Perméation du glycérol et sporulation chez Bacillus subtilis

1972 ◽  
Vol 18 (8) ◽  
pp. 1307-1313 ◽  
Author(s):  
S. A. Saheb
Keyword(s):  
Bacillus Subtilis ◽  
Wild Strain ◽  
Glycerol Kinase ◽  
Simple Diffusion ◽  
Intracellular Pool ◽  
Mutant Cells

Glycerol permeation was studied in B. subtilis 168 trp− gl+ and its two mutants gl−1 and gl−2. The mutant gl−1 has all the enzymes required for the degradation of endocellular glycerol. The mutant gl−2 is impaired in the enzyme glycerol kinase. In the presence of increasing concentrations of glycerol-14C, incorporation of radioactivity by the wild strain obeys Michaelis law while it is linear in the case of the mutant gl−1. Thus the mutant gl−1 is cryptic towards glycerol. No evidence was found as to the formation of an intracellular pool of glycerol in any of the three strains (gl+, gl−1, gl−2) studied. This result suggests that permeation of glycerol in B. subtilis is probably facilitated by diffusion. Sporulation of the mutant gl−1 in presence of glycerol is explained by a mutation affecting the permeation system. Thus glycerol enters in the mutant cells by simple diffusion. As a consequence, the metabolism is greatly reduced and the sporulation mechanism is triggered.

Etude de deux mutants du métabolisme du glycérol chez Bacillus subtilis

1972 ◽  
Vol 18 (8) ◽  
pp. 1315-1325 ◽  
Author(s):  
S. A. Saheb
Keyword(s):  
Bacillus Subtilis ◽  
Carbon Source ◽  
Wild Strain ◽  
Glycerol Kinase ◽  
Glycerol Dehydrogenase ◽  
Dihydroxyacetone Phosphate ◽  
Microbial Species ◽  
Glycerophosphate Dehydrogenase ◽  
Dihydroxyacetone Kinase

As in different microbial species, two pathways for the degradation of glycerol are found in B. subtilis 168. Each pathway includes two enzymes which can catalyze the formation of dihydroxyacetone phosphate from glycerol in vitro. The first pathway includes a glycerol dehydrogenase (gl-D) and a dihydroxyacetone kinase (dha-K). The second pathway includes a glycerol kinase (gl-K) and an α-glycerophosphate dehydrogenase (glp-D). Enzymes of both pathways are repressed in the presence of glucose. Only the enzymes of the second pathway are inducible. The inducer is probably glycerophosphate, utilization of which as a carbon source by B. subtilis is demonstrated. Degradation of glycerol in B. subtilis proceeds through the second pathway. This was demonstrated by the isolation of a mutant (gl−2) impaired in glycerol kinase, and which cannot use glycerol as a carbon source. Another mutant (gl−1)was isolated, which cannot use glycerol as a carbon source. When comparing the activity of the four enzymes, particularly gl-K, no significant differences were observed between the wild strain and the mutant gl−1. This leads one to consider the existence of a glycerol permeation system in B. subtilis. A mutation affecting this system would explain the behavior of the mutant gl−1.

Antagonistic activity of Bacillus subtilis strains isolated from various sources

2018 ◽  
Vol 8 (2) ◽  
pp. 354-364
Author(s):  
A. N. Irkitova ◽  
A. V. Grebenshchikova ◽  
A. V. Matsyura
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Wild Strain ◽  
Fish Farming ◽  
Antagonistic Activity ◽  
Antagonistic Effect ◽  
E Coli ◽  
Long Period ◽  
Test Culture ◽  
Agar Media

<p>An important link in solving the problem of healthy food is the intensification of the livestock, poultry and fish farming, which is possible only in the adoption and rigorous implementation of the concept of rational feeding of animals. In the implementation of this concept required is the application of probiotic preparations. Currently, there is an increased interest in spore probiotics. In many ways, this can be explained by the fact that they use no vegetative forms of the bacilli and their spores. This property provides spore probiotics a number of advantages: they are not whimsical, easily could be selected, cultivated, and dried. Moreover, they are resistant to various factors and could remain viable during a long period. One of the most famous spore microorganisms, which are widely used in agriculture, is <em>Bacillus subtilis</em>. Among the requirements imposed to probiotic microorganisms is mandatory – antagonistic activity to pathogenic and conditional-pathogenic microflora. The article presents the results of the analysis of antagonistic activity of collection strains of <em>B. subtilis</em>, and strains isolated from commercial preparations. We studied the antagonistic activity on agar and liquid nutrient medias to trigger different antagonism mechanisms of <em>B. subtilis</em>. On agar media, we applied three diffusion methods: perpendicular bands, agar blocks, agar wells. We also applied the method of co-incubating the test culture (<em>Escherichia coli</em>) and the antagonist (or its supernatant) in the nutrient broth. Our results demonstrated that all our explored strains of <em>B. subtilis</em> have antimicrobial activity against a wild strain of <em>E. coli</em>, but to varying degrees. We identified strains of <em>B. subtilis</em> with the highest antagonistic effect that can be recommended for inclusion in microbial preparations for agriculture.</p><p><em><br /></em><em></em></p>

scholarly journals Localization of Cold Shock Proteins to Cytosolic Spaces Surrounding Nucleoids in Bacillus subtilis Depends on Active Transcription

2001 ◽  
Vol 183 (21) ◽  
pp. 6435-6443 ◽  
Author(s):  
Michael H. W. Weber ◽  
Arsen V. Volkov ◽  
Ingo Fricke ◽  
Mohamed A. Marahiel ◽  
Peter L. Graumann
Keyword(s):  
Bacillus Subtilis ◽  
Cold Shock ◽  
Fluorescent Protein ◽  
Cold Shock Protein ◽  
Initiation Of Translation ◽  
Specific Localization ◽  
Active Transcription ◽  
Green Fluorescent ◽  
Shock Proteins ◽  
Mutant Cells

ABSTRACT Using immunofluorescence microscopy and a fusion of a cold shock protein (CSP), CspB, to green fluorescent protein (GFP), we showed that in growing cells Bacillus subtilis CSPs specifically localize to cytosolic regions surrounding the nucleoid. The subcellular localization of CSPs is influenced by the structure of the nucleoid. Decondensed chromosomes in smc mutant cells reduced the sizes of the regions in which CSPs localized, while cold shock-induced chromosome compaction was accompanied by an expansion of the space in which CSPs were present. As a control, histone-like protein HBsu localized to the nucleoids, while β-galactosidase and GFP were detectable throughout the cell. After inhibition of translation, CspB-GFP was still present around the nucleoids in a manner similar to that in cold-shocked cells. However, in stationary-phase cells and after inhibition of transcription, CspB was distributed throughout the cell, indicating that specific localization of CspB depends on active transcription and is not due to simple exclusion from the nucleoid. Furthermore, we observed that nucleoids are more condensed and frequently abnormal incspB cspC and cspB cspDdouble-mutant cells. This suggests that the function of CSPs affects chromosome structure, probably through coupling of transcription to translation, which is thought to decondense nucleoids. In addition, we found that cspB cspD and cspB cspC double mutants are defective in sporulation, with a block at or before stage 0. Interestingly, CspB and CspC are depleted from the forespore compartment but not from the mother cell. In toto, our findings suggest that CSPs localize to zones of newly synthesized RNA, coupling transcription with initiation of translation.

scholarly journals Regulation of a New Cell Wall Hydrolase Gene,cwlF, Which Affects Cell Separation in Bacillus subtilis

1998 ◽  
Vol 180 (9) ◽  
pp. 2549-2555 ◽  
Author(s):  
Shu Ishikawa ◽  
Yoshiko Hara ◽  
Ryo Ohnishi ◽  
Junichi Sekiguchi
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Rna Polymerase ◽  
Sequence Similarity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Exponential Growth Phase ◽  
High Sequence Similarity ◽  
Cell Wall Hydrolase ◽  
Mutant Cells

ABSTRACT Bacillus subtilis produces a 35-kDa cell wall hydrolase, CwlF, during vegetative growth. The CwlF protein was extracted from B. subtilis cwlB sigD mutant cells and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. N-terminal amino acid sequencing revealed that its sequence is completely identical to that of the internal region of thepapQ gene product. Disruption of the papQ gene in the B. subtilis chromosome led to the complete loss of CwlF, indicating that papQ is identical tocwlF. CwlF exhibits high sequence similarity to the p60 proteins of Listeria species, NlpC proteins ofEscherichia coli and Haemophilus influenzae, and Enp2 protein of Bacillus sphaericus. The β-galactosidase activity of the cwlF-lacZ transcriptional fusion and Northern blot analysis of the cwlF gene indicated that the gene is expressed as a monocistronic operon during the exponential growth phase, and primer extension analysis suggested that the cwlF gene is transcribed mainly by EςA RNA polymerase and weakly by EςH RNA polymerase. While the cells of the cwlF-deficient mutant were about twice as long as those of the wild-type strain, the cwlF sigD double mutant cells exhibited extraordinary microfiber formation, in contrast to the filamentation of the sigD mutant. The CwlF production was not affected by the pleiotropic mutationsflaD1 and degU32(Hy), which endow cells with the ability of extensive filamentation.

scholarly journals Cardiolipin Domains in Bacillus subtilis Marburg Membranes

2004 ◽  
Vol 186 (5) ◽  
pp. 1475-1483 ◽  
Author(s):  
Fumitaka Kawai ◽  
Momoko Shoda ◽  
Rie Harashima ◽  
Yoshito Sadaie ◽  
Hiroshi Hara ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Lipid Composition ◽  
Exponential Growth Phase ◽  
Dye Molecules ◽  
Wild Type ◽  
Green Fluorescence ◽  
Paralogous Genes ◽  
Sporulation Initiation ◽  
Mutant Cells ◽  
Sporulation Phase

ABSTRACT Recently, use of the cardiolipin (CL)-specific fluorescent dye 10-N-nonyl-acridine orange (NAO) revealed CL-rich domains in the Escherichia coli membrane (E. Mileykovskaya and W. Dowhan, J. Bacteriol. 182: 1172-1175, 2000). Staining of Bacillus subtilis cells with NAO showed that there were green fluorescence domains in the septal regions and at the poles. These fluorescence domains were scarcely detectable in exponentially growing cells of the clsA-disrupted mutant lacking detectable CL. In sporulating cells with a wild-type lipid composition, fluorescence domains were observed in the polar septa and on the engulfment and forespore membranes. Both in the clsA-disrupted mutant and in a mutant with disruptions in all three of the paralogous genes (clsA, ywjE, and ywiE) for CL synthase, these domains did not vanish but appeared later, after sporulation initiation. A red shift in the fluorescence due to stacking of two dye molecules and the lipid composition suggested that a small amount of CL was present in sporulating cells of the mutants. Mass spectrometry analyses revealed the presence of CL in these mutant cells. At a later stage during sporulation of the mutants the frequency of heat-resistant cells that could form colonies after heat treatment was lower. The frequency of sporulation of these cells at 24 h after sporulation initiation was 30 to 50% of the frequency of the wild type. These results indicate that CL-rich domains are present in the polar septal membrane and in the engulfment and forespore membranes during the sporulation phase even in a B. subtilis mutant with disruptions in all three paralogous genes, as well as in the membranes of the medial septa and at the poles during the exponential growth phase of wild-type cells. The results further suggest that the CL-rich domains in the polar septal membrane and engulfment and forespore membranes are involved in sporulation.

scholarly journals FlhF, the Third Signal Recognition Particle-GTPase of Bacillus subtilis, Is Dispensable for Protein Secretion

2004 ◽  
Vol 186 (17) ◽  
pp. 5956-5960 ◽  
Author(s):  
Geeske Zanen ◽  
Haike Antelmann ◽  
Helga Westers ◽  
Michael Hecker ◽  
Jan Maarten van Dijl ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Protein Secretion ◽  
Signal Recognition Particle ◽  
Signal Recognition ◽  
The Third ◽  
Mutant Cells

ABSTRACT Bacillus subtilis contains three proteins of the signal recognition particle-GTPase family known as Ffh, FtsY, and FlhF. Here we show that FlhF is dispensable for protein secretion, unlike Ffh and FtsY. Although flhF is located in the fla/che operon, B. subtilis 168 flhF mutant cells assemble flagella and are motile.

scholarly journals The Composition of the Cell Envelope Affects Conjugation in Bacillus subtilis

2016 ◽  
Vol 198 (8) ◽  
pp. 1241-1249 ◽  
Author(s):  
Christopher M. Johnson ◽  
Alan D. Grossman
Keyword(s):  
Bacillus Subtilis ◽  
Gene Transfer ◽  
Horizontal Gene Transfer ◽  
Cell Envelope ◽  
Microbial Evolution ◽  
Major Type ◽  
Content Type ◽  
New Genes ◽  
Mutant Cells ◽  
Integrative And Conjugative Element

ABSTRACTConjugation in bacteria is the contact-dependent transfer of DNA from one cell to another via donor-encoded conjugation machinery. It is a major type of horizontal gene transfer between bacteria. Conjugation of the integrative and conjugative element ICEBs1intoBacillus subtilisis affected by the composition of phospholipids in the cell membranes of the donor and recipient. We found that reduction (or elimination) of lysyl-phosphatidylglycerol caused by loss ofmprFcaused a decrease in conjugation efficiency. Conversely, alterations that caused an increase in lysyl-phosphatidylglycerol, including loss ofugtPor overproduction ofmprF, caused an increase in conjugation efficiency. In addition, we found that mutations that alter production of other phospholipids, e.g., loss ofclsAandyfnI, also affected conjugation, apparently without substantively altering levels of lysyl-phosphatidylglycerol, indicating that there are multiple pathways by which changes to the cell envelope affect conjugation. We found that the contribution ofmprFto conjugation was affected by the chemical environment. Wild-type cells were generally more responsive to addition of anions that enhanced conjugation, whereasmprFmutant cells were more sensitive to combinations of anions that inhibited conjugation at pH 7. Our results indicate thatmprFand lysyl-phosphatidylglycerol allow cells to maintain relatively consistent conjugation efficiencies under a variety of ionic conditions.IMPORTANCEHorizontal gene transfer is a driving force in microbial evolution, enabling cells that receive DNA to acquire new genes and phenotypes. Conjugation, the contact-dependent transfer of DNA from a donor to a recipient by a donor-encoded secretion machine, is a prevalent type of horizontal gene transfer. Although critically important, it is not well understood how the recipient influences the success of conjugation. We found that the composition of phospholipids in the membranes of donors and recipients influences the success of transfer of the integrative and conjugative element ICEBs1inBacillus subtilis. Specifically, the presence of lysyl-phosphatidylglycerol enables relatively constant conjugation efficiencies in a range of diverse chemical environments.

scholarly journals Phosphatidylethanolamine Domains and Localization of Phospholipid Synthases in Bacillus subtilis Membranes

2005 ◽  
Vol 187 (6) ◽  
pp. 2163-2174 ◽  
Author(s):  
Ayako Nishibori ◽  
Jin Kusaka ◽  
Hiroshi Hara ◽  
Masato Umeda ◽  
Kouji Matsumoto
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Green Fluorescent Protein ◽  
Acridine Orange ◽  
Fluorescent Protein ◽  
Cyclic Peptide ◽  
Peptide Probe ◽  
Phosphatidylserine Synthase ◽  
Green Fluorescent ◽  
Mutant Cells

ABSTRACT Application of the cardiolipin (CL)-specific fluorescent dye 10-N-nonyl-acridine orange has recently revealed CL-rich domains in the septal regions and at the poles of the Bacillus subtilis membrane (F. Kawai, M. Shoda, R. Harashima, Y. Sadaie, H. Hara, and K. Matsumoto, J. Bacteriol. 186:1475-1483, 2004). This finding prompted us to examine the localization of another phospholipid, phosphatidylethanolamine (PE), with the cyclic peptide probe, Ro09-0198 (Ro), that binds specifically to PE. Treatment with biotinylated Ro followed by tetramethyl rhodamine-conjugated streptavidin revealed that PE is localized in the septal membranes of vegetative cells and in the membranes of the polar septum and the engulfment membranes of sporulating cells. When the mutant cells of the strains SDB01 (psd1::neo) and SDB02 (pssA10::spc), which both lack PE, were examined under the same conditions, no fluorescence was observed. The localization of the fluorescence thus evidently reflected the localization of PE-rich domains in the septal membranes. Similar PE-rich domains were observed in the septal regions of the cells of many Bacillus species. In Escherichia coli cells, however, no PE-rich domains were found. Green fluorescent protein fusions to the enzymes that catalyze the committed steps in PE synthesis, phosphatidylserine synthase, and in CL synthesis, CL synthase and phosphatidylglycerophosphate synthase, were localized mainly in the septal membranes in B. subtilis cells. The majority of the lipid synthases were also localized in the septal membranes; this includes 1-acyl-glycerol-3-phosphate acyltransferase, CDP-diacylglycerol synthase, phosphatidylserine decarboxylase, diacylglycerol kinase, glucolipid synthase, and lysylphosphatidylglycerol synthase. These results suggest that phospholipids are produced mostly in the septal membranes and that CL and PE are kept from diffusing out to lateral ones.

scholarly journals Bex, the Bacillus subtilis Homolog of the Essential Escherichia coli GTPase Era, Is Required for Normal Cell Division and Spore Formation

2002 ◽  
Vol 184 (22) ◽  
pp. 6389-6394 ◽  
Author(s):  
Natalie Minkovsky ◽  
Arash Zarimani ◽  
Vasant K. Chary ◽  
Brian H. Johnstone ◽  
Bradford S. Powell ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Cell Division ◽  
Normal Cell ◽  
Spore Formation ◽  
E Coli ◽  
Mutant Cells

ABSTRACT The Bacillus subtilis bex gene complemented the defect in an Escherichia coli era mutant. The Bex protein showed 39% identity and 67% similarity to the E. coli Era GTPase. In contrast to era, bex was not essential in all strains. bex mutant cells were elongated and filled with diffuse nucleoid material. They grew slowly and exhibited severely impaired spore formation.

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