Temperature–pH effect upon germination of bacterial spores

1976 ◽  
Vol 22 (2) ◽  
pp. 322-323 ◽  
Author(s):  
Yuzaburo Ishida ◽  
Takashi Ishido ◽  
Hajime Kadota
Keyword(s):  
Bacillus Subtilis ◽  
Spore Germination ◽  
Incubation Temperature ◽  
Ph Effect ◽  
Bacterial Spores ◽  
Optimum Ph ◽  
Different Temperatures

Using several kinds of criteria for the germination of bacterial spores, germination–pH curves were drawn for Bacillus subtilis spores observed at different temperatures. The experiments revealed that optimum pH for spore germination was markedly changed by changing the incubation temperature; the optimum pH for germination was 7.4 at 37 °C and 5.4 at 10 °C. A possible mechanism involved in this phenomenon is discussed.

scholarly journals The temperature of growth and sporulation modulates the efficiency of spore-display in Bacillus subtilis

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Claudia Petrillo ◽  
Stefany Castaldi ◽  
Mariamichela Lanzilli ◽  
Anella Saggese ◽  
Giuliana Donadio ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Spore Production ◽  
Bacterial Spores ◽  
Heterologous Proteins ◽  
Dna Coding ◽  
Gene Coding ◽  
Different Temperatures ◽  
Mucosal Vaccines ◽  
Heterologous Antigens ◽  
Selection Of

Abstract Background Bacterial spores displaying heterologous antigens or enzymes have long been proposed as mucosal vaccines, functionalized probiotics or biocatalysts. Two main strategies have been developed to display heterologous molecules on the surface of Bacillus subtilis spores: (i) a recombinant approach, based on the construction of a gene fusion between a gene coding for a coat protein (carrier) and DNA coding for the protein to be displayed, and (ii) a non-recombinant approach, based on the spontaneous and stable adsorption of heterologous molecules on the spore surface. Both systems have advantages and drawbacks and the selection of one or the other depends on the protein to be displayed and on the final use of the activated spore. It has been recently shown that B. subtilis builds structurally and functionally different spores when grown at different temperatures; based on this finding B. subtilis spores prepared at 25, 37 or 42 °C were compared for their efficiency in displaying various model proteins by either the recombinant or the non-recombinant approach. Results Immune- and fluorescence-based assays were used to analyze the display of several model proteins on spores prepared at 25, 37 or 42 °C. Recombinant spores displayed different amounts of the same fusion protein in response to the temperature of spore production. In spores simultaneously displaying two fusion proteins, each of them was differentially displayed at the various temperatures. The display by the non-recombinant approach was only modestly affected by the temperature of spore production, with spores prepared at 37 or 42 °C slightly more efficient than 25 °C spores in adsorbing at least some of the model proteins tested. Conclusion Our results indicate that the temperature of spore production allows control of the display of heterologous proteins on spores and, therefore, that the spore-display strategy can be optimized for the specific final use of the activated spores by selecting the display approach, the carrier protein and the temperature of spore production.

scholarly journals Arginine dephosphorylation propels spore germination in bacteria

2019 ◽  
Vol 116 (28) ◽  
pp. 14228-14237 ◽  
Author(s):  
Bing Zhou ◽  
Maja Semanjski ◽  
Natalie Orlovetskie ◽  
Saurabh Bhattacharya ◽  
Sima Alon ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Spore Germination ◽  
Developmental Process ◽  
Bacterial Spores ◽  
Biological Functions ◽  
Post Translational Modification ◽  
Phosphorylated Proteins ◽  
Σ Factor ◽  
Transcriptional Machinery

Bacterial spores can remain dormant for years but possess the remarkable ability to germinate, within minutes, once nutrients become available. However, it still remains elusive how such instant awakening of cellular machineries is achieved. UtilizingBacillus subtilisas a model, we show that YwlE arginine (Arg) phosphatase is crucial for spore germination. Accordingly, the absence of the Arg kinase McsB accelerated the process. Arg phosphoproteome of dormant spores uncovered a unique set of Arg-phosphorylated proteins involved in key biological functions, including translation and transcription. Consequently, we demonstrate that during germination, YwlE dephosphorylates an Arg site on the ribosome-associated chaperone Tig, enabling its association with the ribosome to reestablish translation. Moreover, we show that Arg dephosphorylation of the housekeeping σ factor A (SigA), mediated by YwlE, facilitates germination by activating the transcriptional machinery. Subsequently, we reveal that transcription is reinitiated at the onset of germination and its recommencement precedes that of translation. Thus, Arg dephosphorylation elicits the most critical stages of spore molecular resumption, placing this unusual post-translational modification as a major regulator of a developmental process in bacteria.

scholarly journals The temperature of growth and sporulation modulates the efficiency of spore-display in Bacillus subtilis

2020 ◽  
Author(s):  
Claudia Petrillo ◽  
Stefany Castaldi ◽  
Mariamichela Lanzilli ◽  
Anella Saggese ◽  
Giuliana Donadio ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Spore Production ◽  
Bacterial Spores ◽  
Heterologous Proteins ◽  
Dna Coding ◽  
Gene Coding ◽  
Different Temperatures ◽  
Mucosal Vaccines ◽  
Heterologous Antigens ◽  
Selection Of

Abstract Background: Bacterial spores displaying heterologous antigens or enzymes have long been proposed as mucosal vaccines, functionalized probiotics or biocatalysts. Two main strategies have been developed to display heterologous molecules on the surface of Bacillus subtilis spores: i) a recombinant approach, based on the construction of a gene fusion between a gene coding for a coat protein (carrier) and DNA coding for the protein to be displayed, and ii) a non-recombinant approach, based on the spontaneous and stable adsorption of heterologous molecules on the spore surface. Both systems have advantages and drawbacks and the selection of one or the other depends on the protein to be displayed and on the final use of the activated spore. It has been recently shown that B. subtilis builds structurally and functionally different spores when grown at different temperatures; based on this finding B. subtilis spores prepared at 25, 37 or 42°C were compared for their efficiency in displaying various model proteins by either the recombinant or the non-recombinant approach.Results: Immune- and fluorescence-based assays were used to analyze the display of several model proteins on spores prepared at 25, 37 or 42°C. Recombinant spores displayed different amounts of the same fusion protein in response to the temperature of spore production. In spores simultaneously displaying two fusion proteins, each of them was differentially displayed at the various temperatures. The display by the non-recombinant approach was only modestly affected by the temperature of spore production, with spores prepared at 37 or 42°C slightly more efficient than 25°C spores in adsorbing at least some of the model proteins tested. Conclusion: Our results indicate that the temperature of spore production allows control of the display of heterologous proteins on spores and, therefore, that the spore-display strategy can be optimized for the specific final use of the activated spores by selecting the display approach, the carrier protein and the temperature of spore production.

scholarly journals Memory of Germinant Stimuli in Bacterial Spores

mBio ◽  
2015 ◽  
Vol 6 (6) ◽  
Author(s):  
Shiwei Wang ◽  
James R. Faeder ◽  
Peter Setlow ◽  
Yong-qing Li
Keyword(s):  
Spore Germination ◽  
Incubation Temperature ◽  
Dipicolinic Acid ◽  
Metastable States ◽  
Bacterial Spores ◽  
Early Event ◽  
Content Type ◽  
Subsequent Exposure ◽  
Display Memory ◽  
Multiple Species

ABSTRACT Bacterial spores, despite being metabolically dormant, possess the remarkable capacity to detect nutrients and other molecules in their environment through a biochemical sensory apparatus that can trigger spore germination, allowing the return to vegetative growth within minutes of exposure of germinants. We demonstrate here that bacterial spores of multiple species retain memory of transient exposures to germinant stimuli that can result in altered responses to subsequent exposure. The magnitude and decay of these memory effects depend on the pulse duration as well as on the separation time, incubation temperature, and pH values between the pulses. Spores of Bacillus species germinate in response to nutrients that interact with germinant receptors (GRs) in the spore's inner membrane, with different nutrient types acting on different receptors. In our experiments, B. subtilis spores display memory when the first and second germinant pulses target different receptors, suggesting that some components of spore memory are downstream of GRs. Furthermore, nonnutrient germinants, which do not require GRs, exhibit memory either alone or in combination with nutrient germinants, and memory of nonnutrient stimulation is found to be more persistent than that induced by GR-dependent stimuli. Spores of B. cereus and Clostridium difficile also exhibit germination memory, suggesting that memory may be a general property of bacterial spores. These observations along with experiments involving strains with mutations in various germination proteins suggest a model in which memory is stored primarily in the metastable states of SpoVA proteins, which comprise a channel for release of dipicolinic acid, a major early event in spore germination. IMPORTANCE Cellular memory is defined as a sustained response to a transient environmental stimulus, and yet its generation and storage have not been described in bacterial spores. We demonstrate here that bacterial spores of multiple species retain memory of transient exposures to germinant stimuli that can result in altered responses to subsequent exposure. Memory was induced by activation of germinant receptors (GRs) or by GR-independent germinants and was accessed by both GR-dependent and GR-independent germinants. Analysis of effects on memory of exposure to GR-dependent and GR-independent germinants as well as in spores lacking various germination proteins suggests a model in which memory is stored primarily in metastable states of SpoVA proteins which comprise a channel for release of spore dipicolinic acid. Spore memory can also significantly reduce the concentration of nutrient germinants necessary to trigger germination, and this may be used to respond to low levels of nutrient germinants.

scholarly journals The Temperature of Growth and Sporulation Modulates the Efficiency of Spore-Display in Bacillus Subtilis

2020 ◽  
Author(s):  
Claudia Petrillo ◽  
Stefany Castaldi ◽  
Mariamichela Lanzilli ◽  
Anella Saggese ◽  
Giuliana Donadio ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Spore Production ◽  
Bacterial Spores ◽  
Heterologous Proteins ◽  
Dna Coding ◽  
Gene Coding ◽  
Different Temperatures ◽  
Mucosal Vaccines ◽  
Heterologous Antigens ◽  
Selection Of

Abstract Background: Bacterial spores displaying heterologous antigens or enzymes have long been proposed as mucosal vaccines, functionalized probiotics or biocatalysts. Two main strategies have been developed to display heterologous molecules on the surface of Bacillus subtilis spores: i) a recombinant approach, based on the construction of a gene fusion between a gene coding for a coat protein (carrier) and DNA coding for the protein to be displayed, and ii) a non-recombinant approach, based on the spontaneous and stable adsorption of heterologous molecules on the spore surface. Both systems have advantages and drawbacks and the selection of one or the other depends on the protein to be displayed and on the final use of the activated spore. It has been recently shown that B. subtillis builds structurally and functionally different spores when grown at different temperatures, based on that B. subtilis spores prepared at 25, 37 or 42°C were compared for their efficiency in displaying various model proteins by either the recombinant or the non-recombinant approach.Results: Immune- and fluorescence-based assays were used to analyze the display of several model proteins on spores prepared at 25, 37 or 42°C. Recombinant spores displayed different amounts of the same fusion protein in response to the temperature of spore production. In spores simultaneously displaying two fusion proteins, each of them was differentially displayed at the various temperatures. The display by the non-recombinant approach was only modestly affected by the temperature of spore production, with spores prepared at 37 or 42°C slightly more efficient than 25°C spores in adsorbing at least some of the model proteins tested. Conclusion: Our results indicate that the temperature of spore production allows to control the display of heterologous proteins on spores and, therefore, that the spore-display strategy can be optimized for the specific final use of the activated spores by selecting the display approach, the carrier protein and the temperature of spore production.

scholarly journals Predicting the Structure and Dynamics of Membrane Protein GerAB from Bacillus subtilis

2021 ◽  
Vol 22 (7) ◽  
pp. 3793
Author(s):  
Sophie Blinker ◽  
Jocelyne Vreede ◽  
Peter Setlow ◽  
Stanley Brul
Keyword(s):  
Bacillus Subtilis ◽  
Membrane Protein ◽  
Spore Germination ◽  
Structural Information ◽  
Transmembrane Protein ◽  
Homology Modelling ◽  
Water Channel ◽  
Md Simulations ◽  
Integral Membrane Protein ◽  
Starting Point

Bacillus subtilis forms dormant spores upon nutrient depletion. Germinant receptors (GRs) in spore’s inner membrane respond to ligands such as L-alanine, and trigger spore germination. In B. subtilis spores, GerA is the major GR, and has three subunits, GerAA, GerAB, and GerAC. L-Alanine activation of GerA requires all three subunits, but which binds L-alanine is unknown. To date, how GRs trigger germination is unknown, in particular due to lack of detailed structural information about B subunits. Using homology modelling with molecular dynamics (MD) simulations, we present structural predictions for the integral membrane protein GerAB. These predictions indicate that GerAB is an α-helical transmembrane protein containing a water channel. The MD simulations with free L-alanine show that alanine binds transiently to specific sites on GerAB. These results provide a starting point for unraveling the mechanism of L-alanine mediated signaling by GerAB, which may facilitate early events in spore germination.

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.

Ability of Inactivated Yeast Powder To Adsorb Patulin from Apple Juice

2012 ◽  
Vol 75 (3) ◽  
pp. 585-590 ◽  
Author(s):  
CAIXIA GUO ◽  
TIANLI YUE ◽  
SHAIMAA HATAB ◽  
YAHONG YUAN
Keyword(s):  
Incubation Time ◽  
Apple Juice ◽  
Incubation Temperature ◽  
Absolute Ethyl Alcohol ◽  
Initial Concentration ◽  
Different Temperatures ◽  
Yeast Powder ◽  
Commercial Yeast ◽  
The Stability ◽  
Phosphate Buffered Saline

This study aimed to investigate the adsorption of patulin from apple juice, using two types of inactivated yeast powder: laboratory-prepared yeast powder (LYP) and commercial yeast powder (CYP). The effects of incubation time, pH, incubation temperature, adsorbent amount, and initial concentration of patulin and the stability of the yeast-mycotoxin complex were assessed. The results showed that the efficiencies of the two yeast types in adsorbing patulin were similar. The ability of the powders to remove patulin increased with longer incubation times, and patulin concentration was below detectable levels with LYP and CYP at approximately 36 and 30 h, respectively. The highest removal of patulin was achieved at pH 5.0 for both powder types, and there were no significant differences in patulin decrease at different temperatures (4, 29, and 37°C). Additionally, the adsorption percentage of patulin increased significantly with the increase of absorbent amount and decrease of initial concentration of patulin. Stability of the yeast-patulin complex was assessed, and patulin was more stable when washed in phosphate-buffered saline (pH 4.0) than in absolute ethyl alcohol. These results suggest that inactivated yeast powder has potential as a novel and promising adsorbent to bind patulin effectively.

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