Removal of iron and formation of copper(i) from solutions containing iron(ii) and copper(ii) by addition of chloride ion or acetonitrile

Solutions of Cu(II) and Fe(II) establish the redox equilibrium

$\text{Cu(II) + Fe(II)}\text{?}\text{K}\text{Cu(I) + Fe(III)}$

which is displaced to the right by addition of either Cl^? or acetonitrile (AN). Log K varies from ?10.5 in water to about ?2.5 in 4 M NaCl or AN, allowing iron to be removed selectively from copper (II) solutions either by solvent extraction with Versatic acid or by precipitation as goethite or j jarosite. To establish the required conditions Eh-pH diagrams have been developed for the CuH₂OCl and CuH₂OANSO₄²-systems at 25°C and 90°C. It is demonstrated that the catalytic effect of Cu(II) on the oxidation of Fe(II) to Fe(III) by O₂ is dependent on the concentration of Cl^? or AN and on the position of this redox equilibrium. Applications to removing iron from hydrometallurgical solutions are discussed and tested.     

Cuprous hydrometallurgy Part IV. Rates and equilibria in the reaction of copper sulphides with copper(II) sulphate in aqueous acetonitrile

Copper(II) sulphate in solutions of aqueous acetonitrile leaches copper from copper sulphides to form stable copper(I) sulphate solutions. Covellite and chalcopyrite are oxidised and leached more rapidly in the early stages of leaching with acidic CuSO₄/CH₃CN/H₂O than with acidic iron(III) sulphate in water. A redox equilibrium between copper(I) sulphate, copper(II) sulphate and partially leached solid copper sulphide, Cu_xS, is established. The equilibrium concentration of Cu₂SO₄ and the value of x in Cu_xS, in solutions saturated with CuSO₄, are interdependent if the acetonitrile concentration is constant. This behaviour is considered in terms of the structural and electrochemical changes which occur, in the solids Cu₂S and CuS, as leaching proceeds. According to the activities of acetonitrile, of copper(I) sulphate and of copper(II) sulphate, i.e. according to the redox potential of the solution, CuS either can be oxidised by copper(II) sulphate to a less copper-rich copper sulphide and even to sulphur, or reduced by copper(I) sulphate to a more copper-rich sulphide, up to Cu₂S, in acidic solutions containing CuSO₄, Cu₂SO₄, CH₃CN and water. This observation leads to an easy method of generating Cu₂S from CuS or from sulphur.     

Resonance Raman spectra of iron(III)-, copper(II)-, cobalt(III)-, and manganese(III)-transferrins and of bis(2,4,6-trichlorophenolato)diimidazolecopper(II) monohydrate, a possible model for copper(II) binding to transferrins

Fe(III), Cu(II), Co(III), and Mn(III) complexes of ovo- and human serum transferrins show resonance enhanced Raman bands near 1600, 1500, 1270, and 1170 cm-1 upon excitation with laser frequencies which fall within the visible absorption bands of those metalloproteins. Comparison of the visible absorption and resonance Raman spectra of the Cu(II)-transferrin complexes with those for the Cu(II) model compound, bis(2,4,6-trichlorophenolato)diimidazolecopper(II) monohydrate, indicates that the resonance Raman bands are due to enhancement of phenolic vibrational modes. For the model (Cu(II) compound, a normal coordinate analysis was used to aid our assignment of the observed resonance bands at 1562, 1463, 1311, and 1122 cm-1 to A1 vibrational modes of the 2,4,6-trichlorophenolato moiety. These assignments are consistent with those made for Cu(II)-transferrins. The latter assignments were based upon calculated A1 frequencies for p-methylphenol (Cummings, D.L., and Wood, J.L. (1974), J. Mol. Struct. 20, 1). The wavelength shifts in the resonance bands for the model compound from those for Cu(II)-transferrins are due to the influence of the chloro substituents on the planar vibrations of phenol. These results clearly identify tyrosine as a ligand in copper binding to transferrins.     

Properties of electrolyte solutions relevant to high concentration chloride leaching. III. Solubility of pertinent solids and iron(III)/iron(II) redox potential measured in concentrated magnesium chloride solutions

Results of solubility measurements of nickel chloride, manganese chloride, iron(II) chloride, hematite and akaganeite in aqueous solutions of MgCl₂ (0.5–3.5 mol L^− 1) at temperatures of 60 and 90 °C are reported. Solubilities of metal(II) chlorides decrease almost linearly with MgCl₂ concentration due to the common ion effect. Nickel chloride and iron(II) chloride solubilities are very similar, while manganese chloride is about 30% more soluble.Hematite is more stable (i.e. less soluble) than akaganeite under all conditions investigated in this study, while ferrihydrite is considerably less stable. In other words, there is no change in the relative stabilities of these phases effected by the presence of high magnesium chloride concentrations. The solubility of all of these phases decreases with temperature and, for each temperature, the solubility constants increase linearly with the MgCl₂ concentration. The present results allow the prediction of the iron concentration as a function of the H⁺ and MgCl₂ molality at equilibrium with hematite or akaganeite.The Fe(III)/Fe(II) redox behaviour has been characterized in concentrated aqueous solutions of MgCl₂ (1.5–3.5 mol L^− 1) at a temperature of 25 °C. Standard redox potentials are ca. 100 mV lower than at infinite dilution and change linearly by only 13 mV in the range 2–4 mol L^− 1 MgCl₂.     

The effect of Fe(II) and Fe(III) on the efficiency of copper electrowinning from dilute acid Cu(II) sulphate solutions with the chemelec cell. Part II. The efficiency of copper electrowinning from dilute liquors

This programme of work forms part of a study into the application of the Chemelec cell for direct electrowinning of copper from dilute leach liquors. A typical liquor is defined as containing less than 2 g l^?1 Cu(II), with an equivalent or greater concentration of iron as Fe(II) and Fe(III). Electrowinning experiments using simulated liquors showed that the current efficiency decreased in proportion to the increase in the Fe(III) concentration. A current efficiency of less than 30% was obtained during the electrowinning of a copper solution (1–2 g l^?1 Cu) with an Fe(III) concentration of 0.5–3.0 g l^?1. Electrowinning from leach liquors (1–2 g l^?1 Cu, 1–2 g l^?1 Fe(III) and 1–5 g l^?1 Fe(II)) showed improved efficiencies. A current efficiency of 47% was obtained for copper removal to less than 50 mg l^?1, with an electrolytic energy cost of about £198 per tonne at 3.3p per kWh. The impurity content of the copper was low, the most significant impurity being lead at 0.012%. This work has shown that the Chemelec cell can achieve reasonable efficiencies for direct electrowinning from dilute leach liquors. Further work is required in order to confirm the efficiency of electrowinning and to assess the operating and capital costs of a commercial cell.     

The effect of Fe(II) and Fe(III) on the efficiency of copper electrowinning from dilute acid Cu(II) sulphate solutions with the chemelec cell: Part I. Cathodic and anodic polarisation studies

The results are presented from an investigation into the effect of the Fe(III) and Fe(II) concentration on the current efficiency for copper electrowinning using the Chemelec cell. The results are from polarisation studies of acidic sulphate electrolytes containing Cu(II), Fe(II) and Fe(III). The mass transfer coefficients for these cationic species are determined, and the effect of using different anode materials on the overvoltage for the oxidation of Fe(II) and oxygen evolution is also examined.     

Effects of photoinhibition on the QA-Fe2+ complex of photosystem II studied by EPR and M?ssbauer spectroscopy

Effects of photoinhibition on the iron-quinone electron acceptor complex of oxygen-evolving photosystem II have been studied using low-temperature EPR and M?ssbauer spectroscopy. Photoinhibition of spinach photosystem II membrane particles at 4 degrees C decreases the EPR signal arising from the interaction of QA- with Fe2+ to 30% in 90 min under our conditions. The free radical EPR signal from QA- induced by cyanide treatment of the iron [Sanakis, Y., et al. (1994) Biochemistry 33, 9922-9928] declines with the same kinetics as the QA-Fe2+ EPR signal. In contrast, Fe2+ is present in about 70% of the centers after 90 min of photoinhibition, as shown by its EPR-detected interaction with NO and by its M?ssbauer absorption. Complete oxidation of this Fe2+ population to Fe3+ by ferricyanide is possible only in the presence of glycolate, which lowers the redox potential of the Fe3+/Fe2+ couple. In a fraction of PSII centers, which reach 30% after 90 min of photoinhibition, the iron cannot be detected. It is concluded that photoinhibition of oxygen-evolving photosystem II affects both QA and Fe2+. However, the photoinhibitory impairment of the QA redox functioning precedes the modification of the non-heme iron. In a considerable portion of the photoinhibited centers, which do not have functional QA, the non-heme iron is still present and redox active, but its redox potential is increased relative to that in the normal centers. This is probably due to a minor modification of the bicarbonate ligation site.(ABSTRACT TRUNCATED AT 250 WORDS)     

Interaction of Propagating Radicals with Copper(I) and Copper(II) Species

The effects of copper(I) and copper(II) metal centers on the atom transfer radical polymerization (ATRP) of styrene and methyl acrylate were investigated. The free-radical polymerizations were initiated by AIBN in the presence of copper(I) and copper(II) complexes. For methyl acrylate, the rate of the polymerization was reduced in the presence of CuIBr/dNbpy and CuIOTf/dTbpy but was unaffected by the presence of CuII(OTf)2/dTbpy. For styrene, under conditions which yield relatively low molecular weight polymer (16 000), no effect was observed in the presence of CuII(OTf)2/dNbpy; however, under conditions which yield high molecular weight polystyrene (50 000-100 000), the polymerization was limited in the molecular weight attainable and stopped at partial conversion. No effect was observed for the free-radical polymerization of styrene in the presence of copper(I) complexes. These results indicate that control in ATRP does not originate in interactions of growing radicals with copper complexes but in the reversible halogen atom transfer.     

Recovery of copper(II) from aqueous solutions by means of supported liquid membranes

Relatively stable liquid membranes were easily and reproducibly prepared by impregnating commercial porous polytetrafluoroethylene or polypropylene membranes with solutions of commercial chelating extractants. When such membranes were placed between aqueous solutions of copper(II) sulfate at pH 1–4 and of sulfuric acid at pH 0.5, transfer of copper(II) ion from the less acidic to the more acidic solution took place simultaneously with an equivalent proton transfer in the opposite direction. The copper(II) ion moved selectively with respect to iron(III) ion and against a twenty-fold concentration gradient. The mechanistic and practical implications of these results are discussed.     

Solubilities,densities and electrical conductivities of aqueous copper(I) and copper(II) chlorides in solutions containing other chlorides such as iron,zinc, sodium and hydrogen chlorides

With a view to industrial applications, solubilities, densities and electrical conductivities of aqueous copper(I) and copper(II) chlorides were measured in solutions containing other chlorides like those of iron, zinc, sodium and hydrogen.In these solutions copper(I) chloride shows a behaviour similar to that observed in pure CuClNaClHCl solutions. Increasing the FeCl₂ concentration decreases the solubility of CuCl less than increasing the ZnCl₂ concentrations does. Moreover, increasing total Cl^? concentration increases CuCl solubility if Cl^? is added as FeCl₂ and decreases CuCl solubility if Cl^? is added as ZnCl₂. NaCl solubility remains unchanged by FeCl₂ additions and is increased by ZnCl₂ additions.Copper(II) chloride also shows in these solutions a behaviour similar to that observed in pure CuCl₂NaClHCl solutions. FeCl₃ additions decrease CuCl₂ · 2 H₂O solubility in a more drastic manner than ZnCl₂ additions. However, NaCl additions have a much higher effect.These results can be qualitatively interpreted when taking into account the relative Cl^? donor or acceptor character of the constituent chloride salts of the solution.FeCl₂ additions slightly decrease the electrical conductivity of Cu(I) solutions. However, this can easily be compensated by a slight increase in temperature or in acidity.     

The coprecipitation behavior of Co(II) and Ni(II) with Fe(III), Cr(III) and Al(III) from aqueous ammoniacal solutions

The coprecipitation of cobalt(II) and nickel(II) with iron(III), chromium(III) and aluminum(III) from ammoniacal solutions has been investigated. The coprecipitation behavior was found to be very sensitive to the solution pH and total ammonia concentration. Co(II) and Ni(II) can be precipitated from low ammonia concentration solutions but are readily redissolved at higher ammonia concentrations. The coprecipitate of divalent and trivalent species was found to contain very large amounts of the divalent metals (up to a mole ratio M M(II)/M(III) of 2.5) when aluminum was the trivalent species, whereas with iron(III) or chromium(III), the ratio was only 0.5.     

Cuprous hydrometallurgy Party VI. Activation of chalcopyrite by reduction with copper and solutions of copper(I) salts

Chalcopyrite is reduced by solutions of copper(I) sulfate and copper(I) chloride to chalcocite (Cu₂S) and bornite (Cu₅FeS₄) whilst the iron reports to the solution. Factors which affect the rate and efficiency of reduction are examined. The reaction is rapid on fresh surfaces of chalcopyrite but slows markedly as a film of chalcocite or bornite forms. The reduction in the presence of copper metal goes to completion and gives a material which is more readily leached by oxidising agents than is chalcopyrite. Thus 99% of the copper in the reduced chalcopyrite is leached when copper(II) sulfate in aqueous acetonitrile is the oxidising agent, whereas only 30% of the copper is leached from pure chalcopyrite under similar conditions. Concentrated solutions of copper(I) salts are less effective in reducing CuFeS₂ in a heterogeneous solid-liquid reaction than is copper metal in a “galvanic” solid-solid reaction. Solutions of copper(II) sulfate plus concentrated copper(I) sulfate in dilute acetonitrile (4 M) containing copper sheets are an effective reductant for chalcopyrite.     

Extraction of nickel,cobalt and other metals [Cu,Zn, Fe(III)] with a commercial β-diketone extractant

The extraction of nickel, cobalt, copper and zinc from ammoniacal solutions of ammonium carbonate or ammonium sulphate by solutions of Hostarex DK-16 in kerosene has been investigated as a function of phase contact time, aqueous-phase pH and organicphase reagent concentration. Besides copper, Hostarex DK-16 also partially extracts iron (III) from moderately acidic solutions whereas nickel, cobalt(II), copper and zinc are extracted from neutral or ammoniacal ammonium sulphate and ammonium carbonate solutions. Extraction decreases in the following order of metals: Cu > Co > Ni > Zn. Cobalt(III) is not extracted, but the complex of cobalt(II) with Hostarex DK-16 is slowly oxidized to a cobalt(III) complex which cannot be stripped even when 10 N sulphuric acid is used. Absorption spectra for cobalt complexes with Hostarex DK-16 (purified by preparative thin-layer chromatography) in benzene also suggest oxidation of cobalt(II) to cobalt(III) in the organic phase. Nickel, cobalt(II), zinc and copper can be stripped easily from organic solution with dilute solutions of sulphuric acid. Hostarex DK-16 extracts iron very slowly, nickel moderately rapidly and copper, cobalt(II) and zinc rapidly. Slope analysis and extraction isotherms suggest that the complexes CuR₂, NiR₂ ·HR and CoR₂·HR are present in the organic phase. Nickel can easily be separated from cobalt by extraction with Hosterex DK-16 after oxidation of cobalt in aqueous ammoniacal solution by hydrogen peroxide; however, LIX 64N seems to be a more promising extractant owing to the higher extraction of nickel under analogous conditions and the poorer extraction of zinc in comparison with Hostarex DK-16.     

Conflict and collaboration in middle-aged and older couples: II. Cardiovascular reactivity during marital interaction.

Marital strain confers risk of cardiovascular disease (CVD), perhaps though cardiovascular reactivity (CVR) to stressful marital interactions. CVR to marital stressors may differ between middle-age and older adults, and types of marital interactions that evoke CVR may also differ across these age groups, as relationship contexts and stressors differ with age. The authors examined cardiovascular responses to a marital conflict discussion and collaborative problem solving in 300 middle-aged and older married couples. Marital conflict evoked greater increases in blood pressure, cardiac output, and cardiac sympathetic activation than did collaboration. Older couples displayed smaller heart rate responses to conflict than did middle-aged couples but larger blood pressure responses to collaboration—especially in older men. These effects were maintained during a posttask recovery period. Women did not display greater CVR than men on any measure or in either interaction context, though they did display greater parasympathetic withdrawal. CVR to marital conflict could contribute to the association of marital strain with CVD for middle-aged and older men and women, but other age-related marital contexts (e.g., collaboration among older couples) may also contribute to this mechanism. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Competitive solvation and complexation of Cu(I), Cu(II), Pb(II), Zn(II), and Ag(I) in aqueous ethanol,acetonitrile, and dimethylsulfoxide solutions containing chloride ion with applications to hydrometallurgy

The changes in reduction potential and single ion activity of Cu(I), Cu(II), Pb(II), Zn(II), and Ag(I) have been measured in a range of aqueous ethanol (EtOH), acetonitrile (AN), and dimethylsulfoxide (DMSO) compositions containing excess chloride ion. The results are compared with changes in such solutions in the absence of chloride ion and with the changes in strong brines and rationalized in terms of the various competitive ion-solvent and ion-chloride interactions. Organic solvents are shown to generally enhance chloride ion activity and promote complex ion formation. But AN is a stronger ligand for Cu(I) and DMSO is a stronger ligand for Cu(II) and Zn(II) than is Cl⁻ or the other solvents. The decrease in metal ion activity in mixed aqueous solvents containing Cl⁻ is greater than that in concentrated aqueous chloride salt solutions, according to the strength of the chloro- or solvo-complex. These fundamental changes lead to applications in the extraction of metal ion complexes and promote the dissolution of AgCl, PbCl₂, and CuCl in aqueous DMSO containing Cl⁻. Formerly Doctoral Student, Murdoch University     

Electrochemical modeling and study of copper deposition from concentrated ammoniacal sulfate solutions

A fundamental investigation of the electrolytic deposition of copper from concentrated aqueous ammoniacal solutions has been carried out based on the thermodynamic analysis of the system Cu–NH₃–H₂O. The speciation of copper vs. pH and redox potential was modeled in high ionic strength solutions, in which the activity coefficients of the system species were estimated according to the Modified Bromley's Methodology. The electrochemical behavior of the redox system Cu(0)/Cu(I)/Cu(II) in concentrated aqueous ammoniacal solutions was studied at pH = 9.5 and the cathodic reactions in these solutions were determined. It was found that metallic copper was formed under strongly reductive redox conditions, while under mildly reductive to mildly oxidative conditions the cuprous di-ammine complex species dominate. Under highly oxidative conditions the cupric tetra-ammine complex species predominated. According to the theory and results, the cathodic deposition of copper from concentrated aqueous ammoniacal solutions proceeds in a two-step reduction mechanism. The cupric ammine species are first reduced to cuprous di-ammine, which in turn is reduced to metallic copper. The electrochemical experiments revealed that copper deposition over time follows a sigmoid-type curve, verifying the two-step mechanism. The main feature of these sigmoid curves was the presence of an induction period with negligible copper deposition, followed by an acceleration period where the copper deposition rate gradually increased. By increasing the applied cell voltage, the induction period was significantly reduced or disappeared.     

Marital interaction in alcoholic and nonalcoholic couples: Alcoholic subtype variations and wives' alcoholism status.

The authors examined problem-solving marital interactions of alcoholic and nonalcoholic couples (N = 132). Four alcoholic groups (husband alcoholic with antisocial personality disorder or not, paired with alcoholic or nonalcoholic wives) were compared with each other and with a both-spouses-nonalcoholic group. Consistent with the alcoholic subtypes hypothesis, couples with an antisocial alcoholic husband had higher levels of hostile behavior regardless of wives' alcoholism status. In contrast, rates of positive behaviors and the ratio of positive to negative behaviors were greatest among couples in which either both or neither of the spouses had alcoholic diagnoses and were lowest among alcoholic husbands with nonalcoholic wives. Discussion focuses on possible mechanisms linking antisocial alcoholism and discrepant alcoholic diagnoses to poorer marital outcomes. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Metal-catalyzed oxidation and mutagenesis studies on the iron(II) binding site of 1-aminocyclopropane-1-carboxylate oxidase

The final step in the biosynthesis of the plant signaling molecule ethylene is catalyzed by 1-aminocyclopropane-1-carboxylate (ACC) oxidase, a member of the non-heme iron(II) dependent family of oxygenases and oxidases, which has a requirement for ascorbate as a co-substrate and carbon dioxide as an activator. ACC oxidase (tomato) has a particularly short half-life under catalytic conditions undergoing metal-catalyzed oxidative (MCO) fragmentation. Sequence comparisons of ACC oxidases with isopenicillin N synthase (IPNS) and members of the 2-oxoglutarate Fe(II) dependent dioxygenases show an aspartate and two of six ACC oxidase conserved histidine residues are completely conserved throughout this subfamily of Fe(II) dependent oxygenases/oxidases. Previous mutagenesis, spectroscopic, and crystallographic studies on IPNS indicate that the two completely conserved histidine and aspartate residues act as Fe(II) ligands. To investigate the role of the conserved aspartate and histidine residues in ACC oxidase (tomato fruit), they were substituted via site-directed mutagenesis. Modified ACC oxidases produced were H39Q, H56Q, H94Q, H177Q, H177D, H177E, D179E, D179N, H177D&D179E, H211Q, H234Q, H234D, and H234E. Among those histidine mutants replaced by glutamine, H39Q, H56Q, H94Q, and H211Q were catalytically active, indicating these histidines are not essential for catalysis. Mutant enzymes H177D, H177Q, D179N, H177D&D179E, H234Q, H234D, and H234E were catalytically inactive consistent with the assignment of H177, D179, and H234 as iron ligands. Replacement of H177 with glutamate or D179 with glutamate resulted in modified ACC oxidases which still effected the conversion of ACC to ethylene, albeit at a very low level of activity, which was stimulated by bicarbonate. The H177D (inactive), H177E (low activity), D179E (low activity), and H234Q (inactive) modified ACC oxidases all underwent MCO fragmentation, indicating that they can bind iron, dioxygen, ACC, and ascorbate. The results suggest that MCO cleavage results from active site-mediated reactions and imply that, while H177, D179, and H234 are all involved in metal ligation during catalysis, ligation to H234 is not required for fragmentation. It is possible that MCO fragmentation results from reaction of incorrectly folded or "primed" ACC oxidase.     

厌氧环境下邻菲罗啉分光光度法测定纤维水镁石中Fe(II)与Fe(III)

纤维水镁石中同时存在有不同价态的Fe即Fe(II)与Fe(III),这对开展水镁石相关研究具有重要意义。而不同价态Fe含量测定过程中存在Fe(II)在空气中易氧化成Fe(III)而导致测定不准确的问题。针对该问题,实验采用厌氧培养箱作为操作环境,对纤维水镁石的处理均在厌氧条件下进行。利用邻菲罗啉与经盐酸羟胺还原产生的总Fe(II)显色反应原理测定纤维水镁石中的总Fe,之后测定过程中不加盐酸羟胺与邻菲罗啉反应测定溶液中的Fe(II),利用总Fe与Fe(II)的差值测定Fe(III),从而实现了邻菲罗啉分光光度法对纤维水镁石中Fe(II)与Fe(III) 的准确测定。试验发现在波长为510nm,显色体系pH值为2~5,显色时间为10min条件下,Fe(II)质量浓度在0.02~5.0mg/L范围内与其吸光度符合比尔定律,校准曲线的相关系数为0.9999。采用实验方法对两个纤维水镁石样品分别进行6次平行测定,结果显示总Fe、Fe(II)及Fe(III)的相对标准偏差(RSD,n=6)均小于1%。采用电感耦合等离子体原子发射光谱法(ICP-AES)进行全Fe测定结果对照,结果表明两种方法的测定结果保持一致。     

Spectral and kinetic properties of the Fet3 protein from Saccharomyces cerevisiae, a multinuclear copper ferroxidase enzyme

High affinity iron uptake in Saccharomyces cerevisiae requires Fet3p. Fet3p is proposed to facilitate iron uptake by catalyzing the oxidation of Fe(II) to Fe(III) by O2; in this model, Fe(III) is the substrate for the iron permease, encoded by FTR1. Here, a recombinant Fet3p has been produced in yeast that, lacking the C-terminal membrane-spanning domain, is secreted directly into the growth medium. Solutions of this Fet3p at >1 mg/ml have the characteristic blue color of a type 1 Cu(II)-containing protein, consistent with the sequence homology that placed this protein in the class of multinuclear copper oxidases that includes ceruloplasmin. Fet3p has an intense absorption at 607 nm (epsilon = 5500 M-1 cm-1) due to this type 1 Cu(II) and a shoulder in the near UV at 330 nm (epsilon = 5000 M-1 cm-1) characteristic of a type 3 binuclear Cu(II) cluster. The EPR spectrum of this Fet3p showed the presence of one type 1 Cu(II) and one type 2 Cu(II) (A parallel = 91 and 190 x 10(-4) cm-1, respectively). Copper analysis showed this protein to have 3.85 g atom copper/mol, consistent with the presence of one each of the three types of Cu(II) sites found in multinuclear copper oxidases. N-terminal analysis demonstrated that cleavage of a signal peptide occurred after Ala-21 in the primary translation product. Mass spectral and carbohydrate analysis of the protein following Endo H treatment indicated that the preparation was still 15% (w/w) carbohydrate, probably O-linked. Kinetic analysis of the in vitro ferroxidase reaction catalyzed by this soluble Fet3p yielded precise kinetic constants. The Km values for Fe(II) and O2 were 4.8 and 1.3 microM, respectively, while kcat values for Fe(II) and O2 turnover were 9.5 and 2.3 min-1, consistent with an Fe(II):O2 reaction stoichiometry of 4:1.