Joanna C. Renshaw

Fungal siderophores: structures, functions and applications

Siderophores are low molecular weight, iron-chelating ligands produced by nearly all microorganisms. Fungi synthesize a wide range of hydroxamate siderophores. This review considers the chemical and biological aspects of these siderophores, their distribution amongst fungal genera and their possible applications. Siderophores function primarily as iron transport compounds. Expression of siderophore biosynthesis and the uptake systems is regulated by internal iron concentrations. Transport of siderophores is an energy-dependent process and is stereoselective, depending on recognition of the metal ion coordination geometry. In addition to transporting iron, siderophores have other functions and effects, including enhancing pathogenicity, acting as intracellular iron storage compounds and suppressing growth of other microorganisms. Siderophores can complex other metals apart from iron, in particular the actinides. Because of their metal-binding ability there are potential applications for siderophores in medicine, reprocessing of nuclear fuel, remediation of metal-contaminated sites and the treatment of industrial waste.

Interactions of silver nanoparticles with Pseudomonas putida biofilms.

Silver nanoparticles (Ag NPs) may present a risk to the environment due to their expected toxicity and wide exposure. The interactions between Ag NPs and laboratory-grown Pseudomonas putida biofilms were investigated under a range of environmentally relevant conditions (pH 6 and 7.5; presence and absence of Suwannee River fulvic acid (SRFA)) over 4 days. In the absence of SRFA, there was extensive sloughing of the biofilm bacteria into suspension implying NP-bacterial interactions and potential effects on NP transport in the environment. In the presence of SRFA, sloughing of cells into suspension was reduced under some conditions and Ag NPs and their aggregates were observed and quantified on and in the bacterial cells in the biofilm. Viability of the cells in all cases appear unchanged by the presence of Ag NPs. Cell viability was independent of the concentration of NPs in solution, but sloughing rates varied substantially, sometimes in a dose-dependent manner. The results suggest that biofilms are impacted by Ag NPs when SRFA was not present, and that SRFA increases uptake and bioaccumulation of Ag NPs to biofilms, perhaps resulting in longer term effects, which need further investigation.

Formation of Nanoscale Elemental Silver Particles via Enzymatic Reduction by Geobacter sulfurreducens

ABSTRACT Geobacter sulfurreducens reduced Ag(I) (as insoluble AgCl or Ag+ ions), via a mechanism involving c-type cytochromes, precipitating extracellular nanoscale Ag(0). These results extend the range of metals known to be reduced by Geobacter species and offer a method for recovering silver from contaminated water as potentially useful silver nanoparticles.

Impact of silver nanoparticles on natural marine biofilm bacteria

There has been a recent increase in the use of silver nanoparticles (Ag NPs) in a wide range of consumer products due to their highly effective antimicrobial properties. However, Ag NPs give cause for concern since their wide use makes them likely to be released into aquatic ecosystems and potentially affect natural bacterial communities. In this study marine biofilms were grown in situ in a coastal site (Singapore Harbour) and exposed in the laboratory for a further 24h to 0-2000 μg L⁻¹ of well characterised Ag NPs. Increasing concentrations of Ag NPs caused a significant decrease in biofilm volume and biomass, and Ag uptake by biofilms per unit of volume was also dependent on concentration. Terminal fragment length polymorphisms and subsequent cluster and phylogenetic analysis showed the presence of major bacterial groups in biofilms irrespective of treatment with Ag NPs. This implies that even at the highest concentrations studied these taxonomic groups were not displaced. Nevertheless, biofilm succession was impeded on Ag NP treated biofilms, affecting the relative abundance of major bacterial groups in the biofilm community, with potential longer term effects on biofilm development and function.

Reduction of uranium(VI) phosphate during growth of the thermophilic bacterium thermoterrabacterium ferrireducens

ABSTRACT The thermophilic, gram-positive bacterium Thermoterrabacterium ferrireducens coupled organotrophic growth to the reduction of sparingly soluble U(VI) phosphate. X-ray powder diffraction and X-ray absorption spectroscopy analysis identified the electron acceptor in a defined medium as U(VI) phosphate [uramphite; (NH4)(UO2)(PO4) · 3H2O], while the U(IV)-containing precipitate formed during bacterial growth was identified as ningyoite [CaU(PO4)2 · H2O]. This is the first report of microbial reduction of a largely insoluble U(VI) compound.

Uptake of Sr 2+ and Co 2+ into biogenic hydroxyapatite: implications for biomineral ion exchange synthesis

Biomineral hydroxyapatite (Bio-HAp) produced by Serratia sp. has the potential to be a suitable material for the remediation of metal contaminated waters and as a radionuclide waste storage material. Varying the Bio-HAp manufacturing method was found to influence hydroxyapatite (HAp) properties and consequently the uptake of Sr(2+) and Co(2+). All the Bio-HAp tested in this study were more efficient than the commercially available hydroxyapatite (Com-HAp) for Sr(2+) and Co(2+) uptake. For Bio-HAp the uptake for Sr(2+) and Co(2+) ranged from 24 to 39 and 29 to 78 mmol per 100 g, respectively. Whereas, the uptake of Sr(2+) and Co(2+) by Com-HAp ranged from 3 to 11 and 4 to 18 mmol per 100 g, respectively. Properties that increased metal uptake were smaller crystallite size (<40 nm) and higher surface area (>70 m(2) g(-1)). Organic content which influences the structure (e.g., crystallite arrangement, size and surface area) and composition of Bio-HAp was also found to be important in Sr(2+) and Co(2+) uptake. Overall, Bio-HAp shows promise for the remediation of aqueous metal waste especially since Bio-HAp can be synthesized for optimal metal uptake properties.

Development and Application of an Assay for Uranyl Complexation by Fungal Metabolites, Including Siderophores

ABSTRACT An assay to detect UO22+ complexation was developed based on the chrome azurol S (CAS) assay for siderophores (B. Schwyn and J. B. Neilands, Anal. Biochem. 160:47-56, 1987) and was used to investigate the ability of fungal metabolites to complex actinides. In this assay the discoloration of two dyed agars (one containing a CAS-Fe3+ dye and the other containing a CAS-UO22+ dye) caused by ligands was quantified. The assay was tested by using the siderophore desferrioxamine B (DFO), and the results showed that there was a regular, reproducible relationship between discoloration and the amount of siderophore added. The ratio of the discoloration on the CAS-UO22+ agar to the discoloration on the CAS-Fe3+ agar was independent of the amount of siderophore added. A total of 113 fungi and yeasts were isolated from three soil samples taken from the Peak District National Park. The fungi were screened for the production of UO22+ chelators by using the CAS-based assay and were also tested specifically for hydroxamate siderophore production by using the hydroxamate siderophore auxotroph Aureobacterium flavescens JG-9. This organism is highly sensitive to the presence of hydroxamate siderophores. However, the CAS-based assay was found to be less sensitive than the A. flavescens JG-9 assay. No significant difference between the results for each site for the two tests was found. Three isolates were selected for further study and were identified as two Pencillium species and a Mucor species. Our results show that the new assay can be effectively used to screen fungi for the production of UO22+ chelating ligands. We suggest that hydroxamate siderophores can be produced by mucoraceous fungi.

Bacterially produced calcium phosphate nanobiominerals : sorption capacity, site preferences, and stability of captured radionuclides

A Serratia sp. bacterium manufactures amorphous calcium phosphate nanominerals (BHAP); this material has shown increased sorption capacity for divalent radionuclide capture. When heat-treated (≥450 °C) the cell biomass is removed and the biominerals are transformed to hydroxyapatite (HAP). Using a multimethod approach, we have elucidated both the site preferences and stability of analogue radionuclide incorporation for Sr, Co, Eu, and U. Strontium incorporates within the bulk amorphous inorganic phase of BHAP; however, once temperature modified to crystalline HAP, bonding was consistent with Sr substitution at the Ca(1) and/or Ca(2) sites. Cobalt incorporation occurs within the bulk inorganic amorphous phase of BHAP and within the amorphous grain boundaries of HAP. Europium (an analogue for trivalent actinides) substituted at the Ca(2) and/or the Ca(3) position of tricalcium phosphate, a known component of HAP grain boundaries. Uranium was surface complexed with no secondary minerals detected. With multiple sites for targeted radionuclide incorporation, high loadings, and good stability against remobilization, BHAP is shown to be a potential material for the remediation of aqueous radionuclide in groundwater.

A new incorporation mechanism for trivalent actinides into bioapatite : a TRLFS and EXAFS study

One of the most toxic byproducts of nuclear power and weapons production is the transuranics, which have a high radiotoxicity and long biological half-life due to their tendency to accumulate in the skeletal system. This accumulation is inhomogeneous and has been associated with the chemical properties and structure of the bone material rather than its location or function. This suggests a chemical driving force to incorporation and requires an atomic scale mechanistic understanding of the incorporation process. Here we propose a new incorporation mechanism for trivalent actinides and lanthanides into synthetic and biologically produced hydroxyapatite. Time-resolved laser fluorescence spectroscopy and extended X-ray absorption fine structure have been used to demonstrate that trivalent actinides and lanthanides incorporate into the amorphous grain boundaries of apatite. This incorporation site can be used to explain patterns in uptake and distribution of radionuclides in the mammalian skeletal system.

Rhodotorulic acid production by Rhodotorula mucilaginosa

Rhodotorula mucilaginosa produces the siderophore rhodotorulic acid (RA) when grown in iron-limited conditions. R. mucilaginosa grew at rates between 0.10 and 0.19 h(-1) in iron-restricted conditions, depending on the carbon source, and at 0.23 h(-1) in iron-sufficient conditions. In bioreactors inoculated with iron-starved pre-cultures, initial specific growth rates in batch culture were dependent on the iron concentration. The critical dilution rate (Dcrit, at which steady state cultures cannot be sustained) in continuous cultures was also dependent on the iron concentration and was lower than mu(max) in batch culture. Sucrose was the best carbon source for RA production [287+/-11 micromol (g biomass)(-1)] and production could be further increased by supplementing the medium with the precursors acetate [460+/-13 micromol (g biomass)(-1)], ornithine [376+/-6 micromol (g biomass)(-1)], or both [539+/-15 micromol (g biomass)(-1)]. Citric acid was an effective suppresser of RA production. RA was produced in a growth rate dependent manner and was optimally produced at pH 6.5.