Reductive Dissolution of Jarosite by Acidiphilium cryptum in Presence of Chelating Agents and Dissolved Iron

The mutual effects of Acidiphilium cryptum, nine ligands and dissolved iron on jarosite dissolution were studied in a 2³ full factorial experiment. Ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA) and oxalic acid were able to dissolve jarosite chemically, but bacterial action enhanced this dissolution; from 88 ± 1, 46 ± 0 and 36 ± 0 µmol/L d to 129 ± 3, 177 ± 5 and 106 ± 14 µmol/L d, respectively. Oxalate and NTA exhibited a synergistic effect with dissolved iron and bacteria that may be explained by a model in which iron complexes behave as redox mediators in the microbial respiration in which jarosite, a solid substrate, serves as terminal electron acceptor.     

Long-Term Waterlogging: Nutrient,Gas Exchange,Photochemical and Pigment Characteristics of Eucalyptus nitens Saplings

Imposition of waterlogging for eight months induced morphological adaptation in Eucalyptus nitens in the form of adventitious and aerenchymatous roots and hypertrophy of stems. Foliar calcium (1.3-fold), potassium (2-fold) and phosphorus (2.4-fold) were lower and iron (5.6-fold) was higher in waterlogged than control saplings. Stem Ca (1.7-fold) was lower, whereas Mn (1.8-fold) and Fe (117-fold) were higher in waterlogged than control saplings. Distinct purple pigmentation was observed in xylem tissues of waterlogged saplings. A significant reduction in the maximum photosynthetic rate and photochemical efficiency was observed in waterlogged compared to control saplings. Although chlorophyll levels were similar, the xanthophyll cycle pool size was significantly greater in waterlogged saplings and may have contributed to the relatively greater capacity for light energy dissipation observed. Predawn xanthophyll cycle engagement was significantly greater in waterlogged than control saplings, despite relatively mild temperatures. Foliar anthocyanin concentration was higher in waterlogged than control saplings.     

Removal of heavy metals from water effluents using supermacroporous metal chelating cryogels

Applications of IDA in, for example, immobilized metal ion affinity chromatography for purification of His-tagged proteins are well recognized. The use of IDA as an efficient chelating adsorbent for environmental separations, that is, for the capture of heavy metals, is not studied. Adsorbents based on supermacroporous gels (cryogels) bearing metal chelating functionalities (IDA residues and ligand derived from derivatization of epoxy-cryogel with tris(2-aminoethyl)amine followed by the treatment with bromoacetic acid (defined as TBA ligand)) have been prepared and evaluated on capture of heavy metal ions. The cryogels were prepared in plastic carriers, resulting in desired mechanical stability and named as macroporous gel particles (MGPs). Sorption and desorption experiments for different metals (Cu2+, Zn2+, Cd2+, and Ni2+ with IDA adsorbent and Cu2+ and Zn2+ with TBA adsorbent) were carried out in batch and monolithic modes, respectively. Obtained capacities with Cu2+ were 74 μmol/mL (TBA) and 19 μmol/mL gel (IDA). The metal removal was higher for pH values between pH 3 and 5. Both adsorbents showed improved sorption at lower temperatures (10°C) than at higher (40°C) and the adsorption significantly dropped for the TBA adsorbent and Zn2+ at 40°C. Desorption of Cu2+ by using 1 M HCl and 0.1 M EDTA was successful for the IDA adsorbent whereas the desorption with the TBA adsorbent needs further attention. The result of this work has demonstrated that MGPs are potential treatment alternatives within the field of environmental separations and the removal of heavy metals from water effluents.     

Isolation and identification of hydroxamate siderophores of ericoid mycorrhizal fungi

Three ericoid mycorrhizal fungi were grown in pure culture under iron deprivation: (i) the ascomyceteHymenoscyphus ericae, a characteristic endophyte of ericaceous plants on acid soils; (ii) the hyphomyceteOidiodendron griseum, an ericoid mycorrhizal fungus which is also a soil-borne fungus able to colonize wood; and (iii) an endophyte of the calciculous ericaceous plantRhodothamnus chamaecistus. All three fungi produced several hydroxamate siderophores which were isolated in the ferric form by adsorption to Amberlite XAD-2, gel chromatography on Sephadex LH20 and by HPLC on a C₁₈ reversed-phase column. Siderophores were identified by (i) co-chromatography with known fungal siderophores, (ii) ion spray mass spectrometry after semi-preparative HPLC and (iii) analyzing their electrophoretic behavior. WhileH. ericae andO. griseum were similar in producing ferricrocin as their principal siderophore, the endophyte ofR. chamaecistus produced mainly fusigen.     

Protein selectivity in immobilized metal affinity chromatography based on the surface accessibility of aspartic and glutamic acid residues

The interaction of different species variants of cytochrome c and myoglobin, as well as hen egg white lysozyme, with the hard Lewis metal ions Al³⁺, Ca²⁺, Fe³⁺, and Yb³⁺ and the borderline metal ion Cu²⁺, immobilized to iminodiacetic acid (IDA)-Sepharose CL-4B, has been investigated over the rangepH 5.5–8.0. With appropriately chosen buffer and metal ion conditions, these proteins can be bound to the immobilized M ⁿ +-IDA adsorbents via negatively charged amino acid residues accessible on the protein surface. For example, tuna heart cytochrome c, which lacks surface-accessible histidine residues, readily bound to the Fe³⁺-IDA adsorbent, while the other proteins also showed affinity toward immobilized Fe³⁺-IDA adsorbents when buffers containing 30 mM of imidazole were used. These studies document that protein selectivity can be achieved with hard-metalion immobilized metal ion affinity chromatography (IMAC) systems through the interaction of surfaceexposed aspartic and glutamic acid residues on the protein with the immobilized M ⁿ +-IDA complex. These investigations have also documented that the so-called soft or borderline immobilized metal ions such as the Cu²⁺-IDA adsorbent can also interact with surface-accessible aspartic and glutamic acid residues in a protein-dependent manner. A relationship is evident between the number and extent of clustering of the surfaceaccessible aspartic and glutamic acid residues and protein selectivity with these IMAC systems. The use of elution buffers which contain organic compound modifiers which replicate the carboxyl group moieties of these amino acids on the surface of proteins is also described.Abbreviations IDA iminodiacetic acid - IDA-Mⁿ⁺ iminodiacetic acid chelated to metal ion - IMAC immobilized metal affinity chromatography - DHCC dog heart cytochrome c - HHCC horse heart cytochrome c, THCC, tuna heart cytochrome c - HMYO horse skeletal muscle myoglobin - SMYO sheep skeletal muscle myoglobin - HEWL hen egg white lysozyme     

The role of free amino groups of peptides and proteins in the Folin-Lowry and biuret methods

On an equal weight basis polymyxin B and EM 49 which do not contain tyrosine or tryptophan yielded the same colour intensity as proteins in the Folin-Lowry and biuret methods. But, in the absence of reagent C (alkaline copper reagent) polymyxin B and EM 49 yielded no colour in the Folin-Lowry method. Mono-, di- and tri-formyl polymyxins B formed identical amounts of coloured complexes as polymyxin B in the two methods. However, the tetra- and penta-formyl polymyxins B yielded only one-fifth and one-sixth, respectively, of the expected colour in the Folin-Lowry method. Similarly, 40% and 30%, respectively, of the anticipated amount of colour is formed in the biuret method. Formylated and methylated lysozyme and bovine serum albumins form only 70–75% of the expected colour in the Folin-Lowry method. Since formation of colour by reduction of Folin reagent, in the Folin-Lowry method, is at least partly due to complexes of copper, it was inferred that polymyxin B as well as its mono-, di- and tri-formyl derivatives on the one hand and the tetra- and penta-formyl derivatives on the other differ in their ability to complex Cu(II) The former group of compounds was indeed found to complex as many as three Cu(II) ions whereas the tetra- and penta-formyl polymyxins B complexed only one equivalent, under conditions of excess Cu(II). Under conditions of low Cu(II), polymyxin B and all its derivatives complexed only one Cu(II). In proteins, sites other than amino groups which complex Cu(II) probably play a major role in the reduction of the Folin reagent, since methylated lysozyme and bovine serum albumin yield 70–75% of the colour formed by the unmodified proteins in the Folin-Lowry reaction.     

Responses of nitrogen-fixing and nitrate-supplied alfalfa (Medicago sativa L.) to iron chelates in an alkaline hydroponic medium

Ferric ethylenediamine di-(o-hydroxyphenylacetate) (FeEDDHA) and ferric hydroxyethylethylenediaminetriacetic acid (FeHEDTA) were evaluated as Fe sources for hydroponic growth of alfalfa (Medicago sativa L., cv. Mesilla), either dependent on N₂ fixation or supplied with NO₃. The hydroponic medium was maintained at pH 7.5 by addition of CaCO₃. Nitrogen-fixing cultures were inoculated with Rhizobium meliloti 102 F51 and grown in medium without added nitrogen. After five to seven weeks of growth under greenhouse conditions, plants were harvested. Nitrogen fixation was measured by the acetylene reduction method.When FeEDDHA was supplied, growth of alfalfa, whether dependent on N₂ fixation or supplied with NO₃, was severely limited at concentrations typically used in hydroponic medium (10 or 20 M). Maximum yield of NO₃-supplied alfalfa was obtained at 100 M while maximum yield of N₂-fixing alfalfa was obtained in the range of 33 to 200 M FeEDDHA. Nodule fresh weights and N₂ fixation rates increased with FeEDDHA concentration up to 33 M and remained essentially constant up to 200 M. With FeHEDTA, maximum yields of both NO₃-grown and N₂-fixing alfalfa were obtained at 10 M. Growth of NO₃-supplied plants was inhibited at 200 M FeHEDTA while growth of N₂-fixing plants was inhibited at 100 M FeHEDTA. The numbers of nodules per plant increased between 3.3 and 10 M FeHEDTA; however, inhibition of nodule formation occurred at a concentration of 33 M or higher. Nodule weights per plant and N₂ fixation rates were depressed at 3.3 M as well as at 100 M FeHEDTA. The results suggest that alfalfa dependent on N₂ fixation is more sensitive to limited Fe availability than alfalfa supplied with NO₃.     

A possible role for copper-mediated oxidation of thiols in the regulation of the release of luteinizing hormone releasing hormone from isolated hypothalamic granules

Abstract: A role for copper in the release of luteinizing hormone releasing hormone (LHRH) from hypothalamic neurons has been previously proposed. To elucidate further the mechanism of action of copper, we addressed two questions: (a) what is the active form of copper that interacts with the LHRH granule (ionic or chelated)? and (b) is copper-stimulated LHRH release a result of an interaction of copper with thiol groups and, if so, does it require oxygen? Granules were isolated from hypothalami of adult male rats and were then incubated at 37°C for 3–5 min in a buffered medium. When granules were incubated with various copper complexes, CuATP stimulated LHRH release by 45 ± 4% (mean ± SE), copper tartrate by 44 ± 4%, CuBSA by 27 ± 7%, and copper histidine by 16 ± 6%. Neither CuEDTA nor CuCl₂ stimulated LHRH release. CuATP-stimulated LHRH release from granules incubated under N₂ was 50% of that incubated under air. Furthermore, the CuATP-stimulated release of LHRH was completely inhibited by dithiothreitol or glutathione (10^?3M each), partially (40–50%) by iodoacetate or 5,5-dithiobis-(2-nitrobenzoic acid), and not at all by oxidized dithiothreitol. Thus, chelated copper, rather than ionic copper, is the active form of the metal, and the action of copper involves an oxidation reaction and granule thiol groups. The precise mechanism of action of copper, however, has yet to be elucidated. We propose that copper may affect LHRH release as follows: copper, bound to an intracellular chelator (protein, peptide, or amino acid), oxidizes thiols of the LHRH granule, leading to a change in granule-membrane permeability and hence to LHRH release.     

Structural studies in solution and in the solid state on the zinc chelate of 2-hydroxy-(4-methylthio)butanoic acid, an effective mineral supplement in animal feeding

The bis-chelate complex of Zn²⁺ with 2-hydroxy-(4-methylthio)butanoate (MHA_-H the anion derived from the so-called methionine hydroxy-analogue, MHA) is an effective, bioavailable mineral supplement for animal feeding. It can be obtained in two solid forms: the anhydrous [Zn(OC(O)CH₂CH(OH)CH₂CH₂SCH₃)₂] and the corresponding dihydrate species, both well distinguishable by IR spectroscopy and powder X-ray diffraction. The crystal and molecular structure of the dihydrate form has been solved by single-crystal X-ray diffraction. It consists of dinuclear bis-chelate species with a bridging carboxylate group, both zinc atoms displaying hexacoordination involving all the hydroxyl and carboxyl groups from the four MHA_-H anions and three oxygens from different water molecules. The fourth water molecule does not participate in coordination. Therefore, the dihydrate complex must be formulated as [Zn₂(OC(O)CH₂CH(OH)CH₂CH₂SCH₃)₄(H₂O)₃] · H₂O (1). A molecular computational analysis has been carried out by density functional theory (DFT) on three possible MHA_-H zinc chelates, i.e. the dinuclear bis-chelate observed in the solid state, the mononuclear bis-chelate diaquo-complex, and the monochelate tetraaquo-complex. Calculations have suggested that between the dinuclear and mononuclear bis-chelates, the preferred form in aqueous solution may be the second one. Moreover, both ¹H (chemical shifts and relaxation rates) and ¹³C NMR data provide further evidence for the formation of Zn/MHA_-H chelates in solution.     

Study of the interaction between a histidine-deoxyguanosine hybrid and cisplatin

 A histidine-2′-deoxyguanosine hybrid, Phac-Hse(p^5′dG)-His-OH (I), was synthesized, and its reaction with cisplatin was monitored by reversed-phase HPLC and ¹H NMR. Two new compounds, II and III, were observed to be simultaneously formed, II in larger proportion than III. These products were isolated after HPLC purification and extensively characterized. Both II and III contained platinum, had the same mass and showed a bathochromic displacement of their absorption maxima with respect to that of I. Both remained undegraded upon enzymatic digestion and yielded I when treated with NaCN. From these data and the information provided by ¹H NMR analysis, we inferred that II and III were constitutional isomers, in particular chelates in which platinum was coordinated to the N7 of guanine and either the N^π or the N^τ imidazole nitrogens, respectively. No preference of the metal for either of these N-donors was observed. Received: 3 May 1999 / Accepted: 24 August 1999