Class Of Siderophores Research Articles

Pyoverdine Analysis - From High-resolution MS/MS Fragmentation to Ion Mobility Measurements.

Microorganisms produce iron chelators called siderophores that are a rich source for drug discovery or plant protective agents. Pyoverdines are a class of siderophores from fluorescent Pseudomonas members and consist of different peptide chains specific to each bacterial species. The structural elucidation and characterization of pyoverdines require comprehensive analytical methods as bacterial extracts are complex mixtures. Here, we present a high-throughput UHPLC-MS/MS pipeline and the application of ion mobility spectrometry to facilitate research in the field of medicine and agriculture.

Chaetomadramines A–E, a class of siderophores with potent neuroprotective activity from the fungus Chaetomium madrasense cib-1

Five hydroxamate siderophores, chaetomadramines A–E (1–5), along with seven known compounds were isolated from the fermented rice culture of the fungus Chaetomium madrasense cib-1. Compounds 1–5 were structurally elucidated on the basis of spectroscopic data, which were a group of unusual hydroxamate siderophores, bearing a long fatty acyl on the α-NH2 of the Nδ-hydroxylated ornithine. Compounds 2–5 were new. The structural elucidation and spectroscopic data of 1 were reported for the first time. Compounds 2–4 significantly improved the survival rates of PC12 cells in the neuroprotective activity assay at the concentration of 40 μM.

Molecular Mechanism of Iron Transport Systems in Vibrio

The ability to acquire iron from the environment is often an important virulence factor for pathogenic bacteria and Vibrios are no exception to this. Vibrios are reported mainly from marine habitats and most of the species are pathogenic. Among those, the pathogenic vibrios eg. V cholerae, V. parahaemolyticus, V. vulnificus causes foodborne illnesses. Vibrios are capable of producing all different classes of siderophores like hydroxamate (aerobactin), catecholate (vibriobactin, fluvibactin), carboxylate (vibrioferrin), and amphiphilic (amphibactin). Every different species of vibrios are capable of utilizing some endogenous or xenosiderophores. Being Gram-negative bacteria, Vibrios import iron siderophore via TonB dependent transport system and unlike other Gamma proteobacteria these usually possess two or even three partially redundant TonB systems for iron siderophore transport. Other than selected few iron siderophores, most pathogenic Vibrios are known to be able to utilize heme as the sole iron source, while some species are capable of importing free iron from the environment. As per the present knowledge, the spectrum of iron compound transport and utilization in Vibrios is better understood than the siderophore biosynthetic capability of individual species.

Siderophore‑antibiotic conjugates: structural diversity and antibacterial activity

Due to uncontrolled growth of antimicrobial resistance, in the near future humanity may return to the «pre-antibiotic era» with no reliable antimicrobial therapy even for previously easily treatable infectious diseases. One of possible solutions is improved delivery of antibiotics to antibiotic-resistant bacterial strains by conjugating them with siderophores (small molecules secreted by microorganisms to absorb essential Fe(III)). The siderophore-modified antibiotic (sideromycin), like a Trojan horse, permeates the bacterial cell as a complex with Fe(III), allowing the antibiotic to reach its biological target. In this review, we describe the structural diversity of siderophore-antibiotic conjugates with the focus on the structure of sideromycin as well as on the relationship between the structure of sideromycin and its antibacterial activity. We analyze main representatives of various classes of siderophores; the structural diversity of sideromycins and their antibacterial activity discussed in detail.

Effect of salinity on the zinc(II) binding efficiency of siderophore functional groups and implications for salinity tolerance mechanisms in barley

Bacteria, fungi and grasses use siderophores to access micronutrients. Hence, the metal binding efficiency of siderophores is directly related to ecosystem productivity. Salinization of natural solutions, linked to climate change induced sea level rise and changing precipitation patterns, is a serious ecological threat. In this study, we investigate the impact of salinization on the zinc(II) binding efficiency of the major siderophore functional groups, namely the catecholate (for bacterial siderophores), α-hydroxycarboxylate (for plant siderophores; phytosiderophores) and hydroxamate (for fungal siderophores) bidentate motifs. Our analysis suggests that the order of increasing susceptibility of siderophore classes to salinity in terms of their zinc(II) chelating ability is: hydroxamate < catecholate < α-hydroxycarboxylate. Based on this ordering, we predict that plant productivity is more sensitive to salinization than either bacterial or fungal productivity. Finally, we show that previously observed increases in phytosiderophore release by barley plants grown under salt stress in a medium without initial micronutrient deficiencies, are in line with the reduced zinc(II) binding efficiency of the α-hydroxycarboxylate ligand and hence important for the salinity tolerance of whole-plant zinc(II) status.

Genetic, structural, and functional diversity of low and high-affinity siderophores in strains of nitrogen fixing Azotobacter chroococcum.

To increase iron (Fe) bioavailability in surface soils, microbes secrete siderophores, chelators with widely varying Fe affinities. Strains of the soil bacterium Azotobacter chroococcum (AC), plant-growth promoting rhizobacteria used as agricultural inoculants, require high Fe concentrations for aerobic respiration and nitrogen fixation. Recently, A. chroococcum str. NCIMB 8003 was shown to synthesize three siderophore classes: (1) vibrioferrin, a low-affinity α-hydroxy carboxylate (pFe = 18.4), (2) amphibactins, high-affinity tris-hydroxamates, and (3) crochelin A, a high-affinity siderophore with mixed Fe-chelating groups (pFe = 23.9). The relevance and specific functions of these siderophores in AC strains remain unclear. We analyzed the genome and siderophores of a second AC strain, A. chroococcum str. B3, and found that it also produces vibrioferrin and amphibactins, but not crochelin A. Genome comparisons indicate that vibrioferrin production is a vertically inherited, conserved strategy for Fe uptake in A. chroococcum and other species of Azotobacter. Amphibactin and crochelin biosynthesis reflects a more complex evolutionary history, shaped by vertical gene transfer, gene gain and loss through recombination at a genomic hotspot. We found conserved patterns of low vs. high-affinity siderophore production across strains: the low-affinity vibrioferrin was produced by mildly Fe limited cultures. As cells became more severely Fe starved, vibrioferrin production decreased in favor of high-affinity amphibactins (str. B3, NCIMB 8003) and crochelin A (str. NCIMB 8003). Our results show the evolution of low and high-affinity siderophore families and conserved patterns for their production in response to Fe bioavailability in a common soil diazotroph.

A Leaf-Inhabiting Endophytic Bacterium, Rhodococcus sp. KB6, Enhances Sweet Potato Resistance to Black Rot Disease Caused by Ceratocystis fimbriata.

Rhodococcus species have become increasingly important owing to their ability to degrade a wide range of toxic chemicals and produce bioactive compounds. Here, we report isolation of the Rhodococcus sp. KB6, which is a new leaf-inhabiting endophytic bacterium that suppresses black rot disease in sweet potato leaves. We determined the 7.0 Mb draft genome sequence of KB6 and have predicted 19 biosynthetic gene clusters for secondary metabolites, including heterobactins, which are a new class of siderophores. Notably, we showed the first internal colonization of host plants with Rhodococcus sp. KB6 and discuss its potential as a biocontrol agent for sustainable agriculture.

Siderophore production of African dust microorganisms over Trinidad and Tobago

Iron (Fe) deposition from African dust has been implicated in a variety of environmental impacts on downwind terrestrial and marine ecosystems throughout the Caribbean. The most abundant form of Fe in African dust is FeIII, which is often not bioavailable. The objective of this study was to determine to what degree microorganisms isolated from African dust collected in Trinidad and Tobago are capable of producing siderophores that mobilize bioavailable Fe into the environment. Aerosol samples were collected for microbial analyses during African dust conditions in the source region (Mali) and downwind sites (Trinidad and Tobago). Microbial community fingerprints, obtained by means of terminal restriction fragment length polymorphism analysis, were compared among aerosol samples and possible Trinidadian sources of locally aerosolized microorganisms (sea water and soils). Ordination of the fingerprint data revealed similarities between aerosols from the source region and the aerosols and soils of downwind regions. Aerosol isolates from the downwind sites were screened for siderophore production using a modified chrome azurol-S (CAS) assay. Twenty-five percent of isolates tested that were sampled under non-dust conditions and 65% of African dust isolates produced at least one type of siderophore; among African dust isolates, all known classes of siderophores were produced. These data support African dust microorganism siderophore production as a viable mechanism by which Fe bioavailability may be increased in downwind locations, given appropriate conditions for microbial proliferation.

Identification and Characterization of a Novel-type Ferric Siderophore Reductase from a Gram-positive Extremophile

Iron limitation is one major constraint of microbial life, and a plethora of microbes use siderophores for high affinity iron acquisition. Because specific enzymes for reductive iron release in gram-positives are not known, we searched Firmicute genomes and found a novel association pattern of putative ferric siderophore reductases and uptake genes. The reductase from the schizokinen-producing alkaliphile Bacillus halodurans was found to cluster with a ferric citrate-hydroxamate uptake system and to catalyze iron release efficiently from Fe[III]-dicitrate, Fe[III]-schizokinen, Fe[III]-aerobactin, and ferrichrome. The gene was hence named fchR for ferric citrate and hydroxamate reductase. The tightly bound [2Fe-2S] cofactor of FchR was identified by UV-visible, EPR, CD spectroscopy, and mass spectrometry. Iron release kinetics were determined with several substrates by using ferredoxin as electron donor. Catalytic efficiencies were strongly enhanced in the presence of an iron-sulfur scaffold protein scavenging the released ferrous iron. Competitive inhibition of FchR was observed with Ga(III)-charged siderophores with K(i) values in the micromolar range. The principal catalytic mechanism was found to couple increasing K(m) and K(D) values of substrate binding with increasing k(cat) values, resulting in high catalytic efficiencies over a wide redox range. Physiologically, a chromosomal fchR deletion led to strongly impaired growth during iron limitation even in the presence of ferric siderophores. Inductively coupled plasma-MS analysis of ΔfchR revealed intracellular iron accumulation, indicating that the ferric substrates were not efficiently metabolized. We further show that FchR can be efficiently inhibited by redox-inert siderophore mimics in vivo, suggesting that substrate-specific ferric siderophore reductases may present future targets for microbial pathogen control.

Borate Binding to Siderophores: Structure and Stability

Well-known as specific iron chelating agents produced by bacteria, it is shown that some, but not all, siderophore classes have an unexpected binding affinity for boron. The relevant criterium is the availability of a vicinal dianionic oxygen containing binding group (i.e., citrate or catecholate). The resulting boron complexes have been characterized by ESI-MS, multinuclear NMR, and DFT calculations. Detailed boron binding constants have been measured for vibrioferrin, rhizoferrin, and petrobactin. The observed affinity of certain siderophores for borate, a common chemical species in the marine but not the terrestrial environment, allows for small, but potentially significant, concentrations of B-siderophores to exist at oceanic pH. We hypothesize that these concentrations could be sufficient for them to function as cell signaling molecules or as mediators of biological boron uptake. In addition, binding of the tetrahedral boron to these siderophores results in a conformation that is different from either the free siderophore or its iron complex and would thus allow a distinction to be made between its iron uptake and any putative cell signaling roles.

Heterobactins: A new class of siderophores from Rhodococcus erythropolis IGTS8 containing both hydroxamate and catecholate donor groups.

We report here on a new class of siderophores isolated from Rhodococcus erythropolis IGTS8, the first structurally characterized from any species of Rhodococcus and for which we suggest the name heterobactins. These siderophores consist of a tripeptide of sequence (N-OH)-L-Orn-Gly-D-Orn-(delta-N-dihydroyxbenzoate). The alpha amino group of the D-Orn is derivatized either as a 2-hydroxybenzoxazolate in heterobactin A or remains free in heterobactin B. The structures were determined by a combination of amino acid analysis, mass spectrometry and NMR methods. The two new compounds are true siderophores in that they relieve iron limited growth in the producing strain. The heterobactins are also transported by other non-producing bacteria. Growth promotion tests using various transport mutants revealed that in E. coli heterobactin A is only recognized by the catecholate receptor Cir while heterobactin B is taken up in both E.coli and A. flavescens JG9 via a hydroxamate transport system.

Coordination Chemistry of the Carboxylate Type Siderophore Rhizoferrin: The Iron(III) Complex and Its Metal Analogs.

Rhizoferrin is a member of a new class of siderophores (microbial iron transport compounds) based on carboxylate and hydroxy donor groups rather than the commonly encountered hydroxamates and catecholates. We have studied the coordination chemistry of rhizoferrin (Rf), as a representative of this group, with Fe(3+), Rh(3+), Cr(3+), Al(3+), Ga(3+), VO(2+), and Cu(2+). The metal complexes have been studied by UV-vis, CD, NMR, and EPR spectroscopies and mass spectrometry. The formation constants for the iron complex have also been measured and yield a log K(LFe) of 25.3. The Rh and Cr rhizoferrin complexes are unusual in that they appear to adopt a chirality about the metal center that is the opposite of the native iron analog. Several of the alternative metal ion complexes are found to have biological activity toward Morganella morganii in a plate type assay.

The specificity of bacterial siderophore receptors probed by bioassays.

The ability to utilize siderophores of bacterial and fungal origin has been studied in wild-type and mutant strains of the enterobacterial genera Salmonella, Escherichia, Shigella, Moellerella, Klebsiella, Enterobacter, Hafnia, Pantoea, Ewingella, Tatumella, Yersinia, and in the non-enterics Aeromonas, Pseudomonas and Aureobacterium. Although only a few representative strains were tested, the results show characteristic genus-specific differences in the utilization of hydroxamate and catecholate siderophores. Moreover, the different response to structural alterations of certain siderophore classes by some wild-type and mutant strains points to variable interacting receptor domains.

Involvement of Bacterial Siderophores in the Remedy of Lime‐induced Chlorosis in Peanut

AbstractA Pseudomonas putida strain isolated from peanut (Arachis hypogaea L.) roots excreted yellow‐green fluorescent siderophores (pigments) when grown under Fe‐deficient conditions. The pigments were purified in their Fe complex form. Chromatography yielded eight peaks, the first two representing &gt;90% of the total. Physico‐chemical characteristics of the materials exhibiting these two peaks were similar to those of the pseudobactin‐pyoverdine class of siderophores. In two growth chamber experiments peanut plants grown on a highly calcareous soil were able to use the Fe from unpurified Fe‐siderophore produced by P. putida cultures. When supplied with 11 mg Fe kg−1 soil as Fe‐siderophore the chlorophyll concentration in the leaves was 75% of that of FeEDDHA (ethylenediamine di‐o‐hydroxyphenylacetic acid)‐treated plants. Control plants that were not fertilized with Fe contained only 35% chlorophyll of that of FeEDDHA‐treated plants. Nonferrated siderophores were ineffective in correcting Fe deficiency. Relative chlorophyll concentration in the leaves of the treatment supplemented with Fe‐siderophore and bacterial cells was similar to that of applications that did not contain bacterial cells. In a second experiment, 100% remedy of the deficiency was achieved when 19 mg Fe kg−1 soil as Fe‐siderophores was applied in comparison to 1.2 mg Fe kg−1 soil as FeEDDHA. The degree of remedy of chlorosis was independent on the frequency of application (two or four times a month). In these treatments total bacterial and fluorescent pseudomonad counts [colony‐forming units (cfu) g−1 dry roots] were significantly higher than in the unfertilized and FeEDDHA treatments. The level of DTPA (diethylenetriam‐inepentaacetic acid) extractable Fe at the end of the growth period in both experiments was significantly higher in Fe‐siderophore amended soils than in FeEDDHA‐treated soils. In a third experiment, the siderophore Ferrioxamine B (FOB) was used as a source of Fe. Iron applications of 22.5 and 45 mg Fe kg−1 soil as FOB were required to reach 70 and 97%, of the chlorophyll concentration of the FeEDDHA‐treated plants, respectively. It was concluded that bacterial siderophores may serve as a remedy to lime‐induced chlorosis in plants grown in calcareous soils.

Transport properties of N-acyl derivatives of the coprogen and ferrichrysin classes of siderophores inNeurospora crassa

Several derivatives of the coprogen and ferrichrysin classes of siderophores were synthesized as potential affinity labels of the iron uptake system inNeurospora crassa. While only one of these compounds has proved useful as an affinity label, all were recognized and transported byNeurospora crassa. One derivative, chloroacetyl-ferrichrysin, proved to be an unexpectedly potent reversible inhibitor (K1=0.4 μM) of both ferrichrysin and coprogen uptake, similar to the natural siderophore, ferrirubin. The reported results provide further understanding of the steric and electronic requirements of siderophores for the iron uptake system inNeurospora crassa.