Glycine betaine as a cryoprotectant for prokaryotes

Cleland, David; Krader, Paul; McCree, Coral; Tang, Jane; Emerson, David

doi:10.1016/j.mimet.2004.02.015

Cited by 85 publications

(51 citation statements)

References 12 publications

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“…Replicate, 1-mL aliquots of water collected for single cell analyses were cryopreserved with 6% glycine betaine (Sigma-Aldrich) and stored at −80°C or in liquid nitrogen (46). Single cell sorting, whole-genome amplification, real-time PCR screens, and PCR product sequence analyses were performed at the Bigelow Laboratory Single Cell Genomics Center (www.bigelow.org/scgc), as described by Stepanauskas and Sieracki (13) for SAGs MS024-2A, MS024-3C, MS190-1F, and MS220-5C and by Swan et al (15) and Martinez-Garcia et al (14) for the remaining SAGs.…”

Section: Methodsmentioning

confidence: 99%

Prevalent genome streamlining and latitudinal divergence of planktonic bacteria in the surface ocean

Swan

Tupper

Sczyrba

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

326

319

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Planktonic bacteria dominate surface ocean biomass and influence global biogeochemical processes, but remain poorly characterized owing to difficulties in cultivation. Using large-scale single cell genomics, we obtained insight into the genome content and biogeography of many bacterial lineages inhabiting the surface ocean. We found that, compared with existing cultures, natural bacterioplankton have smaller genomes, fewer gene duplications, and are depleted in guanine and cytosine, noncoding nucleotides, and genes encoding transcription, signal transduction, and noncytoplasmic proteins. These findings provide strong evidence that genome streamlining and oligotrophy are prevalent features among diverse, freeliving bacterioplankton, whereas existing laboratory cultures consist primarily of copiotrophs. The apparent ubiquity of metabolic specialization and mixotrophy, as predicted from single cell genomes, also may contribute to the difficulty in bacterioplankton cultivation. Using metagenome fragment recruitment against single cell genomes, we show that the global distribution of surface ocean bacterioplankton correlates with temperature and latitude and is not limited by dispersal at the time scales required for nucleotide substitution to exceed the current operational definition of bacterial species. Single cell genomes with highly similar small subunit rRNA gene sequences exhibited significant genomic and biogeographic variability, highlighting challenges in the interpretation of individual gene surveys and metagenome assemblies in environmental microbiology. Our study demonstrates the utility of single cell genomics for gaining an improved understanding of the composition and dynamics of natural microbial assemblages.comparative genomics | marine microbiology | microbial ecology | microbial microevolution | operational taxonomic unit

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Section: Methodsmentioning

confidence: 99%

Prevalent genome streamlining and latitudinal divergence of planktonic bacteria in the surface ocean

Swan

Tupper

Sczyrba

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

326

319

View full text Add to dashboard Cite

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“…Despite the significant reduction in viability of both strains at 10°C, the probiotic expressing BetL displayed a slower death rate and was recovered in higher numbers than the control strain. In addition to chill tolerance, compatible solutes have also been suggested to improve cryotolerance (6,25). Indeed, betL expression in L. salivarius UCC118 derivatives leads to a significant increase in viability following repeated freeze-thaw cycles during storage at Ϫ20°C and Ϫ70°C.…”

Section: Vol 72 2006 Betl Expression In Lactobacillus Salivarius 2173mentioning

confidence: 99%

“…Another important role of compatible solutes is to counteract the damaging effects of water loss due to drying (6,22,23). Freeze-and spray-drying, both commonly used practices for the preparation and storage of concentrated stocks of LAB, expose cells to the undesirable attenuating effects of extreme temperatures, dehydration, protein denaturation, and membrane damage (2,12,28,50).…”

Section: Vol 72 2006 Betl Expression In Lactobacillus Salivarius 2173mentioning

confidence: 99%

Heterologous Expression of BetL, a Betaine Uptake System, Enhances the Stress Tolerance ofLactobacillus salivariusUCC118

Sheehan

Sleator

Fitzgerald

et al. 2006

Appl Environ Microbiol

119

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Given the increasing commercial and clinical relevance of probiotic cultures, improving the technological robustness of what are often process-sensitive cultures is an important biological goal. The nisin-controlled expression system was used to direct the heterologous expression of the listerial betaine uptake system BetL in the probiotic strain Lactobacillus salivarius UCC118. Following nisin induction, strains expressing betL exhibited a significant increase in resistance to several stresses, including elevated osmo-, cryo-, baro-, and chill tolerance, as well as increased resistance to spray-and freeze-drying. The ability to confer additional stress tolerance on a probiotic culture may be an important step in delivering viable cultures for maximal efficacy.

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“…GB appears to be the most effective osmolyte in growing B. subtilis cells, and it can accumulate to Ն500 mM (12,27). In addition to providing osmoprotection to growing B. subtilis cells, GB can also provide significant thermal protection (12) and has been reported to provide desiccation resistance to bacteria of a number of genera (3,5).…”

mentioning

confidence: 99%

“…GB is one of the most common and effective osmolytes found in bacteria and has strong stabilizing effects on macromolecules (4,5,8). Indeed, elevated GB levels increase the tolerance of growing B. subtilis cells to elevated temperatures (12).…”

mentioning

confidence: 99%

Levels of Glycine Betaine in Growing Cells and Spores of Bacillus Species and Lack of Effect of Glycine Betaine on Dormant Spore Resistance

et al. 2006

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Bacteria of various Bacillus species are able to grow in media with very high osmotic strength in part due to the accumulation of low-molecular-weight osmolytes such as glycine betaine (GB). Cells of Bacillus species grown in rich and minimal media contained low levels of GB, but GB levels were 4-to 60-fold higher in cells grown in media with high salt. GB levels in Bacillus subtilis cells grown in minimal medium were increased ϳ7-fold by GB in the medium and 60-fold by GB plus high salt. GB was present in spores of Bacillus species prepared in media with or without high salt but at lower levels than in comparable growing cells. With spores prepared in media with high salt, GB levels were highest in B. subtilis spores and >20-fold lower in B. cereus and B. megaterium spores. Athough GB levels in B. subtilis spores were elevated 15-to 30-fold by GB plus high salt in sporulation media, GB levels did not affect spore resistance. GB levels were similar in wild-type B. subtilis spores and spores that lacked major small, acid-soluble spore proteins but were much lower in spores that lacked dipicolinic acid.

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Glycine betaine as a cryoprotectant for prokaryotes

Cited by 85 publications

References 12 publications

Prevalent genome streamlining and latitudinal divergence of planktonic bacteria in the surface ocean

Prevalent genome streamlining and latitudinal divergence of planktonic bacteria in the surface ocean

Heterologous Expression of BetL, a Betaine Uptake System, Enhances the Stress Tolerance ofLactobacillus salivariusUCC118

Levels of Glycine Betaine in Growing Cells and Spores of Bacillus Species and Lack of Effect of Glycine Betaine on Dormant Spore Resistance

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