Role of Dipicolinic Acid in the Germination, Stability, and Viability of Spores of
            <i>Bacillus subtilis</i>

Magge, Anil; Granger, Amanda C.; Wahome, Paul G.; Setlow, Barbara; Vepachedu, Venkata Ramana; Loshon, Charles A.; Peng, Lixin; Chen, De; Li, Yongqing; Setlow, Peter

doi:10.1128/jb.00477-08

Cited by 82 publications

(81 citation statements)

References 37 publications

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“…The one exception to this first conclusion is that ⌬T release increases significantly in spores of B. megaterium and B. subtilis that lack the CLE CwlJ (26,27). Presumably this is because the initial slow release of DPA during germination normally triggers rapid CwlJ action that in turn increases the rate of further DPA release, while this is not how SleB is activated during germination (20,29).…”

Section: Discussionmentioning

confidence: 78%

“…Ca-DPA release then results in the activation of two redundant cortex-lytic enzymes (CLEs), CwlJ and SleB, which hydrolyze the spore's peptidoglycan cortex layer (16,22,27,29). CwlJ is activated by Ca-DPA as it is released from the spore while SleB is activated only after most DPA is released (17,20,22,26,27). Cortex hydrolysis ultimately allows the spore core to expand and take up more water, raising the core water content from the 35 to 45% of wet weight in the dormant spore to the 80% of wet weight characteristic of growing cells.…”

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confidence: 99%

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Factors Affecting Variability in Time between Addition of Nutrient Germinants and Rapid Dipicolinic Acid Release during Germination of Spores of Bacillus Species

Zhang

Garner²,

Yi³

et al. 2010

J Bacteriol

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105

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The simultaneous nutrient germination of hundreds of individual wild-type spores of three Bacillus species and a number of Bacillus subtilis strains has been measured by two new methods, and rates of release of the great majority of the large pool of dipicolinic acid (DPA) from individual spores of B. subtilis strains has been measured by Raman spectroscopy with laser tweezers. The results from these analyses and published data have allowed a number of significant conclusions about the germination of spores of Bacillus species as follows. (i) The time needed for release of the great majority of a Bacillus spore's DPA once rapid DPA release had begun (⌬T release ) during nutrient germination was independent of the concentration of nutrient germinant used, the level of the germinant receptors (GRs) that recognize nutrient germinants used and heat activation prior to germination. Values for ⌬T release were generally 0.5 to 3 min at 25 to 37°C for individual wild-type spores. (ii) Despite the conclusion above, germination of individual spores in populations was very heterogeneous, with some spores in wild-type populations completing germination >15-fold slower than others. (iii) The major factor in the heterogeneity in germination of individual spores in populations was the highly variable lag time, T lag , between mixing spores with nutrient germinants and the beginning of ⌬T release . (iv) A number of factors decrease spores' T lag values including heat activation, increased levels of GRs/spore, and higher levels of nutrient germinants. These latter factors appear to affect the level of activated GRs/spore during nutrient germination. (v) The conclusions above lead to the simple prediction that a major factor causing heterogeneity in Bacillus spore germination is the number of functional GRs in individual spores, a number that presumably varies significantly between spores in populations.Spores of various Bacillus species are metabolically dormant and can survive for years in this state (30). However, spores constantly sense their environment, and if appropriate small molecules termed germinants are present, spores can rapidly return to life in the process of germination followed by outgrowth (25,29,30). The germinants that most likely trigger spore germination in the environment are low-molecularweight nutrient molecules, the identities of which are strain and species specific, including amino acids, sugars, and purine nucleosides. Metabolism of these nutrient germinants is not needed for the triggering of spore germination. Rather, these germinants are recognized by germinant receptors (GRs) located in the spore's inner membrane that recognize their cognate germinants in a stereospecific manner (17,24,25,29). Spores have a number of such GRs, with three functional GRs in Bacillus subtilis spores and even more in Bacillus anthracis, Bacillus cereus, and Bacillus megaterium spores (6,29,30).Binding of nutrient germinants to some single GRs is sufficient to trigger spore germination, for example the triggering of B...

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Section: Discussionmentioning

confidence: 78%

mentioning

confidence: 99%

Factors Affecting Variability in Time between Addition of Nutrient Germinants and Rapid Dipicolinic Acid Release during Germination of Spores of Bacillus Species

Zhang

Garner²,

Yi³

et al. 2010

J Bacteriol

Self Cite

105

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“…Spores of B. subtilis strains that cannot synthesize DPA are also unstable and lyse during sporulation (13,40). B. subtilis spores with significantly lower than normal DPA levels can be isolated, but they exhibit relatively rapid spontaneous germination (28). Thus, B. subtilis spores with low or no DPA levels, for whatever reason, spontaneously and rapidly trigger subsequent events in the spore germination process, most notably the hydrolysis of cortex PG.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Clostridium perfringens Spores That Lack SpoVA Proteins and Dipicolinic Acid

Paredes-Sabja¹,

Setlow

et al. 2008

J Bacteriol

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Spores of Clostridium perfringens possess high heat resistance, and when these spores germinate and return to active growth, they can cause gastrointestinal disease. Work with Bacillus subtilis has shown that the spore's dipicolinic acid (DPA) level can markedly influence both spore germination and resistance and that the proteins encoded by the spoVA operon are essential for DPA uptake by the developing spore during sporulation. We now find that proteins encoded by the spoVA operon are also essential for the uptake of Ca 2؉ and DPA into the developing spore during C. perfringens sporulation. Spores of a spoVA mutant had little, if any, Ca 2؉ and DPA, and their core water content was approximately twofold higher than that of wild-type spores. These DPA-less spores did not germinate spontaneously, as DPA-less B. subtilis spores do. Indeed, wild-type and spoVA C. perfringens spores germinated similarly with a mixture of L-asparagine and KCl (AK), KCl alone, or a 1:1 chelate of Ca 2؉ and DPA (Ca-DPA). However, the viability of C. perfringens spoVA spores was 20-fold lower than the viability of wild-type spores. Decoated wild-type and spoVA spores exhibited little, if any, germination with AK, KCl, or exogenous Ca-DPA, and their colony-forming efficiency was 10 3 -to 10 4 -fold lower than that of intact spores. However, lysozyme treatment rescued these decoated spores. Although the levels of DNAprotective ␣/␤-type, small, acid-soluble spore proteins in spoVA spores were similar to those in wild-type spores, spoVA spores exhibited markedly lower resistance to moist heat, formaldehyde, HCl, hydrogen peroxide, nitrous acid, and UV radiation than wild-type spores did. In sum, these results suggest the following. (i) SpoVA proteins are essential for Ca-DPA uptake by developing spores during C. perfringens sporulation. (ii) SpoVA proteins and Ca-DPA release are not required for C. perfringens spore germination. (iii) A low spore core water content is essential for full resistance of C. perfringens spores to moist heat, UV radiation, and chemicals.Clostridium perfringens food poisoning is caused mainly by enterotoxigenic type A isolates that have the ability to form metabolically dormant spores that are extremely resistant to heat, radiation, and toxic chemicals (45,49,50,55). These highly resistant spores can survive traditional methods for cooking meat and poultry products as well as other processing treatments used in the food industry. The surviving spores can then go through germination and outgrowth, generating the vegetative cells that cause disease (30).Spores of Bacillus species also have the extreme resistance of C. perfringens spores. The factors involved in Bacillus subtilis spore resistance include the following: (i) relatively impermeable spore inner membrane; (ii) low water content of the spore core; (iii) high levels of pyridine-2,6-dicarboxylic acid (dipicolinic acid [DPA]) in the spore core and the type and amount of cations chelated by DPA; and (iv) the saturation of spore DNA with ␣/␤-type small, acid-s...

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“…Other than participating in energy metabolism and amino acid biosynthesis (particularly the BCAAs), during sporulation, pyruvate is also used for high-level synthesis of dipicolinic acid (ϳ25% of sporal core dry weight), which is important for spore germination and resistance (56). Consequently, pyruvate produced merely from the EMP pathway would be insufficient during sporulation.…”

Section: Fig 2 Cog Analysismentioning

confidence: 99%

The Metabolic Regulation of Sporulation and Parasporal Crystal Formation in Bacillus thuringiensis Revealed by Transcriptomics and Proteomics

Wang

Han

Cao

et al. 2013

Molecular & Cellular Proteomics

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Bacillus thuringiensis is a well-known entomopathogenic bacterium used worldwide as an environmentally compatible biopesticide. During sporulation, B. thuringiensis accumulates a large number of parasporal crystals consisting of insecticidal crystal proteins (ICPs) that can account for nearly 20 -30% of the cell's dry weight. However, the metabolic regulation mechanisms of ICP synthesis remain to be elucidated. In this study, the combined efforts in transcriptomics and proteomics mainly uncovered the following 6 metabolic regulation mechanisms: (1) proteases and the amino acid metabolism (particularly, the branched-chain amino acids) became more active during sporulation; (2) stored poly-␤-hydroxybutyrate and acetoin, together with some low-quality substances provided considerable carbon and energy sources for sporulation and parasporal crystal formation; (3) the pentose phosphate shunt demonstrated an interesting regulation mechanism involving gluconate when CT-43 cells were grown in GYS medium; (4) the tricarboxylic acid cycle was significantly modified during sporulation; (5) an obvious increase in the quantitative levels of enzymes and cytochromes involved in energy production via the electron transport system was observed; (6) most F 0 F 1 -ATPase subunits were remarkably up-regulated during sporulation. This study, for the first time, systematically reveals the metabolic regulation mechanisms involved in the supply of amino acids, carbon substances, and energy for B. thuringiensis spore and parasporal crystal formation at both the transcriptional and translational levels. Molecular & Cellular Proteomics

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Role of Dipicolinic Acid in the Germination, Stability, and Viability of Spores of Bacillus subtilis

Cited by 82 publications

References 37 publications

Factors Affecting Variability in Time between Addition of Nutrient Germinants and Rapid Dipicolinic Acid Release during Germination of Spores of Bacillus Species

Factors Affecting Variability in Time between Addition of Nutrient Germinants and Rapid Dipicolinic Acid Release during Germination of Spores of Bacillus Species

Characterization of Clostridium perfringens Spores That Lack SpoVA Proteins and Dipicolinic Acid

The Metabolic Regulation of Sporulation and Parasporal Crystal Formation in Bacillus thuringiensis Revealed by Transcriptomics and Proteomics

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