K-shell photoionization of ground-state Li-like boron ions [B2+^{2+}]: Experiment and Theory

Absolute cross sections for the K-shell photoionization of ground-state Li-like boron [B$^{2+}$(1s$^2$2s $^2$S)] ions were measured by employing the ion-photon merged-beams technique at the Advanced Light Source synchrotron radiation facility. The energy ranges 197.5--200.5 eV, 201.9--202.1 eV of the [1s(2s\,2p)$^3$P]$^2$P${\rm ^o}$ and [1s(2s\,2p)$^1$P] $^2$P${\rm ^o}$ resonances, respectively, were investigated using resolving powers of up to 17\,600. The energy range of the experiments was extended to about 238.2 eV yielding energies of the most prominent [1s(2$\ell$\,n$\ell^{\prime}$)]$^2$P$^o$ resonances with an absolute accuracy of the order of 130 ppm. The natural linewidths of the [1s(2s\,2p)$^3$P] $^2$P${\rm ^o}$ and [1s(2s\,2p)$^1$P] $^2$P${\rm ^o}$ resonances were measured to be $4.8 \pm 0.6$ meV and $29.7 \pm 2.5$ meV, respectively, which compare favourably with theoretical results of 4.40 meV and 30.53 meV determined using an intermediate coupling R-matrix method.Comment: 6 figures and 2 table

Identification and in vitro Analysis of the GatD/MurT Enzyme-Complex Catalyzing Lipid II Amidation in Staphylococcus aureus

The peptidoglycan of Staphylococcus aureus is characterized by a high degree of crosslinking and almost completely lacks free carboxyl groups, due to amidation of the D-glutamic acid in the stem peptide. Amidation of peptidoglycan has been proposed to play a decisive role in polymerization of cell wall building blocks, correlating with the crosslinking of neighboring peptidoglycan stem peptides. Mutants with a reduced degree of amidation are less viable and show increased susceptibility to methicillin. We identified the enzymes catalyzing the formation of D-glutamine in position 2 of the stem peptide. We provide biochemical evidence that the reaction is catalyzed by a glutamine amidotransferase-like protein and a Mur ligase homologue, encoded by SA1707 and SA1708, respectively. Both proteins, for which we propose the designation GatD and MurT, are required for amidation and appear to form a physically stable bi-enzyme complex. To investigate the reaction in vitro we purified recombinant GatD and MurT His-tag fusion proteins and their potential substrates, i.e. UDP-MurNAc-pentapeptide, as well as the membrane-bound cell wall precursors lipid I, lipid II and lipid II-Gly5. In vitro amidation occurred with all bactoprenol-bound intermediates, suggesting that in vivo lipid II and/or lipid II-Gly5 may be substrates for GatD/MurT. Inactivation of the GatD active site abolished lipid II amidation. Both, murT and gatD are organized in an operon and are essential genes of S. aureus. BLAST analysis revealed the presence of homologous transcriptional units in a number of gram-positive pathogens, e.g. Mycobacterium tuberculosis, Streptococcus pneumonia and Clostridium perfringens, all known to have a D-iso-glutamine containing PG. A less negatively charged PG reduces susceptibility towards defensins and may play a general role in innate immune signaling

Extraction and Analysis of Peptidoglycan Cell Wall Precursors

Extraction and analysis by LC-MS of peptidoglycan precursors represent a valuable method to study antibiotic mode of action and resistance in bacteria. Here, we describe how to apply this method for: (1) testing the action of different classes of antibiotics inhibiting cell wall biosynthesis in Bacillus megaterium; (2) studying the mechanism of self-resistance in mycelial actinomycetes producing glycopeptide antibiotics

Langkocyclines: novel angucycline antibiotics from Streptomyces sp. Acta 3034*

Langkocyclines A1–A3 and B1 and B2, five new angucycline antibiotics produced by Streptomyces sp. Acta 3034, were detected in the course of our HPLC-diode array screening. The producing strain was isolated from the rhizospheric soil of a Clitorea sp. collected from Burau Bay, Langkawi, Malaysia, and was characterized by morphological, physiological and chemotaxonomic features in addition to 16S ribosomal RNA gene sequence information. Strain Acta 3034 is closely related to Streptomyces psammoticus NBRC 13971T and Streptomyces lanatus NBRC 12787T. Langkocyclines consist of an angular tetracyclic benz[a]anthracene skeleton and hydrolyzable O-glycosidic sugar moieties. The yellow-colored A-type langkocyclines differ in their aglycon from the blue-lilac-colored B-type langkocyclines. The A-type langkocycline aglycon is identical to that of aquayamycin and urdamycin A. The chemical structures of the langkocyclines were elucidated by HR-MS, 1D and 2D NMR experiments. They are biologically active against Gram-positive bacteria and exhibit a moderate antiproliferative activity against various human tumor cell lines