Heterologous Expression and Metal-Binding Characterization of a Type 1 Metallothionein Isoform (OsMTI-1b) from Rice (Oryza sativa)

Metallothioneins (MTs) are ubiquitous, low molecular mass and cysteine-rich proteins that play important roles in maintaining intracellular metal homeostasis, eliminating metal toxification and protecting the cells against oxidative damages. MTs are able to bind metal ions through the thiol groups of their cysteine residues. Plants have several MT isoforms which are classified into four types based on the arrangement of cysteine residues. In the present study, a rice (Oryza sativa) gene encoding type 1 MT isoform, OsMTI-1b, was inserted in vector pET41a and overexpressed in Escherichia coli as carboxy-terminal extensions of glutathione-S-transferase (GST). The recombinant protein GST-OsMTI-1b was purified using affinity chromatography and its ability to bind with Ni²⁺, Cd²⁺, Zn²⁺ and Cu²⁺ ions was analyzed. The results demonstrated that this isoform has ability to bind Ni²⁺, Cd²⁺ and Zn²⁺ ions in vitro, whereas it has no substantial ability to bind Cu²⁺ ions. From competitive reaction with 5,5′-dithiobis(2-nitrobenzoic acid), DTNB, the affinity of metal ions for recombinant form of GST-OsMTI-1b was as follows: Ni²⁺/Cd²⁺ > Zn²⁺ > Cu²⁺     

Isolation,Growth, Ultrastructure,and Metal Tolerance of the Green Alga,Chlamydomonas acidophila (Chlorophyta)

An acidophilic volvocine flagellate, Chlamydomonas acidophila (Volvocales) that was isolated from an acid lake, Katanuma, in Miyagi prefecture, Japan was studied for growth, ultrastructural characterization, and metal tolerance.

Chlamydomonas acidophila is obligately photoautotrophic, and did not grow in the cultures containing acetate or citrate even in the light. The optimum pH for growth was 3.5-4.5. To characterize metal tolerance, the toxic effects of Cd, Co, Cu, and Zn on this alga were also studied. Effective metal concentrations, which limited the growth by 50%, EC₅₀ were measured, after 72h of static exposure. EC₅₀s were 14.4 μM Cd²⁺, 81.3 μM Co²⁺, 141μM Cu²⁺, and 1.16 mM Zn²⁺ for 72 h of exposure. Thus, this alga had stronger tolerance to these metals than other species in the genus Chlamydomonas.     

Heavy metal ions affect the activity of DNA glycosylases of the Fpg family

Prokaryotic enzymes formamidopyrimidine-DNA glycosylase (Fpg) and endonuclease VIII (Nei) and their eukaryotic homologs NEIL1, NEIL2, and NEIL3 define the Fpg family of DNA glycosylases, which initiate the process of repair of oxidized DNA bases. The repair of oxidative DNA lesions is known to be impaired in vivo in the presence of ions of some heavy metals. We have studied the effect of salts of several alkaline earth and transition metals on the activity of Fpg-family DNA glycosylases in the reaction of excision of 5,6-dihydrouracil, a typical DNA oxidation product. The reaction catalyzed by NEIL1 was characterized by values K _m = 150 nM and k _cat = 1.2 min⁻¹, which were in the range of these constants for excision of other damaged bases by this enzyme. NEIL1 was inhibited by Al³⁺, Ni²⁺, Co²⁺, Cd²⁺, Cu²⁺, Zn²⁺, and Fe²⁺ in Tris-HCl buffer and by Cd²⁺, Zn²⁺, Cu²⁺, and Fe²⁺ in potassium phosphate buffer. Fpg and Nei, the prokaryotic homologs of NEIL1, were inhibited by the same metal ions as NEIL1. The values of I₅₀ for NEIL1 inhibition were 7 μM for Cd²⁺, 16 μM for Zn²⁺, and 400 μM for Cu²⁺. The inhibition of NEIL1 by Cd²⁺, Zn²⁺, and Cu²⁺ was at least partly due to the formation of metal-DNA complexes. In the case of Cd²⁺ and Cu²⁺, which preferentially bind to DNA bases rather than phosphates, the presence of metal ions caused the enzyme to lose the ability for preferential binding to damaged DNA. Therefore, the inhibition of NEIL1 activity in removal of oxidative lesions by heavy metal ions may be a reason for their comutagenicity under oxidative stress.     

Prion metal interaction: Is prion pathogenesis a cause or a consequence of metal imbalance?

Functional role of cellular prion protein (PrPc) has been hypothesized to be in metal homeostasis and providing cells with a superoxide dismutase (SOD)-like activity to escape damage by reactive oxygen species (ROS). PrPc interacts with a range of divalent metal ions and undergoes Cu²⁺ as well as Zn²⁺-associated endocytosis, thereby maintaining homeostasis of these and other metal ions. Conformational change to a β-sheet rich, protease resistant entity, reminiscent of the disease-associated scrapie form called PrPsc, has been found to be induced by interaction of PrPc with metal ions like Cu²⁺, Zn²⁺, Mn²⁺ and Fe²⁺. This review compiles data from various experimental studies of the interaction of metals with PrPc. The effect of metal ions on the expression and conformation of the prion protein is described in detail with emphasis on their possible physiological and pathogenic role. Further, a hypothesis is presented where attainment of altered conformation by metal-bound PrPc has been viewed as a deleterious consequence of efforts made by cells to maintain metal homeostasis. Thus, PrPc presumably sacrifices itself by converting into PrPsc form in an attempt to protect cells from the toxicity of metal imbalance. Finally, possible reasons for contradictions reported in the literature on the subject are explored and experimental approaches to resolve the same are suggested.     

Nickel in plants: I. Uptake kinetics using intact soybean seedlings

The absorption of Ni²⁺ by 21-day-old soybean plants (Glycine max cv. Williams) was investigated with respect to its concentration dependence, transport kinetics, and interactions with various nutrient cations. Nickel absorption, measured as a function of concentration (0.02 to 100 μm), demonstrated the presence of multiple absorption isotherms. Each of the three isotherms conforms to Michaelis-Menten kinetics; kinetic constants are reported for uptake by the intact plant and for transfer from root to shoot tissues. The absorption of Ni²⁺ by the intact plant and its transfer from root to shoot were inhibited by the presence of Cu²⁺, Zn²⁺, Fe²⁺, and Co²⁺. Competition kinetic studies showed Cu²⁺ and Zn²⁺ to inhibit Ni²⁺ absorption competitively, suggesting that Ni²⁺, Cu²⁺, and Zn²⁺ are absorbed using the same carrier site. Calculated K_m and K_i constants for Ni²⁺ in the presence and absence of Cu²⁺ were 6.1 and 9.2 μm, respectively, whereas K_m and K_i constants were calculated to be 6.7 and 24.4 μm, respectively, for Ni²⁺ in the presence and absence of Zn²⁺. The mechanism of inhibition of Ni²⁺ in the presence of Fe²⁺ and Co²⁺ was not resolved by classical kinetic relationships.     

Substrate Profile and Metal-ion Selectivity of Human Divalent Metal-ion Transporter-1

Divalent metal-ion transporter-1 (DMT1) is a H⁺-coupled metal-ion transporter that plays essential roles in iron homeostasis. DMT1 exhibits reactivity (based on evoked currents) with a broad range of metal ions; however, direct measurement of transport is lacking for many of its potential substrates. We performed a comprehensive substrate-profile analysis for human DMT1 expressed in RNA-injected Xenopus oocytes by using radiotracer assays and the continuous measurement of transport by fluorescence with the metal-sensitive PhenGreen SK fluorophore. We provide validation for the use of PhenGreen SK fluorescence quenching as a reporter of cellular metal-ion uptake. We determined metal-ion selectivity under fixed conditions using the voltage clamp. Radiotracer and continuous measurement of transport by fluorescence assays revealed that DMT1 mediates the transport of several metal ions that were ranked in selectivity by using the ratio I_max/K_0.5 (determined from evoked currents at −70 mV): Cd²⁺ > Fe²⁺ > Co²⁺, Mn²⁺ ≫ Zn²⁺, Ni²⁺, VO²⁺. DMT1 expression did not stimulate the transport of Cr²⁺, Cr³⁺, Cu⁺, Cu²⁺, Fe³⁺, Ga³⁺, Hg²⁺, or VO⁺. ⁵⁵Fe²⁺ transport was competitively inhibited by Co²⁺ and Mn²⁺. Zn²⁺ only weakly inhibited ⁵⁵Fe²⁺ transport. Our data reveal that DMT1 selects Fe²⁺ over its other physiological substrates and provides a basis for predicting the contribution of DMT1 to intestinal, nasal, and pulmonary absorption of metal ions and their cellular uptake in other tissues. Whereas DMT1 is a likely route of entry for the toxic heavy metal cadmium, and may serve the metabolism of cobalt, manganese, and vanadium, we predict that DMT1 should contribute little if at all to the absorption or uptake of zinc. The conclusion in previous reports that copper is a substrate of DMT1 is not supported.     

Heavy metal uptake capacities by the common freshwater green alga Cladophora fracta

Macroalgae have received much attention for heavy metal removal in treatment of domestic wastewater. In this report, the uptake capacity of a common freshwater green alga, Cladophora fracta, for heavy metal ions (copper, zinc, cadmium, and mercury) was evaluated. The equilibrium adsorption capacities were 2.388?mg Cu²⁺, 1.623?mg Zn²⁺, 0.240?mg Cd²⁺, and 0.228?mg Hg²⁺ per gram of living algae at 18°C and pH?5.0. The removal efficiency for Cu²⁺, Zn²⁺, Cd²⁺, and Hg²⁺ were 99, 85, 97, and 98%, respectively. Greater removal efficiency was achieved when the concentrations of metal ions were at very low level. The results indicated that living algae are suitable for removal and recovery of heavy metal ions from aqueous solutions and can be a potential tool to treat industrial wastewater.     

Multi-metal biosorption and bioaccumulation by Exiguobacterium sp. ZM-2

The present work deals with the biosorption performance of dried and non-growing biomasses of Exiguobacterium sp. ZM-2, isolated from soil contaminated with tannery effluents, for the removal of Cd²⁺, Ni²⁺, Cu²⁺, and Zn²⁺ from aqueous solution. The metal concentrations studied were 25 mg/l, 50 mg/l, 100 mg/l, 150 mg/l and 200 mg/l. The effect of solution pH and contact time was also studied. The biosorption capacity was significantly altered by pH of the solution. The removal of metal ions was conspicuously rapid; most of the total sorption occurred within 30 min. The sorption data have been analyzed and fitted to the Langmuir and Freundlich isotherm models. The highest Q_max value was found for the biosorption of Cd²⁺ at 43.5 mg/g in the presence of the non-growing biomass. Recovery of metals (Cd²⁺, Zn²⁺, Cu²⁺ and Ni²⁺) was found to be better when dried biomass was used in comparison to non-growing biomass. Metal removal through bioaccumulation was determined by growing the bacterial strain in nutrient broth amended with different concentrations of metal ions. This multi-metal resistant isolate could be employed for the removal of heavy metals from spent industrial effluents before discharging them into the environment.     

Reversible immobilization of invertase on Cu-chelated polyvinylimidazole-grafted iron oxide nanoparticles

Polyvinylimidazole (PVI)-grafted iron oxide nanoparticles (PVIgMNP) were prepared by grafting of telomere of PVI on the iron oxide nanoparticles. Different metal ions (Cu²⁺, Zn²⁺, Cr²⁺, Ni²⁺) ions were chelated on polyvinylimidazole-grafted iron oxide nanoparticles, and then the metal-chelated magnetic particles were used in the adsorption of invertase. The maximum invertase immobilization capacity of the PVIgMNP–Cu²⁺ beads was observed to be 142.856 mg/g (invertase/PVIgMNP) at pH 5.0. The values of the maximum reaction rate (V _max) and Michaelis–Menten constant (Km) were determined for the free and immobilized enzymes. The enzyme adsorption–desorption studies, pH effect on the adsorption efficiency, affinity of different metal ions, the kinetic parameters and storage stability of free and immobilized enzymes were evaluated.     

Effect of metal ions (Li+, Na+, K+, Mg2+, Ca2+, Ni2+, Cu2+ and Zn2+) and water coordination on the structure and properties of l-histidine and zwitterionic l-histidine

Interactions between metal ions and amino acids are common both in solution and in the gas phase. The effect of metal ions and water on the structure of l-histidine is examined. The effect of metal ions (Li⁺, Na⁺, K⁺, Mg²⁺, Ca²⁺, Ni²⁺, Cu²⁺ and Zn²⁺) and water on structures of His·M(H₂O)m, m = 0.1 complexes have been determined theoretically employing density functional theories using extended basis sets. Of the five stable complexes investigated the relative stability of the gas-phase complexes computed with DFT methods (with one exception of K⁺ systems) suggest metallic complexes of the neutral l-histidine to be the most stable species. The calculations of monohydrated systems show that even one water molecule has a profound effect on the relative stability of individual complexes. Proton dissociation enthalpies and Gibbs energies of l-histidine in the presence of the metal cations Li⁺, Na⁺, K⁺, Mg²⁺, Ca²⁺, Ni²⁺, Cu²⁺ and Zn²⁺ were also computed. Its gas-phase acidity considerably increases upon chelation. Of the Lewis acids investigated, the strongest affinity to l-histidine is exhibited by the Cu²⁺ cation. The computed Gibbs energies ΔG are negative, span a rather broad energy interval (from ?130 to ?1,300 kJ/mol), and upon hydration are appreciably lowered.     

Heterogeneous behavior of metalloproteins toward metal ion binding and selectivity: insights from molecular dynamics studies

About one-third of the existing proteins require metal ions as cofactors for their catalytic activities and structural complexities. While many of them bind only to a specific metal, others bind to multiple (different) metal ions. However, the exact mechanism of their metal preference has not been deduced to clarity. In this study, we used molecular dynamics (MD) simulations to investigate whether a cognate metal (bound to the structure) can be replaced with other similar metal ions. We have chosen seven different proteins (phospholipase A₂, sucrose phosphatase, pyrazinamidase, cysteine dioxygenase (CDO), plastocyanin, monoclonal anti-CD4 antibody Q425, and synaptotagmin 1 C₂B domain) bound to seven different divalent metal ions (Ca²⁺, Mg²⁺, Zn²⁺, Fe²⁺, Cu²⁺, Ba²⁺, and Sr²⁺, respectively). In total, 49 MD simulations each of 50 ns were performed and each trajectory was analyzed independently. Results demonstrate that in some cases, cognate metal ions can be exchanged with similar metal ions. On the contrary, some proteins show binding affinity specifically to their cognate metal ions. Surprisingly, two proteins CDO and plastocyanin which are known to bind Fe²⁺ and Cu²⁺, respectively, do not exhibit binding affinity to any metal ion. Furthermore, the study reveals that in some cases, the active site topology remains rigid even without cognate metals, whereas, some require them for their active site stability. Thus, it will be interesting to experimentally verify the accuracy of these observations obtained computationally. Moreover, the study can help in designing novel active sites for proteins to sequester metal ions particularly of toxic nature.     

A spectrophotometric method for the determination of zinc, copper, and cobalt ions in metalloproteins using Zincon

Zincon (2-carboxy-2′-hydroxy-5′-sulfoformazylbenzene) has long been known as an excellent colorimetric reagent for the detection of zinc and copper ions in aqueous solution. To extend the chelator’s versatility to the quantification of metal ions in metalloproteins, the spectral properties of Zincon and its complexes with Zn²⁺, Cu²⁺, and Co²⁺ were investigated in the presence of guanidine hydrochloride and urea, two common denaturants used to labilize metal ions in proteins. These studies revealed the detection of metals to be generally more sensitive with urea. In addition, pH profiles recorded for these metals indicated the optimal pH for complex formation and stability to be 9.0. As a consequence, an optimized method that allows the facile determination of Zn²⁺, Cu²⁺, and Co²⁺ with detection limits in the high nanomolar range is presented. Furthermore, a simple two-step procedure for the quantification of both Zn²⁺ and Cu²⁺ within the same sample is described. Using the prototypical Cu²⁺/Zn²⁺-protein superoxide dismutase as an example, the effectiveness of this method of dual metal quantification in metalloproteins is demonstrated. Thus, the spectrophotometric determination of metal ions with Zincon can be exploited as a rapid and inexpensive means of assessing the metal contents of zinc-, copper-, cobalt-, and zinc/copper-containing proteins.     

Binding of divalent cations with poly (S-carboxymethyl-l-cysteine) and their effects on the polypeptide conformation

The effects of divalent ions on fully charged $\text{poly}\text{(S-}\text{carboxymethyl-}\text{l}\text{-cysteine}$) have been examined using circular dichroism for eight species: CuCl₂, CdCl₂, ZnCl₂, NiCl₂, CoCl₂, BaCl₂, CaCl₂, and MgCl₂. Five of them are effective inducers of β-form in the order: Cu²⁺Cd²⁺Zn²⁺Ni²⁺Co²⁺, in media with no salt added. However, the other three ions (Ba²⁺, Ca²⁺ and Mg²⁺), are not effective. Precipitation occurs when metal chlorides reach the vicinity of the equivalent point except for Ca²⁺ and Mg²⁺. Precipitation of the random coil form is slow, while that of the β-form is rapid. Addition of NcCl reduces the solubility of the β-form considerably. The pH value varies linearly with the logarithm of metal chloride concentration C_M for Ba²⁺, Ca²⁺ and Mg²⁺ ions, while nonlinear dependence of pH on log C_M is found for Cu²⁺, Ni²⁺ and Co²⁺ ions.     

Topochemical factors in potentiation of contraction by heavy metal cations

In addition to the previously studied Zn²⁺, low concentrations (about 0.5 mM) of Be²⁺, Ba²⁺, Cd²⁺, Ni²⁺, Cu²⁺, Pt⁴⁺ and, outstandingly, 0.5 µM of UO₂ ²⁺, potentiate the twitch of frog sartorius and toe muscles by prolonging the active state of contraction. The degree of potentiation is a roughly S-shaped function of p(metal²⁺), suggesting that each metal binds to a ligand of the muscle fiber, representative apparent affinity constants being: UO₂ ²⁺, 5 x 10⁶; Zn²⁺, 2.8 x 10⁵; and Cd²⁺, 2 x 10⁴. UO₂ ²⁺ potentiation effects are rapidly reversed by PO₄, and Zn²⁺ and Cd²⁺ effects by EDTA, PO₄, and cysteine. The rapidity of these reversals by the nonpenetrating EDTA and PO₄, and the fact that heavy metal ions evidently potentiate by prolonging the action potential, indicate that the metal potentiators exert their primary action at readily accessible (i.e. plasma and T tubular) membrane sites. The relatively slow kinetics of development of potentiation, and the even slower reversal of it in pure Ringer''s solution, indicate that the metal ions are bound to connective tissue, as well as to muscle fibers. The binding effects at the readily accessible membrane sites evidently impairs delayed rectification and thus modifies the action potential and excitation-contraction coupling so as to cause potentiation. SH is excluded, and PO₄ and imidazole are possibilities, as the membrane ligand binding the potentiating metal ions.     

Metal Ions and Electrolytes Regulate the Dissociation of Heme from Human Hemopexin at Physiological pH

The stability of the hemopexin-heme (Hx-heme) complex to dissociation of the heme prosthetic group has been examined in bicarbonate buffers in the presence and absence of various divalent metal ions. In NH₄HCO₃ buffer (pH 7.4, 20 mm, 25 °C) containing Zn²⁺ (100 μm), 14% of the heme dissociates from this complex (4.5 μm) within 10 min, and 50% dissociates within 2 h. In the absence of metal ions, the rate of dissociation of this complex is far lower, is decreased further in KHCO₃ solution, and is minimal in NaHCO₃. In NH₄HCO₃ buffer, dissociation of the Hx-heme complex is accelerated by addition of divalent metals with decreasing efficiency in the order Zn²⁺ > Cu²⁺ ≫ Ni²⁺ > Co²⁺≫Mn²⁺. Addition of Ca²⁺ prior to addition of Zn²⁺ stabilizes the Hx-heme complex to dissociation of the heme group, and addition of Ca²⁺ after Zn²⁺-induced dissociation of the Hx-heme complex results in re-formation of the Hx-heme complex. These effects are greatly accelerated at 37 °C and diminished in other buffers. Overall, the solution conditions that promote formation of the Hx-heme complex are similar to those found in blood plasma, and conditions that promote release of heme are similar to those that the Hx-heme complex should encounter in endosomes following endocytosis of the complex formed with its hepatic receptor.     

Effect of heavy metal ions on growth and biochemical characteristics of photosynthesis of barley and maize seedlings

The effects of Cu²⁺, Zn²⁺, Cd²⁺ and Pb²⁺ on growth and the biochemical characteristics of photosynthesis were more expressed in barley (Hordeum vulgare L.) than in maize (Zea mays L.) seedlings. The barley and maize seedlings exhibited retardation in shoot and root growth after exposure of Cu²⁺, Cd²⁺ and Pb²⁺. The Zn²⁺ions practically did not influence these characteristics. The total protein content of barley and maize roots declined with an increase in heavy metal ion concentrations. The protein content of barley shoots was only slighly decreased with an increase in heavy metal ion concentrations, but the protein content in maize shoots was increased under the same conditions. The chlorophyll content was decreased in barley shoots and increased in maize. The ribulose-l,5-bisphosphate carboxylase (RuBPC, EC 4.1.1.39) and phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) activities were decreased drastically by Cu²⁺, Cd²⁺ and Pb²⁺ in thein vivo experiments. The tested heavy metal ions affect photosynthesis probably mainly by inhibition of these key carboxylating enzymes: this mechanism was studied in thein vitro experiments.     

Fe2+ binding on amyloid β‐peptide promotes aggregation

The metal ions Zn²⁺, Cu²⁺, and Fe²⁺ play a significant role in the aggregation mechanism of Aβ peptides. However, the nature of binding between metal and peptide has remained elusive; the detailed information on this from the experimental study is very difficult. Density functional theory (dft) (M06‐2X/6‐311++G (2df,2pd) +LANL2DZ) has employed to determine the force field resulting due to metal and histidine interaction. We performed 200 ns molecular dynamics (MD) simulation on Aβ_1‐42‐Zn²⁺, Aβ_1‐42‐Cu²⁺, and Aβ_1‐42‐Fe²⁺ systems in explicit water with different combination of coordinating residues including the three Histidine residues in the N‐terminal. The present investigation, the Aβ_1‐42‐Zn²⁺ system possess three turn conformations separated by coil structure. Zn²⁺ binding caused the loss of the helical structure of N‐terminal residues which transformed into the S‐shaped conformation. Zn²⁺ has reduced the coil and increases the turn content of the peptide compared with experimental study. On the other hand, the Cu²⁺ binds with peptide, β sheet formation is observed at the N‐terminal residues of the peptide. Fe²⁺ binding is to promote the formation of Glu22‐Lys28 salt‐bridge which stabilized the turn conformation in the Phe19‐Gly25 residues, subsequently β sheets were observed at His13‐Lys18 and Gly29‐Gly37 residues. The turn conformation facilitates the β sheets are arranged in parallel by enhancing the hydrophobic contact between Gly25 and Met35, Lys16 and Met35, Leu17 and Leu34, Val18 and Leu34 residues. The Fe²⁺ binding reduced the helix structure and increases the β sheet content in the peptide, which suggested, Fe²⁺ promotes the oligomerization by enhancing the peptide‐peptide interaction. Proteins 2016; 84:1257–1274. © 2016 Wiley Periodicals, Inc.     

Metal ion complexes of d-biotin in solution. Stability of the stereoselective thioether coordination

By spectrophotometry and ¹H nmr, several of the stability constants of the thioether complexes between Mg²⁺, Ca²⁺, Mn²⁺, Cu²⁺, Zn²⁺, Cd²⁺, or Ag⁺ and d-biotin (Bio), tetrahydrothiophene (Tht), and dimethyl sulfide (Dms) have been measured in 50% aqueous ethanol, 96% N,N-dimethylformamide (DMF), 98% d₆-dimethyl sulfoxide, or in D₂O. With decreasing concentration of water, the thioether interaction increases with the biologically important metal ions, whereas, e.g., Ag⁺ behaves in the opposite way. The stability of these complexes is, in general, quite small: for example, with d-biotin in 96% DMF (I = 1.0; 25°C) log K_M(Bio)^M = 0.03 and 1.64 for Cu²⁺ and Ag⁺, respectively; in D₂O (I = 0.5 for Ag⁺, all others 2–5; 27°C) log K_M(Bio)^M ? ?1.0, ?1.4, ?1.2, ?0.9, or 4.20 for Mg²⁺, Ca²⁺, Zn²⁺, Cd²⁺, or Ag⁺. In those cases where the difference log K_M(Tht)^M ? log K_M(Bio)^M can be calculated, it is in the order of 0.3 log units; this observation, as well as the chemical shifts measured, confirm the earlier suggestion that the interaction at the sulfur of biotin is stereoselective: the metal ion coordinates from “below” the tetrahydrothiophene ring of biotin to the sulfur atom, i.e., trans to the urea ring. It is emphasized that despite the low stability of these complexes with the biologically meaningful metal ions, the extent of the interaction is enough to create specific structures.     

Uptake of Metal Ions by Rhizopus arrhizus Biomass

Rhizopus arrhizus biomass was found to absorb a variety of different metal cations and anions but did not absorb alkali metal ions. The amount of uptake of the cations was directly related to ionic radii of La³⁺, Mn²⁺, Cu²⁺, Zn²⁺, Cd²⁺, Ba²⁺, Hg²⁺, Pb²⁺, UO₂²⁺, and Ag⁺. The uptake of all the cations is consistent with absorption of the metals by sites in the biomass containing phosphate, carboxylate, and other functional groups. The uptake of the molybdate and vanadate anions was strongly pH dependent, and it is proposed that the uptake mechanism involves electrostatic attraction to positively charged functional groups.