scholarly journals Commitment to sporulation in Bacillus subtilis and its relationship to development of actinomycin resistance

1969 ◽  
Vol 113 (1) ◽  
pp. 29-37 ◽  
Author(s):  
Janet M. Sterlini ◽  
J. Mandelstam
Keyword(s):  
Alkaline Phosphatase ◽  
Bacillus Subtilis ◽  
Heat Resistance ◽  
Messenger Rna ◽  
Actinomycin D ◽  
Dipicolinic Acid ◽  
Vegetative State ◽  
Single Point

1. Experiments to determine the point of commitment to sporulation were carried out by restoring nutrients at different times to suspensions of sporulating Bacillus subtilis. 2. No single point of commitment to the process as a whole was found. Instead, the cells became committed in turn to the following successive events connected with sporulation: formation of alkaline phosphatase, development of refractility, synthesis of dipicolinic acid and development of heat-resistance. 3. Each point of commitment was followed within about 30min. by a period in which the event concerned ceased to be inhibited by actinomycin D. 4. The implication of these results is that each point of commitment is probably due to the formation of a species of long-lived messenger RNA and that, in any case, sporulation is regulated at the level of both transcription and translation. 5. It is also shown that sporulation and growth are perhaps not mutually exclusive functions and that histidase, an enzyme typical of the vegetative state, can be induced in sporulating suspensions.

scholarly journals Molecular Physiological Characterization of a High Heat Resistant Spore Forming Bacillus subtilis Food Isolate

2021 ◽  
Vol 9 (3) ◽  
pp. 667
Author(s):  
Zhiwei Tu ◽  
Peter Setlow ◽  
Stanley Brul ◽  
Gertjan Kramer
Keyword(s):  
Bacillus Subtilis ◽  
Heat Resistance ◽  
Dipicolinic Acid ◽  
Laboratory Strain ◽  
High Heat ◽  
High Heat Resistance ◽  
Vegetative Cells ◽  
Heat Resistant ◽  
Food Isolate ◽  
Wet Heat

Bacterial endospores (spores) are among the most resistant living forms on earth. Spores of Bacillus subtilis A163 show extremely high resistance to wet heat compared to spores of laboratory strains. In this study, we found that spores of B. subtilis A163 were indeed very wet heat resistant and released dipicolinic acid (DPA) very slowly during heat treatment. We also determined the proteome of vegetative cells and spores of B. subtilis A163 and the differences in these proteomes from those of the laboratory strain PY79, spores of which are much less heat resistant. This proteomic characterization identified 2011 proteins in spores and 1901 proteins in vegetative cells of B. subtilis A163. Surprisingly, spore morphogenic protein SpoVM had no homologs in B. subtilis A163. Comparing protein expression between these two strains uncovered 108 proteins that were differentially present in spores and 93 proteins differentially present in cells. In addition, five of the seven proteins on an operon in strain A163, which is thought to be primarily responsible for this strain’s spores high heat resistance, were also identified. These findings reveal proteomic differences of the two strains exhibiting different resistance to heat and form a basis for further mechanistic analysis of the high heat resistance of B. subtilis A163 spores.

scholarly journals The Products of the spoVA Operon Are Involved in Dipicolinic Acid Uptake into Developing Spores of Bacillus subtilis

2002 ◽  
Vol 184 (2) ◽  
pp. 584-587 ◽  
Author(s):  
Federico Tovar-Rojo ◽  
Monica Chander ◽  
Barbara Setlow ◽  
Peter Setlow
Keyword(s):  
Bacillus Subtilis ◽  
Heat Resistance ◽  
Dipicolinic Acid ◽  
Lytic Enzyme ◽  
Acid Uptake ◽  
Wild Type ◽  
Wet Heat

ABSTRACT Bacillus subtilis cells with mutations in the spoVA operon do not complete sporulation. However, a spoVA strain with mutations that remove all three of the spore’s functional nutrient germinant receptors (termed the ger3 mutations) or the cortex lytic enzyme SleB (but not CwlJ) did complete sporulation. ger3 spoVA and sleB spoVA spores lack dipicolinic acid (DPA) and have lower core wet densities and levels of wet heat resistance than wild-type or ger3 spores. These properties of ger3 spoVA and sleB spoVA spores are identical to those of ger3 spoVF and sleB spoVF spores that lack DPA due to deletion of the spoVF operon coding for DPA synthetase. Sporulation in the presence of exogenous DPA restored DPA levels in ger3 spoVF spores to 53% of the wild-type spore levels, but there was no incorporation of exogenous DPA into ger3 spoVA spores. These data indicate that one or more products of the spoVA operon are involved in DPA transport into the developing forespore during sporulation.

scholarly journals Maturation of Released Spores Is Necessary for Acquisition of Full Spore Heat Resistance during Bacillus subtilis Sporulation

2011 ◽  
Vol 77 (19) ◽  
pp. 6746-6754 ◽  
Author(s):  
Jose-Luis Sanchez-Salas ◽  
Barbara Setlow ◽  
Pengfei Zhang ◽  
Yong-qing Li ◽  
Peter Setlow
Keyword(s):  
Bacillus Subtilis ◽  
Heat Resistance ◽  
Uv Radiation ◽  
Dipicolinic Acid ◽  
Sporulation Medium ◽  
Content Type ◽  
Dry Heat ◽  
Lower Resistance ◽  
Wet Heat ◽  
Spore Maturation

ABSTRACTThe first ∼10% of spores released from sporangia (early spores) duringBacillus subtilissporulation were isolated, and their properties were compared to those of the total spores produced from the same culture. The early spores had significantly lower resistance to wet heat and hypochlorite than the total spores but identical resistance to dry heat and UV radiation. Early and total spores also had the same levels of core water, dipicolinic acid, and Ca and germinated similarly with several nutrient germinants. The wet heat resistance of the early spores could be increased to that of total spores if early spores were incubated in conditioned sporulation medium for ∼24 h at 37°C (maturation), and some hypochlorite resistance was also restored. The maturation of early spores took place in pH 8 buffer with Ca2+but was blocked by EDTA; maturation was also seen with early spores of strains lacking the CotE protein or the coat-associated transglutaminase, both of which are needed for normal coat structure. Nonetheless, it appears to be most likely that it is changes in coat structure that are responsible for the increased resistance to wet heat and hypochlorite upon early spore maturation.

scholarly journals Assessment of Heat Resistance of Bacterial Spores from Food Product Isolates by Fluorescence Monitoring of Dipicolinic Acid Release

2005 ◽  
Vol 71 (7) ◽  
pp. 3556-3564 ◽  
Author(s):  
Remco Kort ◽  
Andrea C. O'Brien ◽  
Ivo H. M. van Stokkum ◽  
Suus J. C. M. Oomes ◽  
Wim Crielaard ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Heat Resistance ◽  
Temperature Dependence ◽  
Incubation Time ◽  
Dipicolinic Acid ◽  
Food Product ◽  
Microtiter Plate ◽  
Quantitative Model ◽  
Food Samples ◽  
Heat Inactivation

ABSTRACT This study is aimed at the development and application of a convenient and rapid optical assay to monitor the wet-heat resistance of bacterial endospores occurring in food samples. We tested the feasibility of measuring the release of the abundant spore component dipicolinic acid (DPA) as a probe for heat inactivation. Spores were isolated from the laboratory type strain Bacillus subtilis 168 and from two food product isolates, Bacillus subtilis A163 and Bacillus sporothermodurans IC4. Spores from the lab strain appeared much less heat resistant than those from the two food product isolates. The decimal reduction times (D values) for spores from strains 168, A163, and IC4 recovered on Trypticase soy agar were 1.4, 0.7, and 0.3 min at 105°C, 120°C, and 131°C, respectively. The estimated Z values were 6.3°C, 6.1°C, and 9.7°C, respectively. The extent of DPA release from the three spore crops was monitored as a function of incubation time and temperature. DPA concentrations were determined by measuring the emission at 545 nm of the fluorescent terbium-DPA complex in a microtiter plate fluorometer. We defined spore heat resistance as the critical DPA release temperature (Tc ), the temperature at which half the DPA content has been released within a fixed incubation time. We found Tc values for spores from Bacillus strains 168, A163, and IC4 of 108°C, 121°C, and 131°C, respectively. On the basis of these observations, we developed a quantitative model that describes the time and temperature dependence of the experimentally determined extent of DPA release and spore inactivation. The model predicts a DPA release rate profile for each inactivated spore. In addition, it uncovers remarkable differences in the values for the temperature dependence parameters for the rate of spore inactivation, DPA release duration, and DPA release delay.

scholarly journals A Two-Dimensional Protein Gel Electrophoresis Study of the Heat Stress Response of Bacillus subtilis Cells during Sporulation

2000 ◽  
Vol 182 (17) ◽  
pp. 4758-4763 ◽  
Author(s):  
Sara Movahedi ◽  
William Waites
Keyword(s):  
Bacillus Subtilis ◽  
Heat Shock ◽  
Heat Resistance ◽  
Gel Electrophoresis ◽  
Stress Proteins ◽  
Dipicolinic Acid ◽  
Two Dimensional ◽  
Maximum Increase ◽  
Two Dimensional Gel Electrophoresis ◽  
Heat Stress Response

ABSTRACT The heat resistance of spores of Bacillus subtilisformed at 30°C was enhanced by pretreatment at 48°C for 30 min, 60 min into sporulation, for all four strains examined. High-resolution two-dimensional gel electrophoresis showed the generation and/or overexpression of 60 proteins, 11 of which were specific to heat shock, concurrent to this acquired thermotolerance. The greatest number of new proteins was observed between 30 and 60 min after heat shock, and the longer the time between exponential growth and heat treatment, the fewer differences were observed on corresponding protein profiles. The time at which heating produced the maximum increase in spore resistance and the most new proteins on two-dimensional gels occurred before alkaline phosphatase and dipicolinic acid production and corresponded to stage I or II of sporulation. The stress proteins formed disappeared later in sporulation, suggesting that heat shock proteins increase spore heat resistance by altering spore structure rather than by repairing heat damage during germination and outgrowth.

scholarly journals Sporulation in Bacillus subtilis. Morphological changes

1968 ◽  
Vol 109 (5) ◽  
pp. 819-824 ◽  
Author(s):  
D. Kay ◽  
S. C. Warren
Keyword(s):  
Bacillus Subtilis ◽  
Heat Resistance ◽  
Early Stage ◽  
Morphological Changes ◽  
Dipicolinic Acid ◽  
Close Contact ◽  
Spore Coat ◽  
Serial Sections ◽  
Filament Formation ◽  
Thin Sections

1. When Bacillus subtilis was grown in a medium in which sporulation occurred well-defined morphological changes were seen in thin sections of the cells. 2. Over a period of 7·5hr. beginning 2hr. after the initiation of sporulation the following major stages were observed: axial nuclear-filament formation, spore-septum formation, release of the fore-spore within the cell, development of the cortex around the fore-spore, the laying down of the spore coat and the completion of the corrugated spore coat before release of the spore from the mother cell. 3. The appearance of refractile bodies and 2,6-dipicolinic acid and the development of heat-resistance began between 5 and 6·5hr. after initiation of sporulation. 4. The appearance of 2,6-dipicolinic acid and the onset of refractility appeared to coincide with a diminution of electron density in the spore core and cortex. 5. Heat-resistance was associated with the terminal stage, the completion of the spore coat. 6. The spore coat was composed of an inner and an outer layer, each of which consisted of three or four electron-dense laminae. 7. Serial sections through cells at an early stage of sporulation showed that the membranes of each spore septum were always continuous with the membranes of a mesosome, which was itself in close contact with the bacterial or spore nucleoid. 8. These changes were correlated with biochemical events occurring during sporulation.

Properties of spores of Bacillus subtilis with or without a transposon that decreases spore germination and increases spore wet heat resistance

2021 ◽  
Author(s):  
Yannong Luo ◽  
George Korza ◽  
Angela M. DeMarco ◽  
Oscar P. Kuipers ◽  
Yong‐qing Li ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Heat Resistance ◽  
Spore Germination ◽  
Wet Heat

scholarly journals Role of SpoVA Proteins in Release of Dipicolinic Acid during Germination of Bacillus subtilis Spores Triggered by Dodecylamine or Lysozyme

2006 ◽  
Vol 189 (5) ◽  
pp. 1565-1572 ◽  
Author(s):  
Venkata Ramana Vepachedu ◽  
Peter Setlow
Keyword(s):  
Bacillus Subtilis ◽  
Small Molecules ◽  
Spore Germination ◽  
Dipicolinic Acid ◽  
Temperature Sensitive ◽  
Wild Type ◽  
Ph Optimum ◽  
Inner Membrane ◽  
Temperature Sensitive Mutants

ABSTRACT The release of dipicolinic acid (DPA) during the germination of Bacillus subtilis spores by the cationic surfactant dodecylamine exhibited a pH optimum of ∼9 and a temperature optimum of 60°C. DPA release during dodecylamine germination of B. subtilis spores with fourfold-elevated levels of the SpoVA proteins that have been suggested to be involved in the release of DPA during nutrient germination was about fourfold faster than DPA release during dodecylamine germination of wild-type spores and was inhibited by HgCl2. Spores carrying temperature-sensitive mutants in the spoVA operon were also temperature sensitive in DPA release during dodecylamine germination as well as in lysozyme germination of decoated spores. In addition to DPA, dodecylamine triggered the release of amounts of Ca2+ almost equivalent to those of DPA, and at least one other abundant spore small molecule, glutamic acid, was released in parallel with Ca2+ and DPA. These data indicate that (i) dodecylamine triggers spore germination by opening a channel in the inner membrane for Ca2+-DPA and other small molecules, (ii) this channel is composed at least in part of proteins, and (iii) SpoVA proteins are involved in the release of Ca2+-DPA and other small molecules during spore germination, perhaps by being a part of a channel in the spore's inner membrane.

Effects of Cold Atmospheric Pressure Plasma Jeton the Viability of Bacillus subtilis Endospores

2013 ◽  
Vol 647 ◽  
pp. 524-531
Author(s):  
Vinita Sharma ◽  
Katsuhiko Hosoi ◽  
Tamio Mori ◽  
Shin-ichi Kuroda
Keyword(s):  
Bacillus Subtilis ◽  
Atmospheric Pressure ◽  
Dipicolinic Acid ◽  
Atmospheric Pressure Plasma ◽  
Dilution Method ◽  
Pressure Plasma ◽  
Agar Disc ◽  
Bacterial Endospores ◽  
Cold Atmospheric Pressure Plasma ◽  
Ar Gas

In this study, we conducted experiments to investigate the effectiveness of a non-equilibrium Ar-N2 plasma jet generated by a Cold Atmospheric Pressure Plasma Torch (CAPPLAT) at a sinusoidal voltage of 20 kV, frequency of 30 kHz with 10 slm of Ar gas and 100 sccm of N2 gas. Highly environmental stress resistant bacterial endospores of Bacillus subtilis, dried on an agar disc were exposed to the plasma discharge from the CAPPLAT for different durations. The viability of spores after plasma exposure was checked by counting CFUs by serial dilution method. We also measured the amount of released DPA (dipicolinic acid, pyridine-2, 6-dicarboxylic acid), which is exclusively found in endospore protoplast (cortex), to confirm the disintegration of the cortex. We could successfully inactivate a population of Bacillus endospores of about 1.0 × 107 to 4.0 × 107 spores/ml.

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