Purification and biochemical characterisation of GlmU from Yersinia pestis

Patin, Delphine; Bayliss, Marc; Mengin‐Lecreulx, Dominique; Oyston, Petra C. F.; Blanot, Didier

doi:10.1007/s00203-014-1065-0

Cited by 6 publications

(3 citation statements)

References 44 publications

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“…The bifunctional N -acetyltransferase/uridylyltransferase GlmU (CDIF630erm_03829) catalyzes the transfer of an acetyl from acetyl-coenzyme A to glucosamine 1-phosphate to form N -acetylglucosamine 1-phosphate during cell wall biosynthesis. The protein is necessary for the infection of various pathogenic bacteria, including Mycobacterium tuberculosis , Yersinia pestis , Haemophilus influenzae and Xanthomonas oryzae (Buurman et al, 2011; Min et al, 2012; Patin et al, 2015), it serves as target for the antimicrobial treatment of Mycobacteria (Sharma and Khan, 2017). The glmU gene obviously forms an operon with the prs gene (CDIF630erm_03828) encoding ribose-phosphate pyrophosphokinase that catalyzes the conversion of ribose-5-phosphate into phosphoribosyl pyrophosphate during nucleotide biosynthesis.…”

Section: Resultsmentioning

confidence: 99%

Iron Regulation in Clostridioides difficile

et al. 2018

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The response to iron limitation of several bacteria is regulated by the ferric uptake regulator (Fur). The Fur-regulated transcriptional, translational and metabolic networks of the Gram-positive, pathogen Clostridioides difficile were investigated by a combined RNA sequencing, proteomic, metabolomic and electron microscopy approach. At high iron conditions (15 μM) the C. difficile fur mutant displayed a growth deficiency compared to wild type C. difficile cells. Several iron and siderophore transporter genes were induced by Fur during low iron (0.2 μM) conditions. The major adaptation to low iron conditions was observed for the central energy metabolism. Most ferredoxin-dependent amino acid fermentations were significantly down regulated (had, etf, acd, grd, trx, bdc, hbd). The substrates of these pathways phenylalanine, leucine, glycine and some intermediates (phenylpyruvate, 2-oxo-isocaproate, 3-hydroxy-butyryl-CoA, crotonyl-CoA) accumulated, while end products like isocaproate and butyrate were found reduced. Flavodoxin (fldX) formation and riboflavin biosynthesis (rib) were enhanced, most likely to replace the missing ferredoxins. Proline reductase (prd), the corresponding ion pumping RNF complex (rnf) and the reaction product 5-aminovalerate were significantly enhanced. An ATP forming ATPase (atpCDGAHFEB) of the F0F1-type was induced while the formation of a ATP-consuming, proton-pumping V-type ATPase (atpDBAFCEKI) was decreased. The [Fe-S] enzyme-dependent pyruvate formate lyase (pfl), formate dehydrogenase (fdh) and hydrogenase (hyd) branch of glucose utilization and glycogen biosynthesis (glg) were significantly reduced, leading to an accumulation of glucose and pyruvate. The formation of [Fe-S] enzyme carbon monoxide dehydrogenase (coo) was inhibited. The fur mutant showed an increased sensitivity to vancomycin and polymyxin B. An intensive remodeling of the cell wall was observed, Polyamine biosynthesis (spe) was induced leading to an accumulation of spermine, spermidine, and putrescine. The fur mutant lost most of its flagella and motility. Finally, the CRISPR/Cas and a prophage encoding operon were downregulated. Fur binding sites were found upstream of around 20 of the regulated genes. Overall, adaptation to low iron conditions in C. difficile focused on an increase of iron import, a significant replacement of iron requiring metabolic pathways and the restructuring of the cell surface for protection during the complex adaptation phase and was only partly directly regulated by Fur.

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Section: Resultsmentioning

confidence: 99%

Iron Regulation in Clostridioides difficile

et al. 2018

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“…Magnesium ions seemed to have a direct impact on the DA of RhrBR46, without any knowledge of the action of Mg 2+ ions on acetylation. One hypothesis would be to consider the effect of Mg 2+ concentration on the enhancement of acetyltransferase activities, associated with the EPS biosynthesis, as it has been reported for the UDP-N-acetylglucosamine pyrophosphorylase/glucosamine-1-phosphate N-acetyltransferase (GlmU), a bifunctional enzyme catalyzing the synthesis of UDP-N-acetylglucosamine, an essential precursor of cell wall peptidoglycan and lipopolysaccharide synthesis in bacteria [48].…”

Section: Nmr Profiles and Degree Of Acetylationmentioning

confidence: 99%

Description of the Wild Strain Rhizobium rosettiformans DSM26376, Reclassified under Peteryoungia rosettiformans comb.nov., for Producing Glucuronan

et al. 2023

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Glucuronan is a polysaccharide composed of β-(1,4)-linked d-glucuronic acids having intrinsic properties and biological activities recoverable in many fields of application. Currently, the description of Sinorhyzobium meliloti M5N1CS mutant bacterial strain as the sole source of glucuronan makes it relevant to the exploration of new microorganisms producing glucuronan. In this study, the Peteryoungia rosettifformans strain (Rhizobia), was identified as a wild producer of an exopolysaccharide (RhrBR46) related to glucuronan. Structural and biochemical features, using colorimetric assays, Fourier infrared spectroscopy, nuclear magnetic resonance, high pressure size exclusion chromatography coupled to multi-angle light laser scattering, and enzymatic assays allowed the characterization of a polyglucuronic acid, having a molecular mass (Mw¯) of 1.85 × 105 Da, and being partially O-acetylated at C-2 and/or C-3 positions. The concentration of Mg2+ ions in the cultivation medium has been shown to impact the structure of RhrBR46, by reducing drastically its Mw¯ (73%) and increasing its DA (10%). Comparative structural analyses between RhrBR46 and the glucuronan from Sinorhyzobium meliloti M5N1CS strain revealed differences in terms of molecular weight, degree of acetylation (DA), and the distribution of acetylation pattern. These structural divergences of RhrBR46 might contribute to singular properties or biological activities of RhrBR46, offering new perspectives of application.

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“…[23][24][25] The rst step is widespread in bacteria enzymes but is missing in humans, whereas the uridyltransferase step is present in enzymes of both bacteria and humans. 26,27 The acetyltransferase domain could be a suitable target for developing nontoxic antibacterial inhibitors. 28 The study of Ec-GlmU have conrmed that the N-terminal portion of the enzyme contains the uridylyltransferase domain and the C-terminus functions as an acetyltransferase.…”

Section: Introductionmentioning

confidence: 99%