scholarly journals Microbial abilities to detoxify chromate by reduction

2001 ◽  
Author(s):  
◽  
Neroshini Maistry

Hexavalent chromium [Cr(VI)] or chromate, is a toxic, water-soluble contaminant present in many soils and industrial eflluents. As a result of contaminated discharges from industrial applications, and inappropriate wastedisposal practices, significant amounts of chromate have found their way into the environment. This poses a health risk to man as well as animals and plants due to the carcinogenicity, mutagenicity, and teratogenicity of chromate. In man, acute, high level exposures to Cr(VI) can result in ulceration of the skin, eyes, and mucous membranes. Exposure of plants to Cr(VI) can result in reduced biomass production, and in extreme cases, death. Upon reduction ofCr(VI) to trivalent chromium [Cr(III], the toxic effects are significantly decreased because of a decrease in the solubility and bioavailability of Cr(III). Traditionally, Cr(VI) has been recovered from aqueous systems using processes exploiting the differential solubility properties described above. The use of chromate reducing bacteria represents a potential mechanism for the development of an efficacious, cost effective alternative to traditional chemical/physical processes for Cr(VI) recovery from the environment. Therefore, the aim of this research was to isolate and identify chromate reducing bacteria from soil, and characterise the chromate reductase enzyme in order to determine the potential of bacteria to detoxify chromate by reduction. Bacteria from soils and wastewater were examined for chromate reducing potential and identified on the basis of biochemical tests and API 20E. Organisms were isolated by the spread plate technique. Species of Pseudomonas maltophilia, Bacillus subtilis, Acinetobacter calcoaceticus, and Cellumonas cellasea were capable of catalyzing the reduction ofCr(VI) to Cr(IlI) in batch experiments. Reduction capability as high as 99% by the isolates was detected from an initial Cr(VI) concentration of 150 mg.L' in batch cultures. Chromate reduction was determined by means of the diphenylcarbazide method and total chromium was measured by atomic absorption spectroscopy. Pseudomonas maltophilia was observed to be the most suited organism for the efficient detoxification ofCr(VI) due to its wide temperature and pH requirements, low substrate utilization, and tolerance to heavy metal ions of'Cu', Cd2+,Zn2+,and Ni2+which commonly appear in industrial eflluents along with Cr(VI). Reduction rate in a batch reactor for this organism was calculated to be 1.75 mg.g+h'. Comparison of the rates of chromate reduction by Cr(VI) grown cells and cells grown without chromate indicated that the chromate reductase activity is constitutive. Reductase activity was detected by means of the lysozyme-EDTA method in aerobically grown cells, with highest specific activity in the cytoplasmic fraction of the cell. The Cr(VI)-reductase was found to be NAD(p)H-dependent and yielded an activity of 3.24 ml.I.mg' of protein in the cytoplasmic fraction. Once optimization of the parameters in the batch reactor was achieved, cells of Ps. maltophilia was immobilized into polyacrylamide gel and packed in a column. Mass balance studies indicated that ca 147 mg.L' chromate passing through the column undergoes reduction with an initial Cr(VI) concentration of 150 mg.L' resulting in a Cr(VI) reduction efficiency of98%. An amount of 0.11 mg.L' remained in the cells, 0.11 mg.L' in the cell wash water, and 1.65 mg.L' was unaccounted for in the mass balance. Chromate reduction rate in the continuous-upflow reactor system was calculated to be 5.34 mg.g'l.h', which was 3-fold higher than that calculated for the batch reactor. Chromium-contaminated industrial eflluent obtained from Sheffield, Natal, and Saayman Danks Electroplaters was pumped into the continuous-upflow reactor containing immobilized cells of Ps. maltophilia to determine the industrial applicability of the reactor to treat chromate-containing effluents. Complete Cr(VI) reduction

DYNA ◽  
2019 ◽  
Vol 86 (210) ◽  
pp. 254-261 ◽  
Author(s):  
Diego Alonso Restrepo Molina ◽  
María Carolina Rodríguez Daza ◽  
Mario Evelio Arias Zabala

Sodium nitrate is an essential food additive widely used in cured meats. The increased unfavorable perceptions about chemicals in foods and their repercussions on health have positioned the nitrite as undesirable compound in meat products. Natural and organic ingredients have taken an important place within the consumer preferences. Here, obtaining nitrite from natural source was achieved using nitrate-reducing bacteria Staphylococcus carnosus and S. xylosus. The strains pre-incubation into the celery-based medium allowed the stimulation of nitrate reductase activity. The increased values of substrate, the oxygen condition and pH influenced the nitrite generation significantly (p<0.05). The reduction rate from nitrate to nitrite was 50.85%, with a value 320.2 mg L-1 nitrite for 4h. This method presents characteristics comparable to those traditionally applied in the generation of conventional sodium nitrite and lead to the food industry to take advantage of fermentation processes to supply needs in meats and supplementary food products.


2000 ◽  
Vol 66 (5) ◽  
pp. 1788-1795 ◽  
Author(s):  
C. H. Park ◽  
M. Keyhan ◽  
B. Wielinga ◽  
S. Fendorf ◽  
A. Matin

ABSTRACT Cr(VI) (chromate) is a widespread environmental contaminant. Bacterial chromate reductases can convert soluble and toxic chromate to the insoluble and less toxic Cr(III). Bioremediation can therefore be effective in removing chromate from the environment, especially if the bacterial propensity for such removal is enhanced by genetic and biochemical engineering. To clone the chromate reductase-encoding gene, we purified to homogeneity (>600-fold purification) and characterized a novel soluble chromate reductase from Pseudomonas putida, using ammonium sulfate precipitation (55 to 70%), anion-exchange chromatography (DEAE Sepharose CL-6B), chromatofocusing (Polybuffer exchanger 94), and gel filtration (Superose 12 HR 10/30). The enzyme activity was dependent on NADH or NADPH; the temperature and pH optima for chromate reduction were 80°C and 5, respectively; and theKm was 374 μM, with aV max of 1.72 μmol/min/mg of protein. Sulfate inhibited the enzyme activity noncompetitively. The reductase activity remained virtually unaltered after 30 min of exposure to 50°C; even exposure to higher temperatures did not immediately inactivate the enzyme. X-ray absorption near-edge-structure spectra showed quantitative conversion of chromate to Cr(III) during the enzyme reaction.


2020 ◽  
Vol 86 (22) ◽  
Author(s):  
Audrey G. O’Neill ◽  
Brett A. Beaupre ◽  
Yuanzhang Zheng ◽  
Dali Liu ◽  
Graham R. Moran

ABSTRACT Soil bacteria can detoxify Cr(VI) ions by reduction. Within the last 2 decades, numerous reports of chromate reductase enzymes have been published. These reports describe catalytic reduction of chromate ions by specific enzymes. These enzymes each have sequence similarity to known redox-active flavoproteins. We investigated the enzyme NfoR from Staphylococcus aureus, which was reported to be upregulated in chromate-rich soils and to have chromate reductase activity (H. Han, Z. Ling, T. Zhou, R. Xu, et al., Sci Rep 7:15481, 2017, https://doi.org/10.1038/s41598-017-15588-y). We show that NfoR has structural similarity to known flavin mononucleotide (FMN) reductases and reduces FMN as a substrate. NfoR binds FMN with a dissociation constant of 0.4 μM. The enzyme then binds NADPH with a dissociation constant of 140 μM and reduces the flavin at a rate of 1,350 s−1. Turnover of the enzyme is apparently limited by the rate of product release that occurs, with a net rate constant of 0.45 s−1. The rate of product release limits the rate of observed chromate reduction, so the net rate of chromate reduction by NfoR is orders of magnitude lower than when this process occurs in solution. We propose that NfoR is an FMN reductase and that the criterion required to define chromate reduction as enzymatic has not been met. That NfoR expression is increased in the presence of chromate suggests that the survival adaption was to increase the net rate of chromate reduction by facile, adventitious redox processes. IMPORTANCE Chromate is a toxic by-product of multiple industrial processes. Chromate reduction is an important biological activity that ameliorates Cr(VI) toxicity. Numerous researchers have identified chromate reductase activity by observing chromate reduction. However, all identified chromate reductase enzymes have flavin as a cofactor or use a flavin as a substrate. We show here that NfoR, an enzyme claimed to be a chromate reductase, is in fact an FMN reductase. In addition, we show that reduction of a flavin is a viable way to transfer electrons to chromate but that it is unlikely to be the native function of enzymes. We propose that upregulation of a redox-active flavoprotein is a viable means to detoxify chromate that relies on adventitious reduction that is not catalyzed.


2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Pablo M. Fernández ◽  
María M. Martorell ◽  
Julia I. Fariña ◽  
Lucia I. C. Figueroa

Resistance of the indigenous strainsP. jadiniiM9 andP. anomalaM10, to high Cr6+concentrations and their ability to reduce chromium in culture medium was studied. The isolates were able to tolerate chromium concentrations up to 104 μg mL−1. Growth and reduction of Cr6+were dependent on incubation temperature, agitation, Cr6+concentration, and pH. Thus, in both studied strains the chromium removal was increased at 30°C with agitation. The optimum pH was different, with values of pH 3.0 and pH 7.0 in the case ofP. anomalaM10 and pH 7.0 usingP. jadiniiM9. Chromate reduction occurred both in intact cells (grown in culture medium) as well as in cell-free extracts. Chromate reductase activity could be related to cytosolic or membrane-associated proteins. The presence of a chromate reductase activity points out a possible role of an enzyme in Cr6+reduction.


2003 ◽  
Vol 38 (3) ◽  
pp. 483-497 ◽  
Author(s):  
Susan A. Baldwin ◽  
Al Henry Hodaly

Abstract Sediment from a wetland receiving runoff from a coal mine waste dump in the Elk River Valley of southeast British Columbia was assessed for potential selenium uptake. Selenite [SeO32-, Se(IV)] was found to adsorb to the washed sediment at pH 7 to 8, whereas no selenate [SeO42-, Se(VI)] was adsorbed, in the concentration range of 8 to 225 μg L-1 Se as selenite or selenate. Sulfate- and selenate-reducing bacterial activity was detected in the sediment. In the presence of sulfate-reducing bacteria growth medium, Se as selenate was reduced from 619(±53) μg L-1 to 15(±0.7) μg L-1, and in the presence of selenate-reducing bacteria growth medium, Se as selenate was reduced from 364(±66) mg L-1 to 22(±10) mg L-1. Semi-continuous microcosms containing sediment overlaid with selenate (500 μg L-1 Se) and sulfate (0.9 g L-1) containing water were amended with plant debris from the site or nutrients (lactate and fertilizer). Potential selenate reduction rate (0.76 h-1) was highest in the unamended microcosms. Amendment with plant debris from the site had a negative effect on selenate reduction rate in the short term (after one hour) and a positive effect on Se removal in the long term (after one week). This study suggests that wetland sediments at the mine site may be important sinks for Se.


2020 ◽  
Author(s):  
Jhuma Biswas ◽  
AK Paul

Abstract The present study was aimed to characterize the chromate reducing ability of cells and cell-free extract (CFE) of Halomonas smyrnensis KS802 (GenBank Accession No. KU982965) and evaluate their effectiveness in tannery effluents. Viable cells of the strain reduced 200 µM Cr(VI) in basal medium for halophiles (MH) in 10 h and was inversely proportional to Cr(VI) concentrations. The rapid reduction by cells (10⁹ cells/mL) was achieved with 7.5% NaCl, at pH 7 and 37°C which increased with increasing cell density (10¹° cells/mL). While acetate, Cu³⁺, Fe³⁺, SO₄²⁻, and CO₃²⁻ were stimulating the reduction, the inhibitors retarded the process significantly. The NADH-dependent chromate reduction of the CFE was found to be constitutive with Km and Vmax values of 56.58 µM and 3.37 µM/min/mg protein respectively. The optimal reductase activity of the CFE was evident at 200 µM Cr(VI), 10% NaCl, pH 8.0 and at 45°C. A higher concentration of CFE and electron donors increased the enzyme activity but was impacted negatively by toxic metals and anions. Both the cells and CFE were capable of reducing Cr(VI) remarkably from tannery effluent. FTIR and XRD spectra of chromate reducing cells confirmed possible complexation of reduced Cr-species with functional groups on cell surface.


2001 ◽  
Vol 67 (3) ◽  
pp. 1076-1084 ◽  
Author(s):  
Jeff McLean ◽  
Terry J. Beveridge

ABSTRACT A pseudomonad (CRB5) isolated from a decommissioned wood preservation site reduced toxic chromate [Cr(VI)] to an insoluble Cr(III) precipitate under aerobic and anaerobic conditions. CRB5 tolerated up to 520 mg of Cr(VI) liter−1 and reduced chromate in the presence of copper and arsenate. Under anaerobic conditions it also reduced Co(III) and U(VI), partially internalizing each metal. Metal precipitates were also found on the surface of the outer membrane and (sometimes) on a capsule. The results showed that chromate reduction by CRB5 was mediated by a soluble enzyme that was largely contained in the cytoplasm but also found outside of the cells. The crude reductase activity in the soluble fraction showed aKm of 23 mg liter−1 (437 μM) and a V max of 0.98 mg of Cr h−1 mg of protein−1 (317 nmol min−1 mg of protein−1). Minor membrane-associated Cr(VI) reduction under anaerobiosis may account for anaerobic reduction of chromate under nongrowth conditions with an organic electron donor present. Chromate reduction under both aerobic and anaerobic conditions may be a detoxification strategy for the bacterium which could be exploited to bioremediate chromate-contaminated or other toxic heavy metal-contaminated environments.


2020 ◽  
Vol 367 (20) ◽  
Author(s):  
Yulia V Bertsova ◽  
Ilya P Oleynikov ◽  
Alexander V Bogachev

ABSTRACT The cytoplasmic fumarate reductase of Klebsiella pneumoniae (FRD) is a monomeric protein which contains three prosthetic groups: noncovalently bound FMN and FAD plus a covalently bound FMN. In the present work, NADH is revealed to be an inherent electron donor for this enzyme. We found that the fumarate reductase activity of FRD significantly exceeds its NADH dehydrogenase activity. During the catalysis of NADH:fumarate oxidoreductase reaction, FRD turnover is limited by a very low rate (∼10/s) of electron transfer between the noncovalently and covalently bound FMN moieties. Induction of FRD synthesis in K. pneumoniae cells was observed only under anaerobic conditions in the presence of fumarate or malate. Enzymes with the FRD-like domain architecture are widely distributed among various bacteria and apparently comprise a new type of water-soluble NADH:fumarate oxidoreductases.


1998 ◽  
Vol 37 (4-5) ◽  
pp. 599-603 ◽  
Author(s):  
Ryoko Yamamoto-Ikemoto ◽  
Saburo Matsui ◽  
Tomoaki Komori ◽  
Edja. Kofi. Bosque-Hamilton

The interactions between filamentous sulfur bacteria (FSB), sulfate reducing bacteria (SRB) and poly-P accumulating bacteria (PAB) in the activated sludge of a municipal plant operated under anaerobic-oxic conditions were examined in batch experiments using return sludge (RAS) and settled sewage. Phosphate release and sulfate reduction occurred simultaneously under anaerobic conditions. SRB were more sensitive to temperature changes than PAB. SRB played an important role in the decomposition of propionate to acetate. When the sulfate reduction rates were high, there was a tendency for the maximum release of phosphate also to be high. This was explained by the fact that PAB utilized the acetate produced by SRB. Sulfur oxidizing bacteria were sensitive to temperature change. When the sulfate reduction rate was high, the sulfide oxidizing rate was also high and filamentous bulking occurred. The results showed that sulfate reduction was a cause of filamentous bulking due to Type 021N that could utilize reduced sulfur.


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