Delayed Germination of Bacillus cereus T Spores after Treatment with Trichloroacetic Acid and Their Reactivation by Heating

Shibata, Hirofumi; Murakami, Hiromi; Tani, Isamu

doi:10.1111/j.1348-0421.1980.tb02832.x

Cited by 3 publications

(1 citation statement)

References 11 publications

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“…Another explanation is that single L-alanine residues, the low number of cross-links compared to those in vegetative peptidoglycan, and the absence of amidation on the D-carboxyl group of diaminopimelic acid account for the fact that the cortical peptidoglycan is more negatively charged than vegetative peptidoglycan (6). It has been suggested that this high net negative charge may be essential for the function of the cortex in heat resistance (19). The lytH cwlD mutant exhibits the simplest peptidoglycan profile of any strain so far examined by muropeptide analysis.…”

Section: Discussionmentioning

confidence: 99%

Characterization of LytH, a Differentiation-Associated Peptidoglycan Hydrolase of Bacillus subtilis Involved in Endospore Cortex Maturation

2003

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The cortex peptidoglycan from endospores of Bacillus subtilis is responsible for the maintenance of dormancy. LytH (YunA) has been identified as a novel sporulation-specific component with a role in cortex structure determination. The lytH gene was expressed only during sporulation, under the control of the mother cellspecific sigma factor K . Spores of a lytH mutant have slightly reduced heat resistance and altered staining when viewed by electron microscopy. Analysis of the peptidoglycan structure of lytH mutant spores shows the loss of muramic acid residues substituted with L-alanine and a corresponding increase in muramic acid residues substituted with tetrapeptide compared to those in the parent strain. In a lytH cwlD mutant, the lack of muramic acid residues substituted with L-alanine and ␦-lactam leaves 97% of residues substituted with tetrapeptide. These results suggest that lytH encodes an L-Ala-D-Glu peptidase involved in production of single L-alanine side chains from tetrapeptides in the spore cortex. The lack of di-or tripeptides in a lytH mutant reveals the enzyme is an endopeptidase.Bacterial endospores are characterized by their extreme dormancy and high-level resistance to a range of stresses, in particular heat (7). The specialized spore structure determines resistance properties, with the dehydrated spore core being crucial for heat resistance. The spore cell wall peptidoglycan, known as the spore cortex, is essential for the maintenance of the dehydrated core, heat resistance, and dormancy (7). The cortex has a unique, spore-specific structure, and recent analysis has begun to identify features important in its role during differentiation and to reveal components responsible for cortex synthesis, modification during sporulation, and hydrolysis during germination (1, 2). Spore cortex is characterized by an extremely low level of cross-linking, which is important for the maintenance of heat resistance (1, 15). A unique feature of the dormant spore cortex is the presence of muramic acid ␦-lactam residues. This modification has a specific role during germination, because the putative germination-specific lytic enzymes SleB and CwlJ, which are responsible for cortex degradation, require the presence of muramic acid ␦-lactam for substrate recognition, since cortex lacking this modification is unable to be hydrolyzed during germination (2).During cortex maturation, a sporulation-specific amidase, CwlD, and a recently described polysaccharide deacetylase, PdaA, are responsible for the formation of the characteristic ␦-lactam residues of bacterial endospores (1,8,15). Although cwlD or pdaA mutants and a cwlD pdaA double mutant can produce resistant endospores, no cortex hydrolysis is observed during germination, and the spores do not outgrow (1,2,8,16,18). The final characteristic feature of the cortex is that about 25% of the muramic acid side chains are substituted for with single L-alanine residues (1, 17). These may be formed by the action of an L-Ala-D-Glu endopeptidase on nascent stem peptides. ...

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Section: Discussionmentioning

confidence: 99%

Characterization of LytH, a Differentiation-Associated Peptidoglycan Hydrolase of Bacillus subtilis Involved in Endospore Cortex Maturation

2003

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Pilot-scale investigation of sludge reduction in aerobic digestion system with endospore-forming bacteria

Seo

Choi

et al. 2017

Chemosphere

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Development of a Method for the Extraction of the Total Proteome of Bacillus anthracis Spores

Koteneva

Tsygankova

Калинин

et al. 2021

BMCRM

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We have developed a method for obtaining the total proteome of Bacillus anthracis spores, which combines efficient protein extraction with reliable disinfection of samples. We used 7 strains of B. anthracis: 4 plasmid-free, 3 diplasmid, one of which with an atypical type of capsule formation in air. The schemes for isolating the total spore proteome were tested using various lysis solutions in the presence of bacterial protease inhibitors, with the inclusion of the stages of treatment of spores with trichloroacetic acid and mechanical disintegration and with their exclusion. The quality and completeness of the extraction of the total proteome of the spores of the samples was assessed in one-dimensional (1D) and twodimensional (2D) electrophoresis. Treatment of spores with trichloroacetic acid increased the reliability of material disinfection and reduces the loss of the final product. Mechanical disintegration after treatment of spores with lysing solutions increases the completeness of extraction of spore proteins in a wide range of molecular weights and facilitated the process of sterilizing filtration. Final filtration of the lysate through a PVDF filter with a pore size of 0.22 μm provided additional decontamination of samples without reducing their quality. Thus, the use of the proposed sample preparation scheme makes it possible to obtain complete protein extracts of spores of B. anthracis strains, suitable for comparative analysis of the proteome and search for a possible correlation with the features phenotypic properties and mechanisms that ensure the preservation of the pathogen anthrax in environmental objects, including soil

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Delayed Germination of Bacillus cereus T Spores after Treatment with Trichloroacetic Acid and Their Reactivation by Heating

Cited by 3 publications

References 11 publications

Characterization of LytH, a Differentiation-Associated Peptidoglycan Hydrolase of Bacillus subtilis Involved in Endospore Cortex Maturation

Characterization of LytH, a Differentiation-Associated Peptidoglycan Hydrolase of Bacillus subtilis Involved in Endospore Cortex Maturation

Pilot-scale investigation of sludge reduction in aerobic digestion system with endospore-forming bacteria

Development of a Method for the Extraction of the Total Proteome of Bacillus anthracis Spores

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