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1.
Zinc is essential for normal growth, development and brain function although little is known about brain zinc homeostasis. Therefore, in this investigation we have studied65Zn uptake from blood into brain and other tissues and have measured the blood-brain barrier permeability to65Zn in the anaesthetized rat in vivo. Adult male Wistar within the weight range 500–600 g were used.65ZnCl2 and [125I]albumin, the latter serving as a vascular marker, were injected in a bolus of normal saline I.V. Sequential arterial blood samples were taken during experiments that lasted between 5 min and 5 hr. At termination, samples from the liver, spleen, pancreas, lung, heart, muscle, kidney, bone, testis, ileum, blood cells, csf, and whole brain were taken and analysed for radio-isotope activity. Data have been analysed by Graphical Analysis which suggests65Zn uptake from blood by all tissues sampled was unidirectional during this experimental period except brain, where at circulation times<30 min,65Zn fluxes were bidirectional. In addition to the blood space, the brain appears to contain a rapidly exchanging compartment(s) for65Zn of about 4 ml/100g which is not csf.  相似文献   

2.
Summary The Zn contents of twenty-nine alluvial soils from Egypt were chemically fractionated into: water soluble+exchangeable, weakly bound to inorganic sites, organically bound, occluded as free oxide material, and residual mainly in the mineral structure. On the average these fractions constituted about 0.01, 1.20, 28.6, 21.5 and 45.5% of the total soil Zn respectively which averaged 76.25 ppm. Significant correlations were obtained between each individual Zn-fraction and some soil variable.Zinc adsorption isotherms were developed for seven soils suspended in dilute ZnCl2 solution in the presence of either 0.05M CaCl2 solution (Specific adsorption) or deionized water (Total adsorption). The Langmuir constants (adsorption maximum and bonding energy) were calculated. The average value of specific adsorption maximum was 1.94 mg Zn/g soil and of total adsorption maxima was 11.54 mg Zn/g soil. Correlation analysis showed that CEC, free Fe2O3 and clay content were the dominant soil variables contributing towards specific Zn adsorption. The (Zn) (OH)2 ion concentration products in the solutions when Zn adsorption corresponded to the Langmuir adsorption maxima were 0.92×10–17 in the specific adsorption treatment, and 1.35×10–15 in the total adsorption treatment. These values are within the solubility range of Zn (OH)2 and ZnCO3. The values of Langmuir bonding energy constants showed that Zn was more strongly adsorbed by low carbonate or carbonate-free soils than by carbonate-rich soils.  相似文献   

3.
Summary Zinc adsorption was studied in suspensions of six soils of different physicochemical characteristics in dilute ZnSO4 solutions. At low concentrations, Zn2+ adsorption was described by the Langmuir adsorption equation. The calculated Langmuir adsorption maxima were related positively to clay and carbonate content and negatively with organic matter content of soils. Multiple regression analysis revealed that zinc adsorption maxima can be predicted with good precision from information in soil survey reports. When the added Zn2+ exceeded the adsorption maximum, the solid phase of zinc controlling its concentration in solution was either zinc hydroxide or carbonate so long as soil carbonates were present. The values of zinc potential also indicated that soils retain Zn2+ more strongly than Zn(OH)2 or carbonate. Postgraduate student Professor of Soils. Professor of Soils.  相似文献   

4.
A sequential extraction scheme was combined with sorption isotherm analysis in order to investigate sorption of sewage sludge-derived Cu and Zn to the A-horizon of a humic-gley soil as a whole, and to the operationally defined exchangeable (1?M MgCl2), carbonate (1?M NaOAc), Fe/Mn oxide (0.04?M NH2OH.HCl), and organic (0.02?M HNO3+30% H2O2) soil fractions. Sorption parameters were compared for a sample of sludge leachate (with 97.4% of Cu and 63.2% of Zn present as dissolved metal-organic matter complexes, as calculated by geochemical modeling involving MINTEQA2 and verified using an ion exchange resin method) with that of a reference solution exhibiting the same chemical characteristics as the leachate, except for the presence of dissolved organic material. Dissolved metal-organic matter complexes were found to significantly (P<0.05) depress sorption to the bulk soil and each fraction. The greatest depression of Cu and Zn sorption was observed for the exchangeable, carbonate, and Fe/Mn oxide fractions, while the organic fraction of the soil was the least affected. This reflects a greater affinity for the exchangeable, carbonate, and Fe/Mn oxide fractions by the free divalent metal (Cu2+, Zn2+), with sorption by these fractions attributed to cation exchange, chemisorption, and co-precipitation processes. The sorption characteristics of the organic fraction indicated that Cu and Zn sorption by soil organic matter mostly involved dissolved metal-organic matter complexes. This may be attributed to hydrophobic interactions between nonpolar regions of the dissolved metal-organic matter complexes and solid-phase soil organic matter.  相似文献   

5.
Summary Laboratory incubation experiment was conducted with a clay loam alluvial lowland rice soil to study the relative effectiveness of two sources of Zn (ZnEDTA and ZnSO4) in maintaining Zn availability in soil under two moisture regimes (saturated and waterlogged) both in presence as well as absence of added organic matter. The results showed that ZnEDTA was always more effective than ZnSO4 in maintaining higher amount of zinc in available form in soil for a longer perid. Results of greenhouse experiment conducted with rice showed that concentration and uptake of Zn by roots were generally higher with ZnEDTA than with ZnSO4 both in presence and absence of added organic matter, whereas in respect of shoot this was true only in absence of added organic matter.  相似文献   

6.
This study was designed to determine the effect of zinc on the biological half-lives of 65Zn in whole body and liver and on distribution of 65Zn in different organs of rats following nickel toxicity. Sprague-Dawley (SD) rats received either nickel in the form NiSO4·6H2O at a dose of 800 mg/L in drinking water, zinc in the form of ZnSO4·7H2O at a dose of 227 mg/L in drinking water, and nickel plus zinc or drinking water alone for a total duration of 8 wk. All of the rats were injected with a tracer dose of 0.37 MBq 65Zn at the end of the treatment period. The effects of different treatments were studied on biological half-lives of 65Zn in whole body and liver and on the distribution of 65Zn in different organs of rats. In the present study, we have noted that nickel treatment to normal rats caused a significant decrease in the slow component (Tb2) in liver, which improved following zinc supplementation. Nickel administration to normal-diet-fed animals caused significant lowering in the percentage uptake of 65Zn values in the brain, liver, and intestine. However, the administration of zinc to nickel-treated rats improved the status of 65Zn in different organs. The Tb2 in the liver and the percentage uptake of 65Zn values elevated following zinc supplementation to nickel-treated rats.  相似文献   

7.
Mineralization and redistribution of carbon from14C-labelled oat shoots and [14C(U)] labelled glucose, leucine, acetate and phenylacetate were studied in light loamy sand and medium clay loam under different levels of mineral nutrition. Losses of mineralized14C as CO2 were greater in the sandy soil than in the clay soil. NPK and NPK+Ca fertilization increased the rates of decay of the introduced plant organic matter. Among the small molecular organic compounds glucose was degraded fastest and phenylacetate slowest. Incorporation of radioactive carbon into humus fractions varied and depended on the nature of the compound introduced and on the soil type. Carbon of glucose, phenylacetate and acetate was mainly incorporated into fulvic acids, whereas14C of leucine was almost evenly distributed between humic and fulvic acids and14C of oat residues in fulvic acids and humin fractions. There was significantly higher incorporation of14C into humic acids and lower incorporation into humins in the sandy soil compared to the clay soil. NPK+Ca decreased the conversion of14C from phenylacetate and acetate to bitumens and increased its content in humic acids, particularly in the clay soil. The incorporation of14C from phenylacetate to humins benefitted from mineral fertilization during the first 30 days of the experiment in both soils.  相似文献   

8.
Summary The chemical equilibrium and E value for Zn was calculated in 9 soils of Haryana (India) which represented mainly entisol and inceptisols. The rate of isotopic exchange between65Zn and native soil Zn was quite rapid and radioactivity reduced to about 0.2 per cent of initial activity after 3 days but total Zn concentration in soil solution did not decrease. In 5 out of 9 soils equilibrium was established in 2 to 3 days. In typic ustochrepts having high clay content (3,4) equilibrium was attained in two days. In typic udipsamments (2), typic camborthids (8) and aquic vertic ustichrepts (9) it took 3 days to set equilibrium. Typic ustochrepts (5) took 5 days for equilibrium whereas typic ustipssaments (6) and typic camborthids of high O.C. did not attain equilibrium even in 7 days. This indicated that the chemistry and availability of Zn in soil would depend on soil types. When ZnE was estimated by applying activity after equilibrium with carrier Zn and by applying activity with carrier Zn before equilibrium was set, there was no agreement in ZnE in two methods. Increasing ZnE with increasing Zn dose was observed by both methods only in alkaline typic ustochrept (4). In some soils higher E values than added amounts were observed, whereas, in soil 8 negative E values were obtained. The E values become erratic and are over estimated where complex reactions take place due to high pH, high O.C. and high complex forming elements.  相似文献   

9.
Zinc homeostasis was investigated in Nostoc punctiforme. Cell tolerance to Zn2+ over 14 days showed that ZnCl2 levels above 22 μM significantly reduced cell viability. After 3 days in 22 μM ZnCl2, ca. 12% of the Zn2+ was in an EDTA-resistant component, suggesting an intracellular localization. Zinquin fluorescence was detected within cells exposed to concentrations up to 37 μM relative to 0 μM treatment. Radiolabeled 65Zn showed Zn2+ uptake increased over a 3-day period, while efflux occurred more rapidly within a 3-h time period. Four putative genes involved in Zn2+ uptake and efflux in N. punctiforme were identified: (i) the predicted Co/Zn/Cd cation transporter, putative CDF; (ii) the predicted divalent heavy-metal cation transporter, putative Zip; (iii) the ATPase component and Fe/Zn uptake regulation protein, putative Fur; and (iv) an ABC-type Mn/Zn transport system, putative zinc ZnuC, ZnuABC system component. Quantitative real-time PCR indicated the responsiveness of all four genes to 22 μM ZnCl2 within 3 h, followed by a reduction to below basal levels after 24 h by putative ZIP, ZnuC, and Fur and a reduction to below basal level after 72 h by putative CDF efflux gene. These results demonstrate differential regulation of zinc transporters over time, indicating a role for them in zinc homeostasis in N. punctiforme.  相似文献   

10.
The objectives of the present paper were: (i) to determine isotopically exchangeable zinc using two isotopic exchange methods (E and L values) in a series of polluted and non-polluted Swiss agricultural soils, and (ii) to evaluate the ability of chemical extraction methods to estimate plant-available soil Zn using isotopic techniques. The surface horizon (0–20 cm) of seven polluted and non-polluted soils representing a wide range in physico-chemical properties and Zn contents were sampled. An isotopic exchange kinetics (IEK) approach was used to assess, in a batch experiment, the isotopically exchangeable Zn content (E value). In order to determine the L values, a pot experiment was carried out with Lolium multiflorum (cv. Axis) in a growth chamber using a 65Zn-isotope dilution technique. Total Zn uptake and the isotopic composition (65Zn/stableZn) were determined in Lolium multiflorum for five successive cuts. The amounts of zinc extracted by different chemicals were compared with L values and regression parameters were estimated. The isotopic composition in soil extracted by DTPA and EDTAAc at the end of the pot experiment was also determined. Results showed that the equation describing the increase of isotopically exchangeable Zn with time could be extrapolated to three months for polluted and non-polluted neutral and acidic soils, and that the results were not different from the amount of isotopically exchangeable Zn experimentally determined with Lolium multiflorum (L value). In alkaline soils however, results suggest that either 65Zn sorption occurred in the batch experiment or that the concentration of Zn in the soil solution had been overestimated, leading to an overestimation of the E value compared to the L values. Furthermore, the specific activities measured in DTPA and EDTA extractions at the end of the pot experiment were significantly different compared to the specific activity of the plant, showing that both these chelating agents extract neither all the available soil Zn nor only the available soil Zn for plants. Abbreviations: C Zn– concentration of Zn in a soil water extract (mg Zn L?1); C Zn?Plant– concentration of Zn in plant shoots (mg Zn kg?1 DM); DTPA – diethylene triamine pentaacetic acid; E 1\min– amount of Zn isotopically exchangeable within one min (mg Zn kg?1 soil); E (t)\exp– amount of Zn isotopically exchangeable after t min derived from experimental results (mg Zn kg?1 soil); E (t)pred– amount of Zn isotopically exchangeable after t min predicted using kinetic parameters derived from a 100 min long isotope exchange kinetic experiment together with C Zn, and ZnHNO3 (mg Zn kg?1 soil); EDTA – ethylene diamine tetraacetic acid; ICP– isotopic composition of Zn in plant shoots; ICDTPA– isotopic composition of Zn in the soil DTPA extract; ICEDTA– isotopic composition of Zn in the soil EDTA extract; ICSE– isotopic composition of Zn in the soil extracts; IEK – isotope exchange kinetics; L value – amount of plant available Zn (mg Zn kg?1 soil); Lolium multiflorum; TEA – Triethanolamine; ZnDTPA– Zn extractable by 0.005 M DTPA + 0.01 M CaCl2 + 0.1 M TEA; ZnEDTA?NH4Ac– Zn extractable by 0.5 M NH4Ac, 0.02 M EDTA; ZnEDTA?Ca(NO3)2– Zn extractable by 0.005 M EDTA, 0.01 M Ca(NO3)2; ZnKCl– Zn extractable by 1 M KCl; ZnCaCl2– Zn extractable by 0.01 M CaCl2; ZnNaNO3– Zn extractable by 0.1 M NaNO3; ZnHNO3– Zn extractable by 2 M HNO3.  相似文献   

11.
Distribution and retention of zinc in the presence of cadmium and copper was studied in rats exposed repeatedly to these metals. The experiment was performed on white rats of the Wistar strain. The animals were divided into four groups/five rats each: 1)65ZnCl2; 2)65ZnCl2+CdCl2; 3)65ZnCl2+CuCl2; and 4) control group. Rats were administered sc every other day for two weeks:65ZnCl2−5 mg Zn/kg; CdCl2−0,3 Cd/kg; and CuCl2−2 mg Cu/kg. The zinc content was measured in rat tissues by γ-counting. Effect of Cd and Cu on subcellular distribution of zinc in the kidney and liver and on the level of metallothionein were also examined. Whole body retention of zinc under the influence of cadmium was lower than that observed in animals treated with zinc alone. However, copper increased twofold the whole body retention of zinc. Cadmium elevated the accumulation of zinc only in the kidneys nuclear fraction and liver soluble fraction. In the kidneys and liver, copper elevated the accumulation of zinc, in the nuclear, mitochondrial, and soluble fractions. The level of metallothionein-like proteins (MT) in the kidneys after a combined supply of zinc and copper was significantly increased with respect to the group of animals treated with zinc alone. These results indicated complex interactions between cadmium, copper, and zinc that can affect the metabolism of each of the metals.  相似文献   

12.
Summary Greenhouse and laboratory studies were conducted to study the effect of zinc sources and methods of application on correcting zinc deficiency in flooded rice grown on Vertisol from Ngala, northern Nigeria, using the variety IR-20.Plant dry matter yield was similar for ZnSO4, ZnEDTA, metallic Zn and fritted Zn with mixed soil application. Zinc uptake was affected in the following order; ZnSO4 > ZnEDTA > metallic Zn > fritted Zn. Comparable dry matter yield and zinc uptake were obtained with mixing, surface broadcasting and banding of ZnEDTA. Mixing the fritted Zn gave higher dry matter yield and zinc uptake than broadcasting or banding.Seed soaking with a suspension of fritted Zn resulted in higher dry matter yield and zinc uptake than with ZnEDTA solution. Seed soaking for 24 hours with fritted Zn suspension at a concentration of about 0.5 per cent Zn appeared to be a suitable method for applying zinc with direct seeded rice.  相似文献   

13.
Zinc is essential for normal development and function of the CNS although much is to be learned about brain Zn homeostasis. In these experiments adult male Wistar rats within the weight range 500–600 g were used. Ventriculo-cisternal perfusion was performed to allow the measurement of65Zn fluxes between blood and csf across the choroid plexuses. Blood-brain or blood-cerebrospinal fluid barrier permeability to65Zn has been determined by graphical analysis in experiments that lasted between 5 and 180 minutes. Cerebral capillary permeability to65Zn was found to be low with a Kin of about 5×10–4ml/min/g. Choroid plexus permeability to65Zn was about 12 fold greater, although Zn influx to brain via this route was <5% that across cerebral capillaries. The autoradiographic distribution of65Zn in brain showed regional variation with lowest levels in white matter and high levels in the dentate gyrus and hippocampus.  相似文献   

14.
A series of mono- and dinuclear zinc complexes of 3,6,9,17,20,23-hexaaza-29,30-dihydroxy-13,27-dimethyl-tricyclo[23,3,111,15]triaconta-1(28),11,13,15(30),25,26-hexaene (H2L or BDBPH) have been defined in solution by potentiometry. The crystal structure of [Zn2C26H40N6O2(CH3OH)2]·Br2 has been determined by X-ray. Each zinc ion is coordinated to three nitrogen atoms, a bridged-phenolic oxygen atom, and a methanolic oxygen atom, which define a six-coordinated octahedron. Bond lengths of ZnN are in the range of 2.104(3)-2.120(3) Å and distances between Zn and O (bridged-phenolic oxygen) are 2.052(2), 2.062(2) Å, respectively. The dinuclear complexes: [Zn2L]2+ and [Zn2L(OH)]+ play crucial roles in hydrolytic reaction of tris(4-nitrophenyl)phosphate. A possible mechanism showed that [Zn2L(OH)]+ acts as a nucleophile and [Zn2L]2+ stabilizes the formation of the intermediate: [Zn2L-BNP].  相似文献   

15.
Solubility of metal in contaminated soils is a key factor which controls the phytoavailability and toxic effects of metals on soil environment. The chemical equilibria of metal ions between soil solution and solid phases govern the solubility of metals in soil. Hence, an attempt was made to identify the probable solid phases (minerals), which govern the solubility of Zn2+ and Cd2+ in zinc smelter effluent-irrigated soils. Estimation of free ion activities of Zn2+ (pZn2+) and Cd2+ (pCd2+) by Baker soil test indicated that metal ion activities were higher in smelter effluent-irrigated soils as compared to that in tubewell water-irrigated soils. Identification of solid phases further reveals that free ion activity of Zn2+ and Cd2+ in soil highly contaminated with Zn and Cd due to long-term irrigation with zinc smelter effluent is limited by the solubility of willemite (Zn2SiO4) in equilibrium with quartz and octavite (CdCO3), respectively. However, in case of tubewell water-irrigated soil, franklinite (ZnFe2O4) in equilibrium with soil-Fe and exchangeable Cd are likely to govern the activity of Zn2+ and Cd2+ in soil solution, respectively. Formation of highly soluble minerals namely, willemite and octavite indicates the potential ecological risk of Zn and Cd, respectively in smelter effluent irrigated soil.  相似文献   

16.
Zinc (Zn) is recognized as an essential nutrient, and is added as a supplement to animal and human diets. There are claims that zinc methionine (ZnMet) forms a stable complex that is preferentially transported into tissues, and this has contributed to uncertainty about conflicting reports on the bioavailability of various Zn compounds. This study evaluated the cellular and intestinal uptake of inorganic and organic forms of Zn. Steady-state uptake of65Zn by human intestine epithelial cells, and monkey kidney fibroblasts was not significantly different with zinc chloride (ZnCl2), ZnMet, or zinc propionate (ZnProp) (P > 0.05). Uptake of65Zn from zinc chelated with EDTA was significantly lower (P < 0.01). In live mice,65Zn uptake by perfused intestine and deposition in intestine and liver showed no significant difference between ZnCl2 and ZnMet. Equimolar [65Zn]methionine and zinc[35S]methionine were prepared according to a patented method that yields “ complexed” Zn. Cellular uptake of the radiolabeled methionine was <0.1% of the radiolabeled Zn from these complexes, indicating separate uptake of the Zn and methionine. Gel filtration did not distinguish between65Zn in ZnCl2, ZnProp, or reagent ZnMet, though feed-grade ZnMet containing >10% protein did give a higher-mol-wt form of65Zn. Results of this study show equivalent uptake of Zn from inorganic and organic compounds, and support recent feed trials on Zn bioavailability.  相似文献   

17.
Retention, dynamics of75Se and65Zn distribution, and elimination were studied in rats after separate or joint single doses of these metals. White female Wistar rats were divided into four groups (fifteen rats each). Group I received Na2 75SeO3 (0.1 mg Se/kg i.g.), group II received Na2 75SeO3+ZnCl2 (5 mg Zn/kg s.c.), group III received65ZnCl2, and group IV received65ZnCl2+Na2SeO3. The zinc and selenium contents in the tissues were estimated during 120 h after administration; excretion in urine and feces of animals was determined throughout the experiment. Combined administration of zinc and selenium resulted in an enhanced selenium retention in the brain, spleen, kidneys, blood, lungs, and heart. A selenium-induced increase in the concentration of zinc was noted in the bowels, blood, liver, kidneys, spleen, brain, and lungs. The effects of the zinc/selenium interaction were visible especially in the lowered level of excretion of these elements. Zinc induced a decrease in the excretion of selenium in urine, with no concomitant changes in the excretion in feces. However, a visible decrease in the excretion of zinc in the feces was observed in the presence of selenium. The present results indicate an occurrence of clear-cut interaction effects between zinc and selenium administered simultaneously in the rat.  相似文献   

18.
Summary The uptake of Zn65 by seeds ofVicia faba, soaked in a Zn65Cl2 solution and maintained within the experimental condition described in the text, was constant and did not depend on the amount of the radionuclide given. However, the percentage of the total absorbed Zn65 retained by seed coats varied according to the cultural variety ofVicia faba. Most of the Zn65 entering the seed, remained bound to the seed coat, the remainder entered the embryo and cotyledons and was concentrated around the abaxial part of this latter, lessening towards the adaxial region. The fate of the isotope was followed in all stage of plant development. Its distribution was represented as counts per minute per unit mass (dry) of plant tissue, and on a percentage basis. In general, zinc decreased in the main root acropetally then increased at the root stem transition zone, decreased again in the first internode than increased acropetally towards the apex of the stem. Anthers had the highest counts per unit mass compared to other plant organs, including the rest of the flower. Zn65 distribution calculated on a percentage basis increased in that order: Roots, stem, leaves in one month old plants and older. These observations held also in plants grown from —Zn medium and from seeds to which radioactive zinc was introduced before and during the formation of their pods. Water-soluble Zn65 was also investigated in different plant's organs. Some of these results are compared and discussed with those previously obtained using a histochemical method.  相似文献   

19.
Copper,Lead, Cadmium,and Zinc Sorption By Waterlogged and Air-Dry Soil   总被引:1,自引:0,他引:1  
Competitive sorption of copper (Cu), lead (Pb), cadmium (Cd), and zinc (Zn) was studied in three soils of contrasting chemical and physical properties under air-dry and waterlogged conditions. Competitive sorption was determined using the standard batch technique using six solutions, each with Cu, Pb, Cd, and Zn concentrations of approximately 0, 2.5, 5, 10, 20, and 50?mg L?1Waterlogged soils tended to sorb higher amounts of added Cu, Pb, Zn and Cd relative to soils in the air-dry condition; however, this increase in sorption was generally not statistically (p<0.05) significant. The magnitude of sorption under both waterlogged and air-dry conditions was affected by the type and amount of soil materials involved in metal sorption processes, and competition between other metals for the sorption sites. Metal sorption was closely correlated with soil properties such as cation exchange capacity, organic carbon, and Fe and Mn hydrous oxides. Exchangeable Al may have markedly reduced metal sorption due to its strong affinity for the sorption sites, while increases in exchangeable Mn may have enhanced Zn and Cd sorption. Heavy metal sorption was best described as a combination of both specific and nonspecific interactions. The extractability of Cu, Pb, Cd, and Zn under waterlogged and air-dry conditions was also studied. Three solutions containing these metals were mixed with each soil to achieve a final concentration of 0, 50, and 500?mg kg?1. Each soil was extracted every 7 days using 1?M MgCl2 (pH 7) to determine metal extractability. Metal extractability initially decreased then increased due to waterlogging. The increased extractability of added metals was closely related to increased solubility of Fe and Mn suggesting that dissolution of Fe and Mn, oxides under reducing conditions caused a release of previously sorbed Cu, Pb, Cd, and Zn.  相似文献   

20.
Phosphotriesterase from Pseudomonas diminuta (PTE; EC 3.1.8.1) hydrolyzes organophosphate insecticides and chemical warfare agents. The two zinc cations in the active center can be substituted. Co2+-containing PTE is the most efficient but least stable isoform. Gel filtration showed that PTE is monomeric at the submicromolar concentrations used in kinetic assays. The analysis of the recombinant enzyme by X-ray fluorescence spectrometry and CCT-ICP-MS, confirms that recombinant Zn-PTE contains only Zn2+ whereas Co-PTE has Zn2+ and Co2+ in equimolar amount, with Co2+ most likely in the reported labile β-site. We noted that recombinant PTE is unstable at low concentrations and must be stabilized by a protein environment. We tested the effect of excess of various metal cofactors on PTE-catalyzed hydrolysis of paraoxon. We notably observed that ZnCl2 induces a non-competitive partial inhibition of Zn2+- and Co2+-PTE at pH 8.5 (apparent Ki=155 μM and 52 μM, respectively). Inhibition results from interactions with colloidal Zn(OH)2 formed in alkaline buffer that alters the catalytic machinery. NiCl2 caused a similar effect at higher concentrations (apparent Ki=3 mM). We observed that mutating His123, a surface residue close to an alleged allosteric site, dramatically altered the bacterial expression yield of Co2+-PTE, Ki for Zn(OH)2 inhibition, kcat (up to 60 fold) for paraoxon hydrolysis, but not KM. Issues addressed in this work are important for future biotechnological developments of PTE as a detoxifying enzyme.  相似文献   

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