Dependence of the Heat Resistance of Bacterial Endospores on their Dipicolinic Acid Content

Church, Brooks D.; Halvorson, Harlyn O.

doi:10.1038/183124a0

Cited by 141 publications

(72 citation statements)

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“…Ca-DPA levels in dormant Bacillus spores can vary with the species, strain, and sporulation conditions (7,11,13,18,22,27). These levels could also vary between individual spores in a spore population, perhaps due to cellular heterogeneity (2,8).…”

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Levels of Ca ²⁺ -Dipicolinic Acid in Individual Bacillus Spores Determined Using Microfluidic Raman Tweezers

et al. 2007

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Pyridine-2,6-dicarboxylic acid (dipicolinic acid [DPA]) in a 1:1 chelate with calcium ion (Ca-DPA) comprises 5 to 15% of the dry weight of spores of Bacillus species. Ca-DPA is important in spore resistance to many environmental stresses and in spore stability, and Ca-DPA levels in spore populations can vary with spore species/strains, as well as with sporulation conditions. We have measured levels of Ca-DPA in large numbers of individual spores in populations of a variety of Bacillus species and strains by using microfluidic Raman tweezers, in which a single spore is trapped in a focused laser beam and its Ca-DPA is quantitated from the intensity of the Ca-DPA-specific band at 1,017 cm ؊1 in Raman spectroscopy. Conclusions from these measurements include the following: (i) Ca-DPA concentrations in the spore core are >800 mM, well above Ca-DPA solubility; (ii) SpoVA proteins may be involved in Ca-DPA uptake in sporulation; and (iii) Ca-DPA levels differ significantly among individual spores in a population, but much of this variation could be due to variations in the sizes of individual spores.

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“…This process can elevate spore resistance to wet heat by protecting core proteins from inactivation or denaturation. Ca-DPA also functions to protect spore DNA against a variety of damaging agents, including dry heat, desiccation, UV radiation, and some chemicals (1,7,22,27,32,39).…”

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Levels of Ca ²⁺ -Dipicolinic Acid in Individual Bacillus Spores Determined Using Microfluidic Raman Tweezers

et al. 2007

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“…Mutants producing stable DPA-less spores of several Bacillus species were isolated more than 40 years ago (1, 10, 36), but these strains have not been characterized and may have had multiple mutations. B. cereus spores containing no DPA or low DPA levels have also been generated by addition of phenylalanine to or omission of particular nutrients or Ca 2ϩ from sporulation media (4,8,15). Notably, a significant percentage of the latter DPA-deficient B. cereus spores germinated spontaneously in the absence of nutrient germinants.…”

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

et al. 2008

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Spores of Bacillus subtilis spoVF strains that cannot synthesize dipicolinic acid (DPA) but take it up during sporulation were prepared in medium with various DPA concentrations, and the germination and viability of these spores as well as the DPA content in individual spores were measured. Levels of some other small molecules in DPA-less spores were also measured. These studies have allowed the following conclusions. (i) Spores with no DPA or low DPA levels that lack either the cortex-lytic enzyme (CLE) SleB or the receptors that respond to nutrient germinants could be isolated but were unstable and spontaneously initiated early steps in spore germination. (ii) Spores that lacked SleB and nutrient germinant receptors and also had low DPA levels were more stable.

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“…The presence of DPA in bacterial spores was first reported by Powell [3] in 1953. It has been suggested that it is involved in spore dormancy [4,5], spore stability [6], germination [7], heat resistance [4,5,[8][9][10] and ultraviolet (UV) and gamma irradiation resistancef 11,12]. DPA is primarily located in the spore core and has been proposed to be tightly packed with the vital cell constituents, linked with proteins or amino acids, or intercalated with DNA.…”

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Two-dimensional multiwavelength fluorescence spectra of dipicolinic acid and calcium dipicolinate

Sarasanandarajah¹,

Kunnil²,

Bronk

et al. 2005

Appl. Opt.

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Public reporting burden (or this collection of information is estimated AUTHOR(S)Sivananthan Sarasanandarajah, Joseph Kunnil, Burt V. Bronk, Lou Reinisch PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES)Department of Physics and Astronomy University of Canterbury Private Bag 4800 Christchurch, New Zealand SPONSORING/MONITORING AGENCY NAME(S) AND ADDRESS(ES)Air AFRL-HE-WP-TR-2004-0140 SUPPLEMENTARY NOTES 12a. DISTRIBUTION AVAILABILITY STATEMENTApproved for Public Release; distribution is unlimited. 12b DISTRIBUTION CODE ABSTRACT (Max/mum 200 words)Dipicolinic acid (DPA) and the Ca2+ complex of DPA (CaDPA) are major chemical components of bacterial spores. With fluorescence being considered for the detection and identification of spores, it is important to understand the optical properties of the major components of the spores. In this paper we report in some detail on the room temperature fluorescence excitation and emission spectra of DPA and its calcium ion complex and comparison of the excitation-emission spectrum in a dry, wet paste and aqueous form. DPA solutions have very weak, if any, fluorescence and it is only slightly greater in the dry state. After the exposure to a broad source UV light of the DPA, wet or dry, we observe a large increase in fluorescence with a maximum intensity emission peak at around 440 nm for excitation light with wavelength around 360 nm. There is a slight blue shift in the absorption spectra of UV exposed DPA from the unexposed DPA solution. CaDPA in solution and dried show very slight fluorescence and a substantial increase of fluorescence was observed after UV exposure with emission peak around 410 nm for excitation around 305 nm. The detailed excitation-emission spectra presented here are necessary for better interpretation of the fluorescence spectra of bacterial spores where DPA is a major chemical component.

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Dependence of the Heat Resistance of Bacterial Endospores on their Dipicolinic Acid Content

Cited by 141 publications

References 3 publications

Levels of Ca ²⁺ -Dipicolinic Acid in Individual Bacillus Spores Determined Using Microfluidic Raman Tweezers

Levels of Ca ²⁺ -Dipicolinic Acid in Individual Bacillus Spores Determined Using Microfluidic Raman Tweezers

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

Two-dimensional multiwavelength fluorescence spectra of dipicolinic acid and calcium dipicolinate

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Dependence of the Heat Resistance of Bacterial Endospores on their Dipicolinic Acid Content

Cited by 141 publications

References 3 publications

Levels of Ca 2+ -Dipicolinic Acid in Individual Bacillus Spores Determined Using Microfluidic Raman Tweezers

Levels of Ca 2+ -Dipicolinic Acid in Individual Bacillus Spores Determined Using Microfluidic Raman Tweezers

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

Two-dimensional multiwavelength fluorescence spectra of dipicolinic acid and calcium dipicolinate

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Levels of Ca ²⁺ -Dipicolinic Acid in Individual Bacillus Spores Determined Using Microfluidic Raman Tweezers

Levels of Ca ²⁺ -Dipicolinic Acid in Individual Bacillus Spores Determined Using Microfluidic Raman Tweezers