真菌还原Cr（VI）的研究

从不同来源的样品中分离筛选出几株抗Ｃｒ（ＶＩ）的真菌,他们能在含３００￣５００ｍｇ／ＬＫ２Ｃｒ２Ｏ７的蔗糖合成培养基中生长,其中ＢＳ－１菌株抗Ｋ２Ｃｒ２Ｏ７达９００ｍｇ／Ｌ．ＢＳ－１等４株真菌在含２００ｍｇ／Ｌ Ｋ２Ｃｒ２Ｏ７的培养基中生长４￣６ｄ后,培养液中的Ｃｒ（ＶＩ）已全部消失。这些真菌经鉴定为青霉菌ＢＳ－１和ＢＳ－３,黑曲霉ＢＲ－４和黄曲霉ＢＸ－１。经紫外可见光扫描及化学分析证实,高毒的Ｃ     

Chromium(VI) bioaccumulation capacities of adapted mixed cultures isolated from industrial saline wastewaters

Enrichment mixed cultures tolerating relatively high concentrations of chromium and salt ions were isolated and their bioaccumulation properties improved by adaptation. Mixed cultures were enriched in Nutrient Broth media containing 25-300 mg l(-1) Cr(VI) and 0%, 2%, 4%, 6% (w/v) NaCl. Bioaccumulation of Cr(VI) was studied in a batch system as a function of initial pH (7, 8 and 9), Cr(VI) and NaCl concentrations. Increasing NaCl and Cr(VI) concentrations led to significant decreases in percentage uptake and dried weight of mixed cultures but increased maximum specific chromium uptake. The maximum specific chromium uptake value at pH 8 was 58.9 mg g(-1) for 316.1 mg l(-1) Cr(VI) in the absence of NaCl, while at pH 9 it was 130.1 mg g(-1) in media including 194.5 mg l(-1) Cr(VI) and 2% NaCl concentrations. At 4% NaCl, the maximum Cr(VI) uptake of 127.0 mg g(-1) for 221.1 mg l(-1) Cr(VI) occurred at pH 9, while at 6% NaCl the maximum Cr(VI) uptake of 114.9 mg g(-1) for 278.1 mg l(-1) Cr(VI) was found at pH 7.     

Biological removal of carcinogenic chromium(VI) using mixed Pseudomonas strains

The contamination of soil and wastewaters with Cr(VI) is a major problem. It has been suggested that microbial methods for Cr(VI) reduction are better than chemical methods, as they do not add other ions or toxic chemicals to the environment. In this study an aerobic reduction of Cr(VI) to Cr(III) by employing mixed Pseudomonas cultures isolated from a marshy land has been reported. The role of chromium concentration, temperature, pH and additives on the microbial reduction of Cr(VI) has been investigated. NADH was found to enhance the rate of reduction of Cr(VI). Complete reduction of chromium(VI) has been possible even at chromium(VI) concentrations of 300 ppm. Ions like SO(4)(2-) and poly-phenols inhibited the metabolic activity relating to Cr(VI) reduction. Under optimal conditions 100 mg/L of Cr(VI) was completely reduced within 180 min.     

Environmental and Kinetic Parameters for Cr(VI) Bioreduction by a Bacterial Monoculture Purified from Cr(VI)-Resistant Consortium

Hexavalent chromium, Cr(VI), is toxic to living systems. Widespread contamination of water and soil by Cr(VI) present a serious public health problem. Chromium-resistant bacteria can reduce and detoxify Cr(VI). Twelve bacteria resistant to high concentrations of Cr(VI) were isolated from soil enrichment cultures. Environmental parameters and kinetic parameters of Cr(VI) bioreduction by one monoculture isolate, identified by 16S rRNA gene sequence as Bacillus sp. PB2, were studied. The optimal temperature for growth and Cr(VI) reduction was 35 degrees C. The isolate grew luxuriantly and substantially reduced Cr(VI) at initial pH 7.5 to 9. Maximal Cr(VI) bioreduction occurred at initial pH 8.0. Substantial Cr(VI) bioreduction was observed in salt media, but removal efficiency was inversely related to salt concentration (1-9%). Michaelis-Menten hyperbolic equation and the Lineweaver-Burk double reciprocal plot were comparatively employed to determine the k (m) and V (max) of Cr(VI) bioreduction. A k (m) of 82.5 microg mL(-1) and V (max) of 7.78 microg mL(-1) h(-1) were calculated by nonlinear regression analysis of the hyperbola curve. Linear regression analysis of the double reciprocal plot revealed k (m) and V (max) of 80.9 microg mL(-1) and 10.6 microg mL(-1) h(-1), respectively. Time course studies displayed about 90% reduction of Cr(VI) at an initial concentration of 8,000 microg L(-1) in 8 h, with an estimated t (1/2) of 4 h. Data from time course analysis of the rate of Cr(VI) bioreduction fitted zero-order model, and the kinetic constant k was calculated to be 840 microg L(-1) h(-1). The monoculture isolate, Bacillus sp. PB2, strongly reduces Cr(VI) and could be used for bioremediation of Cr(VI)-contaminated aquatic and terrestrial environments.     

Apoptosis of lymphocytes induced by chromium(VI/V) is through ROS-mediated activation of Src-family kinases and caspase-3

Mechanistic insights into Cr(VI)-induced carcinogenicity and possible implication of Cr(V) species formed by the redox reactions of chromium-bearing species have attracted interest. We have previously demonstrated that when human peripheral blood lymphocytes are exposed to the Cr(V) complexes, viz., sodium bis(2-ethyl-2-hydroxybutyrato)oxochromate(V), Na[Cr(V)O(ehba)(2)] and sodium bis(2-hydroxy-2-methylbutyrato)oxochromate(V), Na[Cr(V)O(hmba)(2)], apoptosis and formation of reactive oxygen species (ROS) are observed. The molecular mechanisms involving cellular signaling pathways leading to apoptosis are addressed in the present study. Treatment of lymphocytes with Na[Cr(V)O(ehba)(2)] and K(2)Cr(2)O(7) leads to the activation of the Src-family protein tyrosine kinases namely, p56(lck), p59(fyn), and p56/53(lyn), which then activates caspase-3, both of which are under the partial influence of ROS. Inhibition of the Src-family tyrosine kinases activity by PP2 and of caspase-3 by Z-DEVD-FMK reverses apoptosis, thereby suggesting their importance. Antioxidants only partially reverse the apoptosis induced by Cr(VI/V), suggesting that pathways other than those induced by ROS cannot be ruled out. Although the complex, Na[Cr(V)O(ehba)(2)] is known to be relatively stable in aqueous solutions, previous studies have shown that the Cr(V) complex, Na[Cr(V)O(ehba)(2)] disproportionates to Cr(VI) and Cr(III) forms at pH 7.4 through complex mechanistic processes. Dynamics studies employing EPR data show that the Cr(V) state in Na[Cr(V)O(ehba)(2)] is relatively more stable in RPMI-1640 medium containing plasma. Formation of ROS during the reaction of redox partners with Na[Cr(V)O(ehba)(2)] is an early event and compares favorably in kinetic terms with the reported rate processes for disproportionation. This investigation presents evidence for the direct implication of Cr(V) in Cr(VI)-induced apoptosis of lymphocytes.     

Bacterial sensors based on Acidithiobacillus ferrooxidans Part II. Cr(VI) determination

The aerobic acidophilic bacterium Acidithiobacillus ferrooxidans oxidizes Fe(2+) and S(2)O(3)(2-) ions by consuming oxygen. An amperometric biosensor was designed including an oxygen probe as transducer and a recognition element immobilized by a suitable home-made membrane. This biosensor was used for the indirect amperometric determination of Cr(2)O(7)(2-) ions owing to methods based on a mediator (Fe(2+)) or titration. Using the mediator, the biosensor response versus Cr(2)O(7)(2-) was linear up to 0.4 mmol L(-1), with a response time of, respectively, 51 s (2 x 10(-5) mol L(-1) Cr(2)O(7)(2-)) and 61 s (6 x 10(-5) mol L(-1) Cr(2)O(7)(2-)). The method sensitivity was 816 microA L mol(-1). Response time and measurement sensitivity depended on membrane material and technique for biomass immobilization. For example, their values were 90 s-200 microA L mol(-1) when using a glass-felt membrane and 540 s-4.95 microA L mol(-1) with a carbon felt one to determine a concentration of 2 x 10(-5) mol L(-1) Cr(2)O(7)(2-). For the titration method, the biosensor is used to determine the equivalence point. The relative error of quantitative analysis was lower than 5%.     

Reduction of toxic hexavalent chromium by Ochrobactrum intermedium strain SDCr-5 stimulated by heavy metals

A Cr(VI) resistant bacterial strain SDCr-5, identified as Ochrobactrum intermedium on the basis of 16S rRNA gene sequencing, was tolerant to high concentrations of Cr(VI) up to 15 mg ml(-1) in acetate minimal medium. O. intermedium SDCr-5 reduced Cr(VI) under a wide range of concentrations from 100 to 1500 microg ml(-1) and reduction was optimum at 37 degrees C and pH 7. It reduced 200 and 721 microg ml(-1) Cr(VI) within 72 and 96 h, respectively. The rate of Cr(VI) reduction increased with concentration from 100 to 1500 microg ml(-1). The presence of heavy metal cations such as Cu(2+), Co(2+), Mn(2+) and Ni(2+) stimulated Cr(VI) reduction. Strain SDCr-5 might be useful for Cr(VI) detoxification under a wide range of environmental conditions.     

Sorption of Cr(VI), Cu(II) and Pb(II) by growing and non-growing cells of a bacterial consortium

This paper reports the sorption of three metallic ions, namely Cr(VI), Cu(II) and Pb(II) in aqueous solution by a consortium culture (CC) comprising an acclimatised mixed bacterial culture collected from point and non-point sources. Metal sorption capability of growing and non-growing cells at initial pH of between 3 and 8 in the 1-100mg/L concentration range were studied based on Q(max) and K(f) values of the Langmuir and linearised Freundlich isotherm models, respectively. Maximal metal loading was generally observed to be dependent on the initial pH. Growing cells displayed significant maximal loading (Q(max)) for Pb(II) (238.09 mg/g) and Cu(II) (178.87 mg/g) at pH 6 and at pH 7 for Cr(VI) (90.91 mg/g) compared to non-growing cells (p < 0.05). At the pH range of 6-8, growing cells showed higher loading capacity compared to non-growing cells i.e. 38-52% for Cr, 17-28% for Cu and 3-17% for Pb. At lower metal concentrations and at more acidic pH (3-4) however, non-growing cells had higher metal loading capacity than growing cells. The metal sorption capacity for both populations were as follows: Pb(II) > Cu(II) > Cr(VI).     

Characteristics of a novel Acinetobacter sp. and its kinetics in hexavalent chromium bioreduction

Cr-B2, a Gram-negative hexavalent chromium [Cr(VI)] reducing bacteria, was isolated from the aerator water of an activated sludge process in the wastewater treatment facility of a dye and pigment based chemical industry. Cr- B2 exhibited a resistance for 1,100 mg/l Cr(VI) and, similarly, resistance against other heavy metal ions such as Ni2+ (800 mg/l), Cu2+ (600 mg/l), Pb2+ (1,100 mg/l), Cd2+ (350 mg/l), Zn2+ (700 mg/l), and Fe3+ (1,000 mg/l), and against selected antibiotics. Cr-B2 was observed to efficiently reduce 200 mg/l Cr(VI) completely in both nutrient and LB media, and could convert Cr(VI) to Cr(III) aerobically. Cr(VI) reduction kinetics followed allosteric enzyme kinetics. The Km values were found to be 43.11 mg/l for nutrient media and 38.05 mg/l for LB media. Vmax values of 13.17 mg/l/h and 12.53 mg/l/h were obtained for nutrient media and LB media, respectively, and the cooperativity coefficients (n) were found to be 8.47 and 3.49, respectively, indicating positive cooperativity in both cases. SEM analysis showed the formation of wrinkles and depressions in the cells when exposed to an 800 mg/l Cr(VI) concentration. The organism was seen to exhibit pleomorphic behavior. Cr-B2 was identified on the basis of morphological, biochemical, and partial 16S rRNA gene sequencing chracterizations and found to be Acinetobacter sp.     

Isolation and Characterization of Chromium(VI)-Reducing Bacteria from Tannery Effluents

Two chromium-resistant bacteria (IFR-2 and IFR-3) capable of reducing/transforming Cr(VI) to Cr(III) were isolated from tannery effluents. Isolates IFR-2 and IFR-3 were identified as Staphylococcus aureus and Pediococcus pentosaceus respectively by 16S rRNA gene sequence analyses. Both isolates can grow well on 2,000 mg/l Cr(VI) (as K₂Cr₂O₇) in Luria-Bertani (LB) medium. Reduction of Cr(VI) was found to be growth-associated in both isolates and IFR-2 and IFR-3 reduced 20 mg/l Cr(VI) completely in 6 and 24 h respectively. The Cr(VI) reduction due to chromate reductase activity was detected in the culture supernatant and cell lysate but not at all in the cell extract supernatant of both isolates. Whole cells of IFR-2 and IFR-3 converted 24 and 30% of the initial Cr(VI) concentration (1 mg/l) in 45 min respectively at 37°C. NiCl₂ stimulated the growth of IFR-2 whereas HgCl₂ and CdCl₂ significantly inhibited the growth of both isolates. Optimum temperature and pH for growth of and Cr(VI) reduction by both isolates were found to be between 35 and 40°C and pH 7.0 to 8.0. The two bacterial isolates can be good candidates for detoxification of Cr(VI) in industrial effluents.     

Chromium Removal from Aqueous System and Industrial Wastewater by Agricultural Wastes

This study involved the development of formaldehyde-treated, deseeded sunflower head waste–based biosorbent (FSH) for the biosorption of Cr(VI) from aqueous solution and industrial wastewater. Batch-mode experiments were conducted to determine the kinetics, sorption isotherms, effect of pH, initial Cr(VI) concentration, biosorbent dose, and contact time. The results demonstrated that FSH can sequester Cr(VI) from the aqueous solution. The maximum sorption occurred at pH = 2.0, biosorbent dose = 4.0 g/L, concentration of 100 mg/L at 25°C at 180 rpm after 2 h contact time. The FSH had an adsorption capacity of 7.85 mg/g for Cr(VI) removal at pH 2.0. The rate of adsorption was rapid, and equilibrium was attained within 2 h. The equilibrium sorption data fitted the Langmuir isotherm model, which was further confirmed by the chi-square test.     

Chromate reduction by cell-free extract of Bacillus firmus KUCr1

Microbial enzymatic reduction of a toxic form of chromium [Cr(VI)] has been considered as an effective method for bioremediation of this metal. This study reports on the in vitro reduction of Cr(VI) using cell-free extracts from a Cr(VI) reducing Bacillus firmus KUCr1 strain. Chromium reductase was found to be constitutive and its activity was observed both in soluble cell fractions (S12 and S150 and membrane cell fraction (P150). The reductase activity of S12 fraction was found to be optimal at 40 microM Cr(VI) with enzyme concentration equivalent to 0.493 mg protein/ml. Enzyme activity was dependent on NADH or NADPH as electron donor; optimal temperature and pH for better enzyme activity were 70 degrees C and 5.6, respectively. The Km value of the reductase was 58.33 microM chromate having a V(max) of 11.42 microM/min/mg protein. The metabolic inhibitor like sodium azide inhibited reductase activity of membrane fraction of the cell-free extract. Metal ions like Cu2+, Co2+, Ni2+ and As3+ stimulated the enzyme but others, such as Ag+, Hg2+, Zn2+, Mn2+, Cd2+ and Pb2+, inhibited Cr(VI) reductase activity.     

Modeling hexavalent chromium removal in a Bacillus sp. fixed-film bioreactor

A one-dimensional diffusion-reaction model was developed to simulate Cr(VI) reduction in a Bacillus sp. pure culture biofilm reactor with glucose as a sole supplied carbon and energy source. Substrate utilization and Cr(VI) reduction in the biofilm was best represented by a system of (second-order) partial differential equations (PDEs). The PDE system was solved by the (fourth-order) Runge-Kutta method adjusted for mass transport resistance using the (second-order) Crank-Nicholson and Backward Euler finite difference methods. A heuristic procedure (genetic search algorithm) was used to find global optimum values of Cr(VI) reduction and substrate utilization rate kinetic parameters. The fixed-film bioreactor system yielded higher values of the maximum specific Cr(VI) reduction rate coefficient and Cr(VI) reduction capacity (kmc = 0.062 1/h, and Rc = 0.13 mg/mg, respectively) than previously determined in batch reactors (kmc = 0.022 1/h and Rc = 0.012 mg/mg). The model predicted effluent Cr(VI) concentration well with 98.9% confidence (sigmay2 = 2.37 mg2/L2, N = 119) and effluent glucose with 96.4 % confidence (sigmay(w)2 = 5402 mg2/L2, N = 121, w = 100) over a wide range of Cr(VI) loadings (10-498 mg Cr(VI)/L/d).     

Removal of hexavalent chromium in tannery wastewater by Bacillus cereus

Bacillus cereus was used to remove chromium (Cr(VI)) from medium containing tannery wastewater under different conditions. The maximum rate of Cr(VI) removal was attained at a temperature of 37?°C, pH of 7.0-9.0, and biomass of 20 g/L when the initial Cr(VI) concentration was less than 50?mg/L. Under the optimum conditions, the Cr(VI) in tannery wastewater was treated with each cellular component of B. cereus to detect its ability to reduce Cr(VI). The results showed that the removal rate of Cr(VI) for the cell-free extracts could reach 92.70%, which was close to that of the whole cells (96.85%), indicating that the Cr(VI) reductase generated by B.?cereus is primarily intracellular. Additionally, during continuous culture of the B. cereus, the strain showed good consecutive growth and removal ability. After treatment of 20?mg/L Cr(VI) for 48?h, the B. cereus was observed by SEM and TEM-EDX. SEM images showed that the B.?cereus used to treat Cr(VI) grew well and had a uniform cellular size. TEM-EDX analysis revealed large quantities of chromium in the B. cereus cells used to treat Cr(VI). Overall, the results presented herein demonstrate that B. cereus can be used as a new biomaterial to remove Cr(VI) from tannery wastewater.     

Removal of Chromium(VI) Ions from Synthetic Solutions by the Fungus Penicillium purpurogenum

The ability of Penicillium purpurogenum to bind high amounts of chromium(VI) from aqueous solutions is demonstrated. Cr(VI) adsorption capacity increases with time during the first four hours and then leveled off toward the equilibrium adsorption capacity. Biosorption of Cr(VI) ions reached equilibrium in four hours. Binding of Cr(VI) ions with Penicillium purpurogenum biomass was clearly pH dependent. Cr(VI) loading capacity increased with increasing pH. The adsorption of Cr(VI) ions reached a plateau value at a pH of approx. 6.0. The maximum capacity of adsorption of Cr(VI) ions onto the fungal biomass was 36.5 mg/g. Adsorption behavior of Cr(VI) ions can be approximately described with the Langmuir equation. When applying the Langmuir model, the maximum adsorption capacity (Q_max) and the Langmuir constant were found to be 40 mg/g and 3.9 × 10^–3 mg/L. Elution of Cr(VI) ions was performed by means of 0.5 M HCl. It was possible to use the biomass of Penicillium purpurogenum for six cycles for biosorption.     

Influence of nickel (II) and chromium (VI) on the laboratory scale rotating biological contactor

High concentration of heavy metals is toxic for most microorganisms and cause strict damage in wastewater treatment operations and often a physico-chemical pretreatment prior to biological treatment is considered necessary. However, in this study it has been shown that biological systems can adapt to Ni (II) and Cr (VI) when their concentration is below 10 and 20 mg/L, respectively. The aim of this study was to evaluate the effect of Ni (II) and Cr (VI) on the lab-scale rotating biological contactor process. It was found that, addition of Ni (II) up to 10 mg/L did not reduce the chemical oxygen demand removal efficiency and on the contrary concentrations below 10 mg/L improved the performance. The influent Ni (II) concentration of 1 mg/L was the concentration where the treatment efficiency produced a maximum COD removal of 86.5%. Moreover, Ni (II) concentration above 10 mg/L was relatively toxic to the system and produced lower treatment efficiencies than the baseline study without Ni (II). Turbidity and suspended solids removals were not stimulated to a great extent with nickel. Addition of Ni (II) did not seem to affect the pH of the system during treatment. The dissolved oxygen concentration did not drop below 4 mg/L at all concentrations of Ni (II) indicating aerobic conditions prevailed in the system. Experiments conducted with Cr (VI) revealed that addition of Cr (VI) up to 20 mg/L did not reduce the COD removal efficiency and on the contrary concentrations below 20 mg/L improved the performance. The influent Cr (VI) concentration of 1 mg/L was the concentration where the treatment efficiency produced a maximum COD removal of 88%. Turbidity and SS removals were more efficient at 5 mg/L Cr (VI) concentration, rather than 1 mg/L, which lead to the conclusion that 5 mg/L Cr (VI) concentration is the optimum concentration, in terms of COD, turbidity and SS removals. Similar with Ni (II) experiments, addition of Cr (VI) did not significantly affect the pH value of the effluent. The DO concentration remained above 5 mg/L.     

Cr(VI) removal from aqueous solution by thermophilic denitrifying bacterium <Emphasis Type="Italic">Chelatococcus daeguensis</Emphasis> TAD1 in the presence of single and multiple heavy metals

Cr(VI) pollution is increasing continuously as a result of ongoing industrialization. In this study, we investigated the thermophilic denitrifying bacterium Chelatococcus daeguensis TAD1, isolated from the biofilm of a biotrickling filter used in nitrogen oxides (NO_X) removal, with respect to its ability to remove Cr(VI) from an aqueous solution. TAD1 was capable of reducing Cr(VI) from an initial concentration of 10 mg/L to non-detectable levels over a pH range of 7–9 and at a temperature range of 30–50°C. TAD1 simultaneously removed both Cr(VI) and NO₃^?-N at 50°C, when the pH was 7 and the initial Cr(VI) concentration was 15 mg/L. The reduction of Cr(VI) to Cr(III) correlated with the growth metabolic activity of TAD1. The presence of other heavy metals (Cu, Zn, and Ni) inhibited the ability of TAD1 to remove Cr(VI). The metals each individually inhibited Cr(VI) removal, and the extent of inhibition increased in a cooperative manner in the presence of a combination of the metals. The addition of biodegradable cellulose acetate microspheres (an adsorption material) weakened the toxicity of the heavy metals; in their presence, the Cr(VI) removal efficiency returned to a high level. The feasibility and applicability of simultaneous nitrate removal and Cr(VI) reduction by strain TAD1 is promising, and may be an effective biological method for the clean-up of wastewater.     

Development of a new Cr(VI)-biosorbent from agricultural biowaste

Among useless but abundant agricultural biowastes such as banana skin, green tea waste, oak leaf, walnut shell, peanut shell and rice husk, in this study, banana skin was screened as the most efficient biomaterial to remove toxic Cr(VI) from aqueous solution. X-ray photoelectron spectroscopy (XPS) study revealed that the mechanism of Cr(VI) biosorption by banana skin was its complete reduction into Cr(III) in both aqueous and solid phases and partial binding of the reduced-Cr(III), in the range of pH 1.5-4 tested. One gram of banana skin could reduce 249.6 (+/-4.2)mg of Cr(VI) at initial pH 1.5. Namely, Cr(VI)-reducing capacity of banana skin was four times higher than that of a common chemical Cr(VI)-reductant, FeSO(4).7H(2)O. To diminish undesirable/serious organic leaching from the biomaterial and to enhance removal efficiency of total Cr, its powder was immobilized within Ca-alginate bead. The developed Cr(VI)-biosorbent could completely reduce toxic Cr(VI) to less toxic Cr(III) and could remove almost of the reduced-Cr(III) from aqueous phase. On the basis of removal mechanisms of Cr(VI) and total Cr by the Cr(VI)-biosorbent, a kinetic model was derived and could be successfully used to predict their removal behaviors in aqueous phase. In conclusion, our Cr(VI)-biosorbent must be a potent candidate to substitute for chemical reductants as well as adsorbents for treating Cr(VI)-bearing wastewaters.     

Removal of chromate anions from aqueous stream by a cationic surfactant-modified yeast

The removal of chromate anions (CrO(4)(2-)) from aqueous solution by a cationic surfactant-modified yeast was studied in a batch system. Cetyl trimethyl ammonium bromide (CTAB) was used for biomass modification; it substantially improved the biosorption efficiency. The influences of solution pH, CrO(4)(2-) anion concentrations and biomass concentration on the biosorption efficiency were investigated. The biosorption of chromate anions by modified yeast was strongly affected by pH. The optimum pH for biosorption of CrO(4)(2-) by modified yeast was 4.5-5.5. Zeta potential values of modified and unmodified yeast were determined at various pH values. Concentrations ranging from 5.2 to 208 mg/l Cr(VI) were tested and the biosorptive removal efficiency of the metal ions from aqueous solution was more than 99.5%. Freundlich and Langmuir isotherms were used to evaluate the data and the regression constants were determined.     

Reduction of hexavalent chromium by human cytochrome b5: generation of hydroxyl radical and superoxide

The reduction of hexavalent chromium, Cr(VI), can generate reactive Cr intermediates and various types of oxidative stress. The potential role of human microsomal enzymes in free radical generation was examined using reconstituted proteoliposomes (PLs) containing purified cytochrome b(5) and NADPH:P450 reductase. Under aerobic conditions, the PLs reduced Cr(VI) to Cr(V) which was confirmed by ESR using isotopically pure (53)Cr(VI). When 5-diethoxyphosphoryl-5-methyl-1-pyrroline-N-oxide (DEPMPO) was included as a spin trap, a very prominent signal for the hydroxyl radical (HO()) adduct was observed as well as a smaller signal for the superoxide (O(2)(-)) adduct. These adducts were observed even at very low Cr(VI) concentrations (10 muM). NADPH, Cr(VI), O(2), and the PLs were all required for significant HO() generation. Superoxide dismutase eliminated the O(2)(-) adduct and resulted in a 30% increase in the HO() adduct. Catalase largely diminished the HO() adduct signal, indicating its dependence on H(2)O(2). Some sources of catalase were found to have Cr(VI)-reducing contaminants which could confound results, but a source of catalase free of these contaminants was used for these studies. Exogenous H(2)O(2) was not needed, indicating that it was generated by the PLs. Adding exogenous H(2)O(2), however, did increase the amount of DEPMPO/HO() adduct. The inclusion of formate yielded the carbon dioxide radical adduct of DEPMPO, and experiments with dimethyl sulfoxide (DMSO) plus the spin trap alpha-phenyl-N-tert-butylnitrone (PBN) yielded the methoxy and methyl radical adducts of PBN, confirming the generation of HO(). Quantification of the various species over time was consistent with a stoichiometric excess of HO() relative to the net amount of Cr(VI) reduced. This also represents the first demonstration of a role for cytochrome b(5) in the generation of HO(). Overall, the simultaneous generation of Cr(V) and H(2)O(2) by the PLs and the resulting generation of HO() at low Cr(VI) concentrations could have important implications for Cr(VI) toxicity.