J Microbiol. 2011 Oct ;49 (5):776-81 22068494
Department of Bioengineering, Granduate School of Kyonggi University, Suwon, 443-760, Republic of Korea.
A Gram-positive, rod-shaped, motile, endospore-forming bacterial strain, designated NB22(T), was isolated from soil of a lettuce field in Kyonggi province, South Korea, and was characterized by using a polyphasic taxonomic approach. This novel isolate grew optimally at 30-37°C and pH 8-9. It grew in the presence of 0-4% NaCl (optimum, 1-2%). Comparative 16S rRNA gene sequence analysis showed that strain NB22(T) was closely related to members of the genus Bacillus and fell within a coherent cluster comprising B. siralis 171544(T)(98.1%) and B. korlensis ZLC-26(T)(97.3%). The levels of 16S rRNA gene sequence similarity with respect to other Bacillus species with validly published names were less than 96.4%. Strain NB22(T) had a genomic DNA G+C content of 36.3 mol% and the predominant respiratory quinone was MK-7. The peptidoglycan contained meso-diaminopimelic acid. The major cellular fatty acids were iso-C(15:0), anteiso-C(15:0), C(14:0), and C(16:0). These chemotaxonomic results supported the affiliation of strain NB22(T) to the genus Bacillus, and the low DNA-DNA relatedness values and distinguishing phenotypic characteristics allowed genotypic and phenotypic differentiation of strain NB22(T) from recognized Bacillus species. On the basis of the evidence presented, strain NB22(T) is considered to represent a novel species of the genus Bacillus, for which the name Bacillus kyonggiensis sp. nov. is proposed. The type strain is NB22(T)(=KEMB 5401-267(T)=JCM 17569(T)).
Most cited papers:
Tamaki Yano, Shizuka Mita, Hiroko Ohmori, Yoshiteru Oshima, Yukari Fujimoto, Ryu Ueda, Haruhiko Takada, William E Goldman, Koichi Fukase, Neal Silverman, Tamotsu Yoshimori, Shoichiro Kurata
Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai 980-8578, Japan.
Autophagy, an evolutionally conserved homeostatic process for catabolizing cytoplasmic components, has been linked to the elimination of intracellular pathogens during mammalian innate immune responses. However, the mechanisms underlying cytoplasmic infection-induced autophagy and the function of autophagy in host survival after infection with intracellular pathogens remain unknown. Here we report that in drosophila, recognition of diaminopimelic acid-type peptidoglycan by the pattern-recognition receptor PGRP-LE was crucial for the induction of autophagy and that autophagy prevented the intracellular growth of Listeria monocytogenes and promoted host survival after this infection. Autophagy induction occurred independently of the Toll and IMD innate signaling pathways. Our findings define a pathway leading from the intracellular pattern-recognition receptors to the induction of autophagy to host defense.
Center for Advanced Research in Biotechnology, W. M. Keck Laboratory for Structural Biology, University of Maryland Biotechnology Institute, Rockville, MD 20850, USA.
Peptidoglycan (PGN) recognition proteins (PGRPs) are pattern-recognition receptors of the innate immune system that bind and, in some cases, hydrolyze bacterial PGNs. We determined the crystal structure, at 2.30-A resolution, of the C-terminal PGN-binding domain of human PGRP-Ialpha in complex with a muramyl tripeptide representing the core of lysine-type PGNs from Gram-positive bacteria. The peptide stem of the ligand is buried at the deep end of a long binding groove, with N-acetylmuramic acid situated in the middle of the groove, whose shallow end can accommodate a linked N-acetylglucosamine. Although most interactions are with the peptide, the glycan moiety also seems to be essential for specific recognition by PGRPs. Conservation of key PGN-contacting residues shows that all PGRPs employ this basic PGN-binding mode. The structure pinpoints variable residues that likely mediate discrimination between lysine- and diaminopimelic acid-type PGNs. We also propose a mechanism for PGN hydrolysis by Zn(2+)-containing PGRPs.
Salivary gland homogenates from adult Aedes aegypti lyse Micrococcus lysodeikticus cells. The bacteriolytic factor is present in a cell type common to both male and female mosquitoes, as well as in the crop of sugar-feeding mosquitoes. The bacteriolytic factor releases digestion products from sacculi of Escherichia coli that are different from those of hen egg white lysozyme.
Stable L-phase variants isolated from Bacillus licheniformis and Bacillus subtilis, when grown in osmotically stabilized media, do not synthesize peptidoglycan but have been found to accumulate the nucleotide precursors of this polymer. The enzymes involved in the synthesis of these precursors and the later membrane-bound stages of peptidoglycan synthesis have been investigated, and the L-phase variants have been shown to contain lesions, which provide a rational explanation for the absence of peptidoglycan and for the nature of the precursor accumulated. The majority of the L-phase variants contained a single enzymic defect, but two strains were isolated with double lesions. Five out of seven strains examined accumulated uridine 5'-diphosphate (UDP)-MurAc-L-ala-D-glu and were unable to synthesize diaminopimelic acid as a consequence of a defect in aspartate-beta-semialdehyde dehydrogenase activity. Two strains were deficient in UDP-MurAc: L-alanine ligase and accumulated UDP-MurAc. One strain accumulated the complete nucleotide precursor UDP-MurAc-L-ala-D-glu-mA2pm-D-ala-D-ala and was deficient in phospho-N-acetylmuramyl pentapeptide translocase. A second strain also had this lesion, together with defective aspartate-beta-semialdehyde dehydrogenase activity. The other enzymes of peptidoglycan synthesis were present in the L-phase variants, with activities similar to those found in the parent bacilli grown under identical conditions. Membrane preparations from certain of the L-phase variants were also capable of synthesizing the secondary polymers poly(glycerol phosphate) teichoic acid and teichuronic acid and also a polymer of N-acetylglucosamine.
A high activity of meso-alpha-epsilon-diaminopimelate dehydrogenase was found in extracts of Bacillus sphaericus, Brevibacterium sp., Corynebacterium glutamicum, and Proteus vulgaris among bacteria tested. B. sphaericus IFO 3525, in which the enzyme is most abundant, was chosen to study the enzyme reaction. The enzyme was not induced by the addition of meso-alpha-epsilon-diaminopimelate to the growth medium. The reaction product was isolated and identified as alpha-amino-epsilon-ketopimelate by a comparison of the properties of its 2,4-dinitrophenylhydrazone with those of an authentic sample in silica gel thin-layer chromatography, absorption, infrared and proton nuclear magnetic resonance spectrometry, and elemental analyses. The alpha-amino-epsilon-ketopimelate formed enzymatically was decarboxylated by H2O2 to yield L-alpha-aminoadipate. This suggests that the amino group with D-configuration in the substrate is oxidatively deaminated; the enzyme is a D-amino acid dehydrogenase. L-alpha-Amino-epsilon-ketopimelate undergoes spontaneous dehydration to the cyclic delta1-piperideine-2,6-dicarboxylate. The enzyme reaction is reversible, and meso-alpha-epsilon-diaminopimelate was formed in the reductive amination of L-alpha-epsilon-ketopimelate.
Department of Microbiology, University of Guelph, Ontario, Canada.
A high-performance anion-exchange chromatography method with pulsed-amperometric detection has been developed for the simultaneous analysis of both amino acids and amino sugars and applied to the compositional analysis of peptidoglycan hydrolysates. Chromatography of the acid hydrolysis products was performed on a CarboPac PA-1 anion-exchange column, with pulsed-amperometric detection. Complete resolution of the two amino sugars (glucosamine and muramic acid) and eight of the nine amino acids (Ala, diaminobutyric acid, diaminopimelic acid, Glu, Gly, homoserine, Lys, Orn, and Ser) known to occur in various peptidoglycans was achieved within 70 min. Only homoserine and glycine (retention times 26.8 and 26.9 min, respectively) were not resolved by this procedure, but the simultaneous occurrence of these two amino acids in peptidoglycan is extremely rare. Reproducibility of the separations was shown to be very high and detection limits exceeded 10 pmol for glucosamine. This convenient and simple analysis was applied to the quantitation of many crude peptidoglycan samples isolated from the species of the Proteeae (Proteus, Providencia, and Morganella) for the determination of the extent of peptidoglycan O-acetylation.
Purification of penicillin-insensitive DD-endopeptidase, a new cell wall peptidoglycan-hydrolyzing enzyme in Escherichia coli, and its inhibition by deoxyribonucleic acids.
Institute for Fermentation, Osaka, Japan. email@example.com
Phylogenetic analysis of 'Actinoalloteichus cyanogriseus' based on its 16S rDNA sequence confirmed that the organism belongs to the family Pseudonocardiaceae. It contains glutamic acid, alanine and meso-diaminopimelic acid as cell wall amino acids, and menaquinone 9 (H4). Its taxonomic characteristics differ from those of the previously described organisms of the family Pseudonocardiaceae. On the basis of these morphological, physiological, chemotaxonomic and phylogenetic analyses, a new genus, Actinoalloteichus, is proposed, along with a new species, namely Actinoalloteichus cyanogriseus sp. nov. The type strain is A. cyanogriseus IFO 14455T (= AS 4.1159T = JCM 6095T).
Stability of a host-vector system based on complementation of an essential gene in Escherichia coli.
Department of Molecular Biology, Transgene S.A., Strasbourg, France.
Antibiotic selection is the most common selection system for plasmid-containing bacteria. This technique, nevertheless, can be a source of problems during the expression of heterologous genes in Escherichia coli. We have developed an alternative selection system based on the complementation of a chromosomal auxotrophic (dapD2) mutation by the corresponding wild type gene carried on a plasmid. We show that the system effectively selects for the presence of plasmid on solid and liquid medium. In addition, we have observed a loss of viability associated with high levels of gene expression and accumulation of a heterologous protein, but the selective power and improved intrinsic stability of the dap+ plasmid, compared to a beta-lactamase (bla) based vector, excludes overgrowth of the culture by plasmidless cells.
Substrate specificity of Escherichia coli LD-carboxypeptidase on biosynthetically modified muropeptides.
Centro de Biologia Molecular Severo Ochoa CSIC-UAM, Facultad de Ciencias UAM, Madrid, Spain.
Escherichia coli murein can be biosynthetically modified. Amino acids at positions 3 and 4 (m-diaminopimelic acid and D-alanine, respectively) on the peptide moieties can be changed under appropriate growth conditions. The activity of E. coli LD-carboxypeptidase on biosynthetically modified substrates has been studied in vitro. The enzyme hydrolysed all tested disaccharide-tetrapeptide monomeric muropeptides modified at position 4. Monomers with m-lanthionine, but not with L-ornithine, instead of m-diaminopimelic acid at position 3 were accepted. However, neither cross-linked muropeptides nor macromolecular murein were substrates for the reaction. Our observations argue against a direct effect of LD-carboxypeptidase on macromolecular murein metabolism.