The Effects of Deuterium Oxide on Certain Microorganisms*

Giovanni, Rosalie De

doi:10.1111/j.1749-6632.1960.tb39097.x

Cited by 15 publications

(6 citation statements)

References 6 publications

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“…After the discovery of D in 1932, it was shown in numerous experiments that extreme (>1000-fold elevated) D concentrations have major, mainly toxic effects to all kingdoms of life (recently reviewed in reference [12]): bacteriophages [13], bacteria [14], yeast [15], algae [16], plants [17], Drosophila melanogaster [18] and mammalian cell lines [19] are, to various extents, impaired in growth or reproduction. When exposing mammals to D 2 O, a wide range of symptoms, including lymphopenia, hypoglycemia, myocardial abnormality, generalized muscular weakness and impaired reproductive capacity were observed [20,21].…”

Section: Introductionmentioning

confidence: 99%

Deuterium-depletion has no significant impact on the mutation rate of Escherichia coli, deuterium abundance therefore has a probabilistic, not deterministic effect on spontaneous mutagenesis

Ajibola

Karcagi

Somlyai³

et al. 2021

PLoS ONE

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Deuterium (D), the second most abundant isotope of hydrogen is present in natural waters at an approximate concentration of 145–155 ppm (ca. 1.5E-4 atom/atom). D is known to influence various biological processes due to its physical and chemical properties, which significantly differ from those of hydrogen. For example, increasing D-concentration to >1000-fold above its natural abundance has been shown to increase the frequency of genetic mutations in several species. An interesting deterministic hypothesis, formulated with the intent of explaining the mechanism of D-mutagenicity is based on the calculation that the theoretical probability of base pairs to comprise two adjacent D-bridges instead of H-bridges is 2.3E-8, which is equal to the mutation rate of certain species. To experimentally challenge this hypothesis, and to infer the mutagenicity of D present at natural concentrations, we investigated the effect of a nearly 100-fold reduction of D concentration on the bacterial mutation rate. Using fluctuation tests, we measured the mutation rate of three Escherichia coli genes (cycA, ackA and galK) in media containing D at either <2 ppm or 150 ppm concentrations. Out of 15 pair-wise fluctuation analyses, nine indicated a significant decrease, while three marked the significant increase of the mutation/culture value upon D-depletion. Overall, growth in D-depleted minimal medium led to a geometric mean of 0.663-fold (95% confidence interval: 0.483–0.911) change in the mutation rate. This falls nowhere near the expected 10,000-fold reduction, indicating that in our bacterial systems, the effect of D abundance on the formation of point mutations is not deterministic. In addition, the combined results did not display a statistically significant change in the mutation/culture value, the mutation rate or the mutant frequency upon D-depletion. The potential mutagenic effect of D present at natural concentrations on E. coli is therefore below the limit of detection using the indicated methods.

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Section: Introductionmentioning

confidence: 99%

Deuterium-depletion has no significant impact on the mutation rate of Escherichia coli, deuterium abundance therefore has a probabilistic, not deterministic effect on spontaneous mutagenesis

Ajibola

Karcagi

Somlyai³

et al. 2021

PLoS ONE

View full text Add to dashboard Cite

show abstract

Section: Introductionmentioning

confidence: 99%

“…After the discovery of D in 1932, it was shown in numerous experiments that extreme (>1000-fold elevated) D concentrations have major, mainly toxic effects to all kingdoms of life: bacteriophages [12], bacteria [13], yeast [14], algae [15], plants [16], Drosophila melanogaster [17] and mammalian cell lines [18] are all inhibited in growth or reproduction. When exposing mammals to D 2 O, a wide range of symptoms, including lymphopenia, hypoglycemia, myocardial abnormality, generalized muscular weakness and impaired reproductive capacity were observed [19, 20].…”

Section: Introductionmentioning

confidence: 99%

“…The mutagenic effect of deuterium has also been in the focus of multiple investigations for several species. The presence of high (50-99.8 %) D 2 O concentration was shown to elevate mutation frequencies in bacteriophage T4 [12], in various bacteria including Escherichia coli [13], Salmonella typhimurium [23], Proteus mirabilis [24] and Streptococcus pneumoniae [25], and in Oryza sativa [16]. The observable changes in mutation frequencies were often strain and locus-dependent, but on average meant an order of magnitude of increase.…”

Section: Introductionmentioning

confidence: 99%

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Deuterium-depletion has no significant impact on the mutation rate ofEscherichia coli, invalidating the Double D-Bridge Hypothesis

Ajibola

Karcagi

Somlyai³

et al. 2020

Preprint

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Deuterium (D), the second most abundant isotope of hydrogen is present in natural waters at an approximate concentration of 145-155 ppm (cca. 1.5E-4 atom/atom). D is known to influence various biological processes due to its physical and chemical properties, which significantly differ from those of hydrogen. For example, increasing D-concentration to >1000-fold above its natural abundance has been shown to increase the frequency of genetic mutations in several species. The Double D-Bridge Hypothesis, formulated with the intent of explaining the mechanism of D-mutagenicity is based on the calculation that the theoretical frequency of A:T base pairs that comprise two D-bridges instead of H-bridges is 2E-8, which is equal to the mutation rate of certain species. To experimentally challenge this hypothesis, and to infer the mutagenicity of D present at natural concentrations, we investigated the effect of a nearly 100-fold reduction of D concentration on the bacterial mutation rate. Using fluctuation tests, we measured the mutation rate of three Escherichia coli genes (cycA, ackA and galK) in media containing D at either <2 ppm or 150 ppm concentrations. Out of 15 pair-wise fluctuation analyses, nine indicated a significant decrease, while three marked the significant increase of the mutation/culture value upon D-depletion. Overall, growth in D-depleted minimal medium led to a geometric mean of 0.663-fold (95% confidence interval: 0.483-0.911) change in the mutation rate. This falls nowhere near the expected 10,000-fold reduction, indicating that the Double D-Bridge Hypothesis does not hold. In addition, the combined results did not display a statistically significant change in the mutation/culture value, the mutation rate or the mutant frequency upon D-depletion. The potential mutagenic effect of D present at natural concentrations on E. coli is therefore below the limit of detection using the indicated methods.

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“…There are several limitations to this procedure. E. coli generally grows much slower in deuterated medium and often do not reach the same cell density as in hydrogenated media and solvent (Giovanni 1960), while the success of expression is often protein specific. Complicating matters further, purified protein must often be destabilized or refolded in H 2 O to allow for backexchange of amide protons incorporated into fully folded structural elements (Englander 2006).…”

Section: Introductionmentioning

confidence: 99%

Improving yields of deuterated, methyl labeled protein by growing in H2O

et al. 2018

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Solution NMR continues to make strides in addressing protein systems of significant size and complexity. A fundamental requirement to fully exploit the N-H TROSY and C-H methyl TROSY effects is highly deuterated protein. Unfortunately, traditional overexpression in Escherichia coli (E. coli) during growth on media prepared in DO leads to many difficulties and limitations, such as cell toxicity, decreased yield, and the need to unfold or destabilize proteins for back exchange of amide protons. These issues are exacerbated for non-ideal systems such as membrane proteins. Expression of protein during growth in HO, with the addition of H-labeled amino acids derived from algal extract, can potentially avoid these issues. We demonstrate a novel fermentation methodology for high-density bacterial growth in HO M9 medium that allows for appropriate isotopic labeling and deuteration. Yields are significantly higher than those achieved in DO M9 for a variety of protein targets while still achieving 75-80% deuteration. Because the procedure does not require bulk DO or deuterated glucose, the cost per liter of growth medium is significantly decreased; taking into account improvements in yield, these savings can be quite dramatic. Triple-labeled protein is also efficiently produced including specific CH labeling of isoleucine, leucine, and valine using the traditional ILV precursors in combination with an ILV-depleted mix of H/N amino acids. These results are demonstrated for the membrane protein sensory rhodopsin II and the soluble proteins human aldoketoreductase AKR1c3, human ubiquitin, and bacterial flavodoxin. Limitations of the approach in the context of very large molecular weight proteins are illustrated using the bacterial Lac repressor transcription factor.

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The Effects of Deuterium Oxide on Certain Microorganisms*

Cited by 15 publications

References 6 publications

Deuterium-depletion has no significant impact on the mutation rate of Escherichia coli, deuterium abundance therefore has a probabilistic, not deterministic effect on spontaneous mutagenesis

Deuterium-depletion has no significant impact on the mutation rate of Escherichia coli, deuterium abundance therefore has a probabilistic, not deterministic effect on spontaneous mutagenesis

Deuterium-depletion has no significant impact on the mutation rate ofEscherichia coli, invalidating the Double D-Bridge Hypothesis

Improving yields of deuterated, methyl labeled protein by growing in H2O

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