild-type cells switched the synthesis to hydroxyectoine, which became the major compatible solute, followed by glutamate, trehalose, and minor amounts of glucosylglycerol and ectoine. The presence of trehalose in the wild type overexpressing hydroxyectoine at 37°C was unexpected, as this sugar is synthesized by C. salexigens in response to heat stress or when ectoine is absent (18). This apparent induction of trehalose synthesis by hydroxyectoine will be investigated in a further work.…”

mentioning

confidence: 91%

Temperature- and Salinity-Decoupled Overproduction of Hydroxyectoine by Chromohalobacter salexigens

Rodríguez-Moya

Argandoña

Iglesias‐Guerra

et al. 2013

Appl Environ Microbiol

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Hydroxyectoine overproduction by the natural producer Chromohalobacter salexigens is presented in this study. Genetically engineered strains were constructed that at low salinity coexpressed, in a vector derived from a native plasmid, the ectoine (ectABC) and hydroxyectoine (ectD) genes under the control of the ectA promoter, in a temperature-independent manner. Hydroxyectoine production was further improved by increasing the copies of ectD and using a C. salexigens genetic background unable to synthesize ectoines.

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“…Denaturation of cytoplasmic proteins is another common consequence of these stresses (56,58). Trehalose, which facilitates protein folding and prevents aggregation of denatured proteins (59,60), is widely recognized as a thermoprotectant (61)(62)(63). Indeed, we showed that the missense mutation in otsA, encoding the key enzyme for trehalose synthesis (64,65), partly accounts for the improved thermotolerance of the evolved strains.…”

Section: Discussionmentioning

confidence: 83%

Thermal and Solvent Stress Cross-Tolerance Conferred to Corynebacterium glutamicum by Adaptive Laboratory Evolution

Oide

Gunji

Moteki

et al. 2015

Appl Environ Microbiol

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bReinforcing microbial thermotolerance is a strategy to enable fermentation with flexible temperature settings and thereby to save cooling costs. Here, we report on adaptive laboratory evolution (ALE) of the amino acid-producing bacterium Corynebacterium glutamicum under thermal stress. After 65 days of serial passage of the transgenic strain GLY3, in which the glycolytic pathway is optimized for alanine production under oxygen deprivation, three strains adapted to supraoptimal temperatures were isolated, and all the mutations they acquired were identified by whole-genome resequencing. Of the 21 mutations common to the three strains, one large deletion and two missense mutations were found to promote growth of the parental strain under thermal stress. Additive effects on thermotolerance were observed among these mutations, and the combination of the deletion with the missense mutation on otsA, encoding a trehalose-6-phosphate synthase, allowed the parental strain to overcome the upper limit of growth temperature. Surprisingly, the three evolved strains acquired cross-tolerance for isobutanol, which turned out to be partly attributable to the genomic deletion associated with the enhanced thermotolerance. The deletion involved loss of two transgenes, pfk and pyk, encoding the glycolytic enzymes, in addition to six native genes, and elimination of the transgenes, but not the native genes, was shown to account for the positive effects on thermal and solvent stress tolerance, implying a link between energy-producing metabolism and bacterial stress tolerance. Overall, the present study provides evidence that ALE can be a powerful tool to refine the phenotype of C. glutamicum and to investigate the molecular bases of stress tolerance.A high-GϩC, Gram-positive bacterium, Corynebacterium glutamicum, was first discovered in Japan as a natural producer of glutamic acid (1) and has been exploited for industrial production of amino acids for over 50 years. Microbes employed in industrial-scale fermentation encounter a variety of stresses (2, 3), including thermal stress, which stems from heat generated by metabolic activities of microbial catalysts. Careful control of temperature during fermentation processes is crucial to protect microbes from thermal stress and to achieve optimal productivity. Engineering microbes with improved tolerance for thermal stress enables fermentation with more flexible temperature requirements and thereby leads to reductions in cooling costs. Different approaches have been developed to improve bacterial stress tolerance (4-6). Among them, adaptive laboratory evolution (ALE) makes use of naturally occurring mutations to generate individuals better adapted to certain environments (7-9). ALE has been successfully applied to metabolic engineering, as well as evolutionary studies on different organisms (10-15). Studies on Escherichia coli provide evidence that ALE can be a powerful tool to ameliorate bacterial stress tolerance (16)(17)(18)(19)(20). Independent groups have reported on ALE of the bacter...

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Role of Trehalose in Salinity and Temperature Tolerance in the Model Halophilic Bacterium Chromohalobacter salexigens

Cited by 58 publications

References 73 publications

Trehalose prevents aggregation of exosomes and cryodamage

Trehalose prevents aggregation of exosomes and cryodamage

Temperature- and Salinity-Decoupled Overproduction of Hydroxyectoine by Chromohalobacter salexigens

Thermal and Solvent Stress Cross-Tolerance Conferred to Corynebacterium glutamicum by Adaptive Laboratory Evolution

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