Inhibition of the biosynthesis ofN-acetylneuraminic acid by metal ions and seleniumin vitro

In liver homogenate the biosynthesis ofN-acetylneuraminic acid usingN-acetylglucosamine as precursor can be followed stepwise by applying different chromatographic procedures. In this cell-free system 16 metal ions (Zn²⁺, Mn²⁺, La³⁺, Co²⁺, Cu²⁺, Hg²⁺, VO ₃ ^– , Pb²⁺, Ce³⁺, Cd²⁺, Fe²⁺, Fe³⁺, Al³⁺, Sn²⁺, Cs⁺ and Li⁺) and the selenium compounds, selenium(IV) oxide and sodium selenite, have been checked with respect to their ability to influence a single or possible several steps of the biosynthesis ofN-acetylneuraminic acid. It could be shown that the following enzymes are sensitive to these metal ions (usually applied at a concentration of 1 mmoll^–1):N-acetylglucosamine kinase (inhibited by Zn²⁺ and vandate), UDP-N-acetylglucosamine-2-epimerase (inhibited by zn²⁺, Co²⁺, Cu²⁺, Hg²⁺, VO ₃ ^– , Pb²⁺, Cd²⁺, Fe³⁺, Cs⁺, Li⁺, selenium(IV) oxide and selenite), andN-acetylmannosamine kinase (inhibited by Zn²⁺, Cu²⁺, Cd²⁺, and Co²⁺). Dose dependent measurements have shown that Zn²⁺, Cu²⁺ and selenite are more efficient inhibitors of UDP-N-acetylglucosamine-2-epimerase than vanadate. As for theN-acetylmannosamine kinase inhibition, a decreasing inhibitory effect exists in the following order Zn²⁺, Cd²⁺, Co²⁺ and Cu²⁺. In contrast, La³⁺, Al³⁺ and Mn²⁺ (1 mmoll^–1) did not interfere with the biosynthesis ofN-acetylneuraminic acid. Thus, the conclusion that the inhibitory effect of the metal ions investigated cannot be regarded as simply unspecific is justified.Dedicated to Professor Theodor Günther on the occasion of his 60th birthday     

Contributions of cell wall and metal-binding peptide to Cd- and Cu-tolerances in suspension-cultured cells of tomato

Possible roles of cell wall and cytoplasmic peptides in the tolerance of cells to Cu²⁺ and Cd²⁺ ions were studied in suspension-cultured cells of tomato (Lycopersicon esculentum L. cv. Palace). Cu²⁺ and Cd²⁺ ions inhibited growth of wild type cells at concentrations more than 100 and 200 μM, respectively. Tomato cells readily developed tolerance to Cd²⁺ ions up to 1 mM but not to Cu²⁺ ions, after repeated subculturings in the presence of the respective ions. Such a metal-specific adaptation of cells was not due to the difference in the total uptakes between Cd²⁺ and Cu²⁺ ions by cells. Wild-type cells accumulated Cd²⁺ preferentially into the cytoplasmic peptide fraction and Cu²⁺ into the cell-wall fraction, when grown under the subtoxic metal conditions. Under excess metal conditions, Cd-tolerant cells produced greater amounts of Cd-binding peptides in the cytoplasm and retained lesser amounts of Cd²⁺ ions in the cell wall than did wild-type cells. In contrast, tomato cells grown in the presence of Cu²⁺ ions synthesized no detectable amounts of Cu-binding peptides in the cytoplasm and retained most of the Cu²⁺ in the cell-wall fraction, irrespective of cell lines. These results suggested that the cytoplasmic peptides rather than cell wall properties have a primary role in the response of tomato cells to excess metal environments.     

Layered structure of granules in upflow anaerobic sludge blanket reactor gives microbial populations resistance to metal ions

Metal ions (Cd²⁺, Cu²⁺, Ni²⁺, Zn²⁺ and Cr³⁺) did not affect glucose degradation or the production of methane during anaerobic digestion with intact and disintegrated granules from a UASB (Upflow Anaerobic Sludge Blanket) reactor. However, when Cu²⁺ was at 500 mg g^–1 VSS (volatile suspended solids) in the media, the glucose degradation rates and methane production rates decreased by 14% and 32% in disintegrated granules, respectively, whereas, in intact granules, decreases were 3% and 14%, respectively. When various electroplating metal ions were tested, 50% inhibition of acetate degradation and methane production were produced by 210–770 mg g^–1 VSS and 120–630 mg g^–1 VSS, respectively. The relative toxicity of the electroplating metals on methane production was in the order of Zn²⁺ (most toxic) > Ni²⁺ > Cu²⁺ > Cr³⁺ > Cd²⁺ (least toxic).     

Trace metals complexation behavior with root exudates induced by salinity from a mangrove plant Avicennia marina (Forsk.) Vierh

This study investigated the characteristics of exudates from mangrove plant Avicennia marina seedling roots under 0, 200 and 600?mM NaCl treatments and their complexation behavior with trace metals using excitation emission matrix (EEM) fluorescence spectrometry. Two fulvic-like fluorescence peaks, namely peak A (Em = 440?nm, Ex = 250?nm, UV fulvic-like compounds) and peak B (Em = 440?nm, Ex = 340?nm, visible fulvic-like compounds) were identified. The fluorescence intensities of peak A and peak B were enhanced by increasing salinity. Furthermore, the fluorescence of both peaks could be quenched by the ions of copper (Cu²⁺), manganese (Mn²⁺) and cadmium (Cd²⁺). Conditional stability constant (logK_a) exhibited that binding capacity of both peak A and peak B with trace metals are Cu²⁺?>?Mn²⁺?>?Cd²⁺ in the range from 2.21 to 4.01. Besides, Hill coefficient (n) >1 for Cu²⁺ but n?<?1 for Mn²⁺ and Cd²⁺. The results of high n and high logK_a for Cu²⁺ rather than Mn²⁺ and Cd²⁺ indicate that the fulvic-like compounds in root exudates of A. marina have maximum potential for Cu²⁺ complexation compared to Mn²⁺ and Cd²⁺, suggesting the fulvic acids in root exudates of A. marina have strong complexation with Cu²⁺ rather than Mn²⁺ and Cd²⁺.     

Melafen action on the growth of cyanobacteria and green microalgae cultured in stress conditions

Melafen stimulating effect on cell growth of cyanobacteria Synechococcus sp. PCC 6301 cultures amounted to 30–45% at 1000 lx illumination. The melafen effect decreased when cell cultures were exposed at the illumination of the saturation range (4000 lx). Growth rate and biomass increase of Anabaena variabilis, as well as the observed melafen stimulating effect, were higher on nitrogen-free medium compared to a nitrogen-containing one by 20–25%. We conclude that melafen activates photosynthetic processes and, probably, stimulates fixation of the atmospheric nitrogen in the cells. Opposite to the stimulating effect of melafen, ions of the heavy metal Cd²⁺ inhibited both biomass increase and the average number of the cells in the cyanobacteria A. variabilis colonies. The melafen added to the medium together with the Cd²⁺ ions decreased their negative effect. The other heavy metal ions, Cu²⁺, inhibited the growth of the cyanobacteria Synechococcus sp. PCC 6301 and green microalgae Chlorella vulgaris but had a stimulation effect on carbohydrate excretion by the cell cultures. Again, the melafen decreased the toxic effect of Cu²⁺ in this case. We suppose that melafen has an antistress activity at heavy metal ions presence and reduces their toxic effect on growth of phototrophic microorganisms.     

Effects of heavy metals on seed germination and early seedling growth of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Seed is a developmental stage that is highly protective against external stresses in the plant life cycle. In this study, we analyzed toxicity of essential (Cu²⁺ and Zn²⁺) and non-essential heavy metals (Hg²⁺, Pb²⁺ and Cd²⁺) on seed germination and seedling growth in the model species Arabidopsis. Our results show that seedling growth is more sensitive to heavy metals (Hg²⁺, Pb²⁺, Cu²⁺ and Zn²⁺) in comparison to seed germination, while Cd²⁺ is the exception that inhibited both of these processes at similar concentrations. To examine if toxicity of heavy metals is altered developmentally during germination, we incubated seeds with Hg²⁺ or Cd²⁺ only for a restricted period during germination. Hg²⁺ displayed relatively strong toxicity at period II (12–24 h after imbibition), while Cd²⁺ was more effective to inhibit germination at period I (0–12 h after imbibition) rather than at period II. The observed differences are likely to be due in part to selective uptake of different ions by the intact seed, because isolated embryos (without seed coat and endosperm) are more sensitive to both Hg²⁺ and Cd²⁺ at period I. We assessed interactive toxicity between heavy metals and non-toxic cations, and found that Ca²⁺ was able to partially restore the inhibition of seedling growth by Pb²⁺ and Zn²⁺.     

Citric acid fermentation from starch and dextrose syrups by a trace metal resistant mutant of Aspergillus niger

Potato starch and both untreated and decationized dextrose syrups were used as substrates for submerged citric acid biosynthesis using a mutant of Aspergillus niger. The same yield of product (80%) was achieved with both syrups and the starch despite having different trace metals content. The obtained mutant was more sensitive than the parent to Cd²⁺, Mo²⁺, and As³⁺, with decreasing yields of citric acid at 10 mg of ions l^–1. Fe²⁺, Mn²⁺, V²⁺ below 50 mg l^–1 and Cr³⁺, Ni²⁺, Cu²⁺ up to 100 mg l^–1, did not significantly inhibit citric acid production.     

Use of Saccharomyces cerevisiae for Cu2+ removal from solution: the advantages of using a flocculent strain

A flocculent strain of Saccharomyces cerevisiae S646-1B accumulated more Cu²⁺ (81 nmol mg^–1 dry wt) than the isogenic (except for the marker genes ade1 and trp1 and the gene FLO1) non-flocculent strain S646-8D (30 nmol mg^–1 dry wt), in the first 10 min of contact of the cells with Cu²⁺. Additionally, this strain flocculated in solutions of 0.2 mM Cu²⁺, Ni²⁺, Zn²⁺ and Cd²⁺. The potential of using flocculent strains in the bioremediation of heavy metals contaminated waste waters is discussed.     

Bacterial Degradation of EDTA

Degradation of EDTA (ethylenediaminetetraacetic acid) or metal–EDTA complexes by cell suspensions of the bacterial strain DSM 9103 was studied. The activity of EDTA degradation was the highest in the phase of active cell growth and decreased considerably in the stationary phase, after substrate depletion in the medium. Exponential-phase cells were incubated in HEPES buffer (pH 7.0) with 1 mM of uncomplexed EDTA or EDTA complexes with Mg²⁺, Ca²⁺, Mn²⁺, Pb²⁺, Co²⁺, Cd²⁺, Zn²⁺, Cu²⁺, or Fe³⁺. The metal–EDTA complexes (Me–EDTA) studied could be divided into three groups according to their degradability. EDTA complexes with stability constants K below 10¹⁶ (log K < 16), such as Mg–EDTA, Ca–EDTA, and Mn–EDTA, as well as uncomplexed EDTA, were degraded by the cell suspensions at a constant rate to completion within 5–10 h of incubation. Me–EDTA complexes with log K above 16 (Zn–EDTA, Co–EDTA, Pb–EDTA, and Cu–EDTA) were not completely degraded during a 24-h incubation, which was possibly due to the toxic effect of the metal ions released. No degradation of Cd–EDTA or Fe(III)–EDTA by cell suspensions of strain DSM 9103 was observed under the conditions studied.     

Mechanism of metal ion biosorption by fungal biomass

Alkali extracted mycelial biomass from Aspergillus niger, referred to as Biosorb, was found to sequester metal ions (Cd²⁺, Cu²⁺, Zn²⁺, Ni²⁺ and Co²⁺) efficiently both from dilute and concentrated solutions upto 10% of its weight (w/w). Sequestration of metal ions from a mixture was also efficient but with attendant antagonisms. The kinetics of metal binding by Biosorb indicated that it is a rapid process and about 70–80% of the metal is removed from solution in 5 min followed by a slower rate. The mechanism of metal binding is shown to be due to exchange of calcium and magnesium ions of the Biosorb during which equimolar concentrations of both the ions were released into the medium. Following this an efficient procedure for the regeneration and reuse of Biosorb was standardized by washing the biosorbent with calcium and magnesium solution (0.1 m). Biosorbents prepared from Neurospora, Fusarium and Penicillium also exhibited similar mechanisms for metal ion binding, though they had a lower metal binding capacity when compared with Biosorb. Chemical modification of carboxylic acid functional groups of the Biosorb resulted in loss of 90% of metal binding capacity which could not be restored even on regeneration. The significance of this finding on the metal sequestration mechanisms of microbial biosorbents is discussed.     

Functional comparison of metallothioneins MTT1 and MTT2 from Tetrahymena thermophila

Metallothioneins MTT1 and MTT2 from Tetrahymena thermophila have been characterized. The MTT1 contains mainly characteristic Cys-Cys-Cys and Cys-Cys clusters, but MTT2 contains mainly Cys-X-Cys cluster. Cd₁₆-MTT1 mainly consists of α-helix and β-turns, in contrast, Cd₁₁-MTT2 mainly consists of random coils. Reaction of Cd₁₆-MTT1 and Cd₁₁-MTT2 with nitric oxide leads to intramolecular disulfide bond formation, respectively. Binding stabilities of Cd²⁺, Hg²⁺ and Zn²⁺ to MTT1 are stronger than those to MTT2. Cu²⁺ can not replace Cd²⁺ from Cd₁₆-MTT1 complex, but can replace Cd²⁺ from Cd₁₁-MTT2 complex. The analysis of qRT-PCR revealed MTT2 mRNA levels were 31-fold higher than those of MTT1 under basal conditions. These results further suggest MTT1 possibly play a role in the detoxification of heavy metal ions, and MTT2 may be involved in the homeostasis of copper ions.     

Functional comparision between truncated MTT1 and truncated MTT2 from Tetrahyemna thermophila

Metallothioneins (MTs) are low-molecular-weight proteins with high Cys content and high metal-chelating ability. CdMT and CuMT subfamilies present different characteristics in Tetrahymena. To explore the effect of the cysteine arrangement and sequence length of MTs for binding different metal ions, MTT1, truncated MTT1 (TM1), MTT2, and truncated MTT2 (TM2) were expressed in E. coli. The half-maximal inhibiting concentrations (IC₅₀) of Cd²⁺ and Cu⁺ for the recombinant strains were different. Furthermore, E. coli cells expressing MTT1 and TM1 exhibited higher accumulating ability for Cd²⁺ than cells expressing MTT2 and TM2. However, the opposite is true for Cu⁺. The binding ability of the different recombinant proteins to Cd²⁺ and Cu⁺ were also different. MTT1 and truncated mutant TM1 were the preference for Cd²⁺, whereas MTT2 and truncated mutant TM2 were the preference for Cu⁺ coordination. These results showed that metal ion tolerance and accumulation ability not only depended on cysteine arrangement pattern but also on sequence length of MT in Tetrahymena.     

In vivo and in vitro effects of copper and cadmium on the plasma membrane H+-ATPase from cucumber (Cucumis sativus L.) and maize (Zea mays L.) roots

The plasmalemma vesicles isolated from cucumber and maize roots were used to study the effect of Cu²⁺ and Cd²⁺ on the hydrolytic and proton pumping activities of ATPase. In vivo application of metal ions to the plant growth solutions resulted in stimulation of the proton transport in maize. In cucumber roots the action of metals was not the same: cadmium stimulated the H⁺ transport through plasmalemma whereas Cu²⁺ almost completely inhibited it. Copper ions decreased the hydrolytic activity of H⁺-ATPase in cucumber, without any effect on this activity in membranes isolated from maize roots. The effect of cadmium on the hydrolytic activities was opposite: ATP-hydrolysis activity in plasmalemma was not altered in cucumber, whereas in maize its stimulation was observed. The amount of accumulated metals was not the main reason of different influence of metals on H⁺-ATPase activity in tested plants. In in vitro experiments Cu²⁺ inhibited H⁺ transport in the cucumber, to a higher degree than Cd²⁺ and both metals did not change this H⁺-ATPase activity of plasmalemma isolated from corn roots. Cu²⁺ added into the incubation medium reduced the hydrolytic activity of ATPase in the plasma membrane isolated from cucumber as well as from corn roots. Cd²⁺ diminished the hydrolytic activity of ATPase in cucumber, and no effect of Cd²⁺ in the plasmalemma isolated from corn roots was found. Our results indicated different in vitro and in vivo action of both metals on H⁺-ATPase and different response of this enzyme to Cu²⁺ and Cd²⁺ in maize and cucumber.     

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.     

Accumulation of Cd2+ and Pb2+ in the suspension cultures of Agave amaniensis and Costus speciosus and the determination of the culture's growth and phytosteroid content

Cell suspension cultures of Agave amaniensis and Costus speciosus were grown in media containing Cd^{2 +} up to 25 and 20 mg l^–1, respectively, and Pb²⁺ up to 40 mg l^–1. The cultures hyper-accumulated Cd²⁺ up to 900 and 530 g g^–1 and Pb²⁺ up to 1390 and 1170 g g^–1 dry wt. in their respective biomasses. Increasing Pb²⁺ up to 30 mg l^–1 increased the biomass production and total sitosterol content of Costus speciosus by up to 1.7- and 1.3-fold, respectively.     

Evidence for the Cd<Superscript>2+</Superscript> Activation of the Aryl Sulfatase from <Emphasis Type="Italic">Helix pomatia</Emphasis>

Often used to remove sulfate groups from carbohydrates, the regulatory properties of the aryl sulfatase from Helix pomatia remain little characterized. As many hydrolytic enzymes utilize exogenous metal ions in catalysis, the effect of various divalent metal ions on the sulfatase was investigated. Evidence for metal ion activation was collected, with Cd²⁺ being notable for effective activation. The enzyme was inhibited by Cu²⁺. The response of other common hydrolases to divalent metal ions was characterized. Activation by Cd²⁺ was not observed for chymotrypsin, rabbit liver esterase, or β-galactosidase. Instead, Cd was found to inhibit both the esterase and the galactosidase. Inhibition by Cu²⁺ and Zn²⁺ was also observed for some of these hydrolases.     

Hg2+-induced leakage of electrolytes and inhibition of NO3 − utilization inNostoc calcicola

Summary The effect of mercury (Hg²⁺) in the absence and presence of methylmercury (CH₃Hg⁺), cadmium (Cd²⁺), copper (Cu²⁺), nickel (Ni²⁺) and calcium (Ca²⁺) on Nostoc calcicola Bréb. has been studied in terms of electrolyte leakage, NO₃ ^– uptake and in vivo nitrate reductase (NR) activity to discover any possible correlation among such parameters under Hg²⁺ stress. Leakage of electrolytes from Hg²⁺-treated cyanobacterial cells was directly proportional to Hg²⁺ concentrations and exposure time. In comparison to NO₃ ^– uptake, an about 60-fold slower rate of NR activity was observed in the untreated cultures, the former being five times more Hg²⁺-sensitive. A non-competitive synergistic interaction of Hg²⁺ with CH₃Hg⁺ or Cd²⁺ and antagonistic with that of Ni²⁺ or Ca²⁺ has been observed for both the processes of NO₃ ^– utilization. The antagonistic interaction of Cu²⁺ with Hg²⁺ in terms of NO₃ ^– uptake and synergistic with respect to NR activity, has been attributed to the dual bonding preference of Cu²⁺ for cellular ligands. These findings suggest that (a) a statistically significant correlation exists among such parameters; (b) Hg²⁺ predominantly attacks the cyanobacterial cell membrane; (c) Hg²⁺ inhibits NO₃ ^– utilization; (d) the presence of other cations increases or decreases the inhibitory actions of Hg²⁺.     

Improvement of indole alkaloid production in Catharanthus roseus cell cultures by osmotic shock

Catharanthine production in Catharanthus roseussuspension cell cultures was increased by about 4-fold to 28 mg l^–1, 23 mg l^–1and 24 mg l^–1by adding sodium alginate, mannitol or polyvinyl pyrrolidone, respectively. Sodium alginate and polyvinyl pyrrolidone also enhanced ajmalicine production to 28 mg l^–1and 31 mg l^–1, respectively. Up to 55–70% of the total alkaloids were released into the medium. These treatments could stimulate higher alkaloid production in C. roseuscell cultures than NaCl and KCl stresses. The possible mechanisms for these treatment effects are discussed.     

Adsorption of Cu(II), Cd(II), and Pb(II) from aqueous single metal solutions by mercerized cellulose and mercerized sugarcane bagasse chemically modified with EDTA dianhydride (EDTAD)

This work describes the preparation of new chelating materials derived from cellulose and sugarcane bagasse for adsorption of Cu²⁺, Cd²⁺, and Pb²⁺ ions from aqueous solutions. The first part involved the mercerization treatment of cellulose and sugarcane bagasse with NaOH 5 mol/L. Non- and mercerized cellulose and sugarcane bagasse were then reacted with ethylenediaminetetraacetic dianhydride (EDTAD) in order to prepare different chelating materials. These materials were characterized by mass percent gain, X-ray diffraction, FTIR, and elemental analysis. The second part consisted of evaluating the adsorption capacity of these modified materials for Cu²⁺, Cd²⁺, and Pb²⁺ ions from aqueous single metal solutions, whose concentration was determined by atomic absorption spectroscopy. These materials showed maximum adsorption capacities for Cu²⁺, Cd²⁺, and Pb²⁺ ions ranging from 38.8 to 92.6 mg/g, 87.7 to 149.0 mg/g, and 192.0 to 333.0 mg/g, respectively. The modified mercerized materials showed larger maximum adsorption capacities than modified non-mercerized materials.     

Isolation and Molecular Characterization of Heavy Metal-Resistant Azotobacter chroococcum from Agricultural Soil and Their Potential Application in Bioremediation

Pollution of soil with heavy metals, herbicides, antibiotics and other chemicals is known to have a negative effect on microbial activities. Therefore, the aim of this study was to isolate cultures of Azotobacter sp. from polluted and unpolluted soils and to study the effect of these pollutants on their growth. A total of 120 Azotobacter sp. were isolated from soils irrigated with wastewater (contaminated soils) and groundwater (uncontaminated soils). These isolates were screened for resistance to heavy metals, herbicide and antibiotics. Also, the soils from which the cultures were isolated were analyzed for the concentrations of Zn²⁺, Cd²⁺, Cu²⁺, Pb²⁺ and Mn²⁺ they contained. Contaminated soil showed high levels of heavy metals as compared to uncontaminated soil. The size of the Azotobacter population in contaminated soil was lower than that in uncontaminated soil. Of the Azotobacter isolates, 64 that were recovered from contaminated soil exhibited high resistance to heavy metals (Hg²⁺, Cd²⁺, Cu²⁺, Cr³⁺, Co²⁺, Ni²⁺, Zn²⁺ and Pb²⁺) and herbicide 2,4-D compared to 56 isolates from uncontaminated soil. Also, isolates from contaminated soil showed high resistance to chloramphenicol, nitrofurantoin and co-trimoxazole compared to those isolated from uncontaminated soil. The majority of Azotobacter isolates from contaminated soil showed multiple-resistance to different metal ions and antibiotics. All isolates failed to grow at pH less than 6. Salt concentration (5%) was found to be inhibitory to all isolates. The most potent isolates from contaminated soil that showed multiresistance to all substances tested were identified on the basis of morphological and biochemical characteristics, and 16S rRNA as A. chroococcum. These resistant isolates could be employed in contaminated soils and/or bioremediation.