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PubMed1977

Studies on the bacterial spore coat. 6. Effects of alkali extraction on the spore of Bacillus thiaminolyticus.

Thin sections of the spore of Bacillus thiaminolyticus Matsukawa and Misawa show a characteristic surface structure with five ridges, and a series of three distinct layers. The outer layer of the spore coat was peeled off by SDS sonic treatment, and then the middle layer was solubilized by alkali extraction of the SDS sonic-treated spore. The spores subjected to these treatments were still refractile, heat resistant, and contained dipicolinic acid, but lost their resistance to mechanical shock.

Bacillus↗
PubMed2006

The spore photoproduct lyase repairs the 5S- and not the 5R-configured spore photoproduct DNA lesion.

The spore photoproduct lyase is a Fe-S/AdoMet DNA repair enzyme, which directly repairs spore lesions, induced by UV irradiation of spores, using an unknown radical mechanism. The air sensitive radical SAM enzyme was for the first time challenged with synthetically pure substrates. It was found that the enzyme recognizes a synthetic 5S-configured spore lesion without the central phosphodiester bond. The 5R-configured lesion is in contrast to current belief not a substrate.

Bacillus subtilis↗
PubMed1971

"Spore plate method" for transformation of steroids by fungal spores entrapped in silica gel g.

A new technique for investigating steroid biotransformations involving the use of glucose-treated Silica Gel G thin-layer chromatography plates spotted with fungal spores and steroid substrates is described. The conversion is followed by the detection and identification of steroid metabolites and is carried out on single plates by using the spores of different fungi. During the entire process, the spores remain on the original spots and microscopical examination revealed no germination. The method was successfully applied to as little as 30 mug of substrates, and a single plate could be used to detect the steroid metabolizing activity of spores of as many as 15 different cultures.

Journal Article↗
PubMed1987

Lipid composition of Bacillus megaterium spores and spore membranes.

Bacillus megaterium QM B1551 spore lipids were extracted by an improved technique, and the phospholipid and fatty acid compositions were determined. Phospholipids accounted for 65% of the total fatty acids; the neutral lipid fraction contained 15% and the remaining fatty acids were in the interphase, aqueous phase and pellet from the lipid extraction. Each phospholipid had similar fatty acid compositions as did the delipidated pellet. However, the aqueous phase and, to some extent, the interphase had unique fatty acid compositions. Also, fatty acids were found acylated to proteins, which was observed by electrophoresis of delipidated proteins from spores grown in [1-14C]palmitate. Therefore, spores contain unique non-phosphatide fatty acid components that can now be analyzed.

Acylation↗
PubMed1984

Energy-dependent activation of spore-lytic enzyme precursor by germinated spores of Clostridium perfringens.

A precursor of the spore-lytic enzyme of Clostridium perfringens was extracted with alkali from dormant spores of the organism. The enzyme precursor was activated by incubating it with germinated spores which had been treated with alkali. The activation was greatly enhanced by the addition of 3-phosphoglycerate, suggesting that the conversion of precursor to active enzyme depends on endogenous energy-producing metabolism during germination.

Clostridium perfringens↗
PubMed1984

The allergenic and antigenic properties of spore extracts of Aspergillus fumigatus: a comparative study of spore extracts with mycelium and culture filtrate extracts.

The antigenic properties of conidial preparations of Aspergillus fumigatus were compared with mycelial and culture filtrate extracts by use of sera of patients with aspergillosis and hyperimmune rabbit serum. Biologic activities were tested by intracutaneous tests. The immunologic parameters used were: precipitating properties in double-diffusion, IgG binding by ELISA, IgE binding by enzyme inhibition assay, and by enzyme allergosorbent test. It has been demonstrated that the components that are released after disintegration of spores have positive titers in all immunologic assays. The immunologic properties of these spore plasma extracts are comparable with mycelial and/or culture filtrate extracts that are in accordance with the corresponding skin reactivities. Components that are released spontaneously from spores are only weakly positive or negative in the immunologic assays and demonstrate a very low biologic activity.

Allergens↗
PubMed1998

Nucleotide sequence of the sspE genes coding for gamma-type small, acid-soluble spore proteins from the round-spore-forming bacteria Bacillus aminovorans, Sporosarcina halophila and S. ureae.

The single sspE genes coding for gamma-type small, acid-soluble spore proteins (SASP) of three round-spore-forming bacteria, Bacillus aminovorans, Sporosarcina halophila and S. ureae, have been cloned and sequenced. While the deduced amino acid sequences of these three gamma-type SASP show clear homology to those from six Bacillus species that do not form round spores, there are no residues conserved completely among the 9 sequences known. In addition, the 139 residue B. aminovorans protein is 35 residues larger than any other while the 60 residue S. halophila protein is one of the smallest. These data suggest that the sspE genes have been under little selective pressure in recent evolutionary time.

Amino Acid Sequence↗
PubMed2006

Rapid chemical digestion of small acid-soluble spore proteins for analysis of Bacillus spores.

A method for the rapid identification of Bacillus spores is proposed, based on the selective release and chemical digestion of small, acid-soluble spore proteins (SASPs). Microwave-assisted acid hydrolysis of SASPs from B. anthracis str. Sterne and B. subtilis str. 168 was accomplished in a single step requiring only 90 s of heating. The peptide products of the chemical digestion were identified by postsource decay sequencing with a MALDI-TOF-MS equipped with a curved-field reflectron. The specificity of the observed SASP peptides was evaluated using a cross-species sequence search. The incomplete nature of the acid digestion under these conditions allowed detection of the digest products along with the proteins from which they originated, which increased species identification confidence. The feasibility of this approach for the rapid identification of Bacillus species was further demonstrated by analyzing a mixture of B. subtilis str. 168 and B. anthracis str. Sterne spores.

Amino Acid Sequence↗
PubMed1987

Comparative 16S rRNA oligonucleotide analyses and murein types of round-spore-forming bacilli and non-spore-forming relatives.

The phylogenetic incoherency of the genus Bacillus as presently described is demonstrated by analysis of both published and new data from comparative 16S rRNA cataloguing of nine Bacillus species and a number of related non-Bacillus taxa, i.e. Caryophanon latum, Filibacter limicola and Planococcus citreus. While the ellipsoidal-spore-forming bacilli, e.g. B. subtilis and allied species, formed a coherent cluster, the round-spore-forming bacilli showed a higher degree of relationship to the non-spore-forming organisms than these bacilli show among each other. Thus B. sphaericus clustered with C. latum, B. globisporus grouped with F. limicola, B. pasteurii with Sporosarcina ureae, and 'B. aminovorans' with P. citreus, respectively. These organisms formed two related subclusters which, in their phylogenetic depth, are comparable to that of the B. subtilis subline. With the exception of 'B. aminovorans', the 16S rRNA phylogeny was entirely consistent with the distribution of murein types. Even more distantly related to and grouping outside the main Bacillus cluster was B. stearothermophilus, which displayed a moderate relationship to Thermoactinomyces vulgaris. Taxonomic problems arising from the new insights into the intrageneric relationships of Bacillus are discussed.

Bacillus↗
PubMed1996

Immunological detection of the GerA spore germination proteins in the spore integuments of Bacillus subtilis using scanning electron microscopy.

To clarify the molecular mechanisms that trigger spore germination of Bacillus subtilis, the location of GerA proteins (GerAA, GerAB and GerAC), which were reported to be putative gene products of a receptor for one of the germinants, L-alanine, was investigated by immunological techniques using anti-GerA peptide antibodies. Four antibodies were raised against the corresponding epitopes, two in GerAA, one in GerAB and the other in GerAC molecules. The binding of all four antibodies to the inner surface of the cortex-less spore coat fragments could be seen by scanning immunoelectron microscopy with colloidal gold particles. The result agreed with the fact, previously reported, that the colloidal gold particles were visualized just inside the spore coat layer by transmission immunoelectron microscopy using another anti-GerAB peptide antibody.

Antibodies, Bacterial↗
PubMed1988

Decreased UV light resistance of spores of Bacillus subtilis strains deficient in pyrimidine dimer repair and small, acid-soluble spore proteins.

Loss of small, acid-soluble spore protein alpha reduced spore UV resistance 30- to 50-fold in Bacillus subtilis strains deficient in pyrimidine dimer repair, but gave only a 5- to 8-fold reduction in UV resistance in repair-proficient strains. However, both repair-proficient and -deficient spores lacking this protein had identical heat and gamma-radiation resistance.

Bacillus subtilis↗
PubMed2003

Comparison of UV inactivation of spores of three encephalitozoon species with that of spores of two DNA repair-deficient Bacillus subtilis biodosimetry strains.

When exposed to 254-nm UV, spores of Encephalitozoon intestinalis, Encephalitozoon cuniculi, and Encephalitozoon hellem exhibited 3.2-log reductions in viability at UV fluences of 60, 140, and 190 J/m(2), respectively, and demonstrated UV inactivation kinetics similar to those observed for endospores of DNA repair-defective mutant Bacillus subtilis strains used as biodosimetry surrogates. The results indicate that spores of Encephalitozoon spp. are readily inactivated at low UV fluences and that spores of UV-sensitive B. subtilis strains can be useful surrogates in evaluating UV reactor performance.

Animals↗
PubMed1969

Protein synthesis during fungal spore germination. IV. Transfer ribonucleic acid from germinated and ungerminated spores.

Transfer ribonucleic acid (tRNA) fractions isolated from germinated and ungerminated spores of Botryodiplodia theobromae and Rhizopus stolonifer had acceptor activity for all 20 amino acids commonly found in protein, when tested with an enzyme fraction from germinated spores. Accordingly, it is unlikely that the absence of tRNA for a particular amino acid limits protein synthesis in fungal spores.

Amino Acids↗
PubMed1982

Bacillus megaterium spore protease: purification, radioimmunoassay, and analysis of antigen level and localization during growth, sporulation, and spore germination.

The protease which initiates the massive protein degradation early in bacterial spore germination has been purified from Bacillus megaterium spores. The enzyme has a molecular weight of 160,000 and contains four apparently identical subunits, but only the tetramer is enzymatically active. A radioimmunoassay has been developed for this enzyme and has been used to show that the protease is absent from growing cells, but appears early in sporulation within the developing forespore. In contrast, the protease antigen disappears rapidly during spore germination, in parallel with the loss in enzyme activity.

Antigens, Bacterial↗
PubMed1974

Atmospheric pollen and fungal spores in Hamilton in 1972 estimated by the Hirst automatic volumetric spore trap.

A knowledge of the atmospheric pollen and fungal spores is necessary for the diagnosis and management of extrinsic rhinitis and asthma. The Hirst automatic volumetric spore trap has been used for the first time in Canada to identify the quantitative and seasonal incidence of these particles. The trap is easy to operate and has several advantages over the previously used gravity samplers. Tree, grass and ragweed pollens occurred in short, well-defined seasons. Fungal spores greatly outnumbered pollen by 120 to one, and occurred in long, ill-defined seasons. They included large numbers of small basidiospores and ascospores which have previously not been detected in Canada. The latter have not been considered as potential allergens; their clinical importance requires investigation.

Air Movements↗
PubMed1977

[Spore concentration in cities of the European Economic Community. II. Spores of Cladosporium and Alternaria].

A comparative account of Alternaria and Cladosporium spore concentrations in four cities in the European Economic Community (EEC), is given for the years 1973 and 1974. Sijilar volumetric air sampling methods were employed in each of the cities, and concentrations of 3000 Cladosporium spores and 100 Alternaria spores/m3 were taken as the threshold concentrations for clinical significance. With Cladosporium, although highest concentrations were observed in Brussels, the differences were less marked at the other stations. Highest Alternaria concentrations were reocrded in Brussels, followed by London, Leiden and Munich. During the 2 years studied, the average number of days with spore concentrations greater than the threshold value for Cladosporium were 59 in Brussels, 36 in Leiden, 38 in London and 46 in Munich and for Alternaria, 48 in Brussels, 13 in Leiden, 33 in London and 5 in Munich.

Air Pollutants↗
PubMed1984

Bacillus megaterium spore protein C-3: nucleotide sequence of its gene and the amino acid sequence at its spore protease cleavage site.

The nucleotide sequence of the Bacillus megaterium gene coding for spore-specific protein C-3 has been determined. The gene codes for 65 amino acids and the coding sequence is preceded by an efficient ribosome-binding site. The predicted protein C-3 sequence agrees with both the amino acid composition and the amino terminal sequence of protein C-3, and shows homology (approx. 65% of all residues are identical) with the sequences of the analogous proteins A and C of B. megaterium. Protein C-3 is cleaved by the sequence-specific B. megaterium spore protease, and the amino acid sequence at the new amino-terminus generated is identical to that predicted from the gene sequence, and homologous to the spore protease cleavage sites in the A and C proteins. The protein C-3 gene also shares a number of features with the previously sequenced protein C gene in both upstream and downstream flanking sequence.

Amino Acid Sequence↗