Chemical Synthesis of Staphyloferrin B Affords Insight into the Molecular Structure, Iron Chelation, and Biological Activity of a Polycarboxylate Siderophore Deployed by the Human Pathogen

Staphyloferrin B (SB) is a citrate-based polycarboxylate siderophore produced and utilized by the human pathogen Staphylococcus aureus for acquiring iron when colonizing the vertebrate host. The first chemical synthesis of SB is reported, which enables further molecular and biological characterization and provides access to structural analogues of the siderophore. Under conditions of iron limitation, addition of synthetic SB to bacterial growth medium recovered the growth of the antibiotic resistant community isolate S. aureus USA300 JE2. Two structural analogues of SB, epiSB and SBimide, were also synthesized and employed to investigate how epimerization of the citric acid moiety or imide formation influence its function as a siderophore. Epimerization of the citric acid stereocenter perturbed the iron-binding properties and siderophore function of SB as evidenced by experimental and computational modeling studies. Although epiSB provided growth recovery to S. aureus USA300 JE2 cultured in iron-deficient medium, the effect was attenuated relative to that of SB. Moreover, SB more effectively sequestered the Fe(III) bound to human holo-transferrin, an iron source of S. aureus, than epiSB. SBimide is an imide analogous to the imide forms of other citric acid siderophores that are often observed when these molecules are isolated from natural sources. Here, SBimide is shown to be unstable, converting to native SB at physiological pH. SB is considered to be a virulence factor of S. aureus, a pathogen that poses a particular threat to public health because of the number of drug-resistant strains emerging in hospital and community settings. Iron acquisition by S. aureus is important for its ability to colonize the human host and cause disease, and new chemical insights into the structure and function of SB will inform the search for new therapeutic strategies for combating S. aureus infections.Alfred Benzon Foundation (Postdoctoral fellowship)Pacific Southwest Regional Center of ExcellenceAlfred P. Sloan Foundatio

Hydrogen release through catalyzed methanolysis of solid sodium borohydride

Hannauer, J. Demirci, U. B. Pastor, G. Geantet, C. Herrmann, J. M. Miele, P.International audienceMethanolysis of sodium borohydride (NaBH4) is a way of recovering the hydrogen stored in the hydride. Though the reaction is spontaneous, it can be accelerated by virtue of Co-TiO2 or Ru-TiO2 catalysts. Under our experimental conditions, Co-TiO2 shows high catalytic performances, higher than those of Ru-TiO2. Hydrogen generation rates of 144 to 644 L(H-2) min(-1) g(-1)(Co) were measured as the Co content was decreased. The kinetic parameters of the catalyzed reaction were determined. The Co-TiO2- catalyzed methanolysis follows a power law, i.e. r(20) k center dot[NaBH4](1.3)center dot[CH3OH](0.9) with k = 1.8 x 10(-2) s(-1). The Langmuir-Hinshelwood bimolecular mechanism accounts for the kinetics. The apparent activation energy was found to be 20.4 kJ mol(-1) whereas that of the catalyzed hydrolysis was 49.4 kJ mol(-1). Indeed, the catalyzed methanolysis was compared to the catalyzed hydrolysis as well as the catalyzed ethanolysis. For instance, it was remarked that water in methanol has a detrimental effect on the H-2 release kinetics. In parallel, the gravimetric hydrogen density of the system NaBH4-CH3OH has been optimized. Under our experimental conditions, it was found that the highest capacity that can be achieved is 3.4 wt%

Review of fragmentation of synthetic single-stranded oligonucleotides by tandem mass spectrometry from 2014 to 2022

The fragmentation of oligonucleotides by mass spectrometry allows for the determination of their sequences. It is necessary to understand how oligonucleotides dissociate in the gas phase, which allows interpretation of data to obtain sequence information. Since 2014, a range of fragmentation mechanisms, including a novel internal rearrangement, have been proposed using different ion dissociation techniques. The recent publications have focused on the fragmentation of modified oligonucleotides such as locked nucleic acids, modified nucleobases (methylated, spacer, nebularine and aminopurine) and modification to the carbon 2′-position on the sugar ring; these modified oligonucleotides are of great interest as therapeutics. Comparisons of different dissociation techniques have been reported, including novel approaches such as plasma electron detachment dissociation and radical transfer dissociation. This review covers the period 2014–2022 and details the new knowledge gained with respect to oligonucleotide dissociation using tandem mass spectrometry (without priori sample digestion) during that time, with a specific focus on synthetic single-stranded oligonucleotides

Advancements in the characterisation of oligonucleotides by high performance liquid chromatography-mass spectrometry in 2021: A short review

The first oligonucleotide therapeutic was approved by the Food and Drug Administration in 1998, and since then, 12 nucleic acids have been commercialised as medicines. To be approved, the oligonucleotides need to be identified and characterised as well as its related impurities. Different methods exist, but the most commonly used is ion-pairing reversed-phase liquid chromatography with tandem mass spectrometry. The separation obtained depends on the mobile phase and column used. Other methods have been developed, notably by using hydrophilic interaction chromatography and two-dimensional high performance liquid chromatography. Furthermore, ion-pairing reversed-phase high performance liquid chromatography ultra-violet spectroscopy detection and mass spectrometry has been optimised for the analysis of methylated nucleobases due to the utilisation of this modification in the drugs. This review covers the recent advancements in the analysis and characterisation of oligonucleotides in 2021 by high performance liquid chromatography mass spectrometry, notably by hydrophilic interaction chromatography and two-dimensional liquid chromatography but also the different parameters that influence the analysis by ion-pairing reversed-phase high performance liquid chromatography, the characterisation of methylated nucleobases, and the recent software developed for oligonucleotides

Biosynthesis of the pyoverdine siderophore of Pseudomonas aeruginosa involves precursors with a myristic or a myristoleic acid chain

Pyoverdine I (PVDI) is the major siderophore produced by Pseudomonas aeruginosa to import iron. Biosynthesis of this chelator involves non‐ribosomal peptide synthetases and other enzymes. PvdQ is a periplasmic enzyme from the NTN hydrolase family and is involved in the final steps of PVDI biosynthesis. A pvdQ mutant produces two non‐fluorescent PVDI precursors with a higher molecular mass than PVDI. In the present study, we describe the use of mass spectrometry to determine the structure of these PVDI precursors and show that they both contain a unformed chromophore like ferribactin, and either a myristic or myristoleic chain that must be removed before PVDI is secreted into the extracellular medium

Hydrogen release through catalyzed methanolysis of solid sodium borohydride

Hannauer, J. Demirci, U. B. Pastor, G. Geantet, C. Herrmann, J. M. Miele, P.International audienceMethanolysis of sodium borohydride (NaBH4) is a way of recovering the hydrogen stored in the hydride. Though the reaction is spontaneous, it can be accelerated by virtue of Co-TiO2 or Ru-TiO2 catalysts. Under our experimental conditions, Co-TiO2 shows high catalytic performances, higher than those of Ru-TiO2. Hydrogen generation rates of 144 to 644 L(H-2) min(-1) g(-1)(Co) were measured as the Co content was decreased. The kinetic parameters of the catalyzed reaction were determined. The Co-TiO2- catalyzed methanolysis follows a power law, i.e. r(20) k center dot[NaBH4](1.3)center dot[CH3OH](0.9) with k = 1.8 x 10(-2) s(-1). The Langmuir-Hinshelwood bimolecular mechanism accounts for the kinetics. The apparent activation energy was found to be 20.4 kJ mol(-1) whereas that of the catalyzed hydrolysis was 49.4 kJ mol(-1). Indeed, the catalyzed methanolysis was compared to the catalyzed hydrolysis as well as the catalyzed ethanolysis. For instance, it was remarked that water in methanol has a detrimental effect on the H-2 release kinetics. In parallel, the gravimetric hydrogen density of the system NaBH4-CH3OH has been optimized. Under our experimental conditions, it was found that the highest capacity that can be achieved is 3.4 wt%