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result(s) for
"Mercuric reductase"
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Characterization of Bacillus nealsonii strain KBH10 capable of reducing aqueous mercury in laboratory-scale reactor
2021
The environmental release of mercury is continuously increasing with high degree of mobility, transformation and amplified toxicity. Improving remediation strategies is becoming increasingly important to achieve more stringent environmental safety standards. This study develops a laboratory-scale reactor for bioremediation of aqueous mercury using a biofilm-producing bacterial strain, KBH10, isolated from mercury-polluted soil. The strain was found resistant to 80 mg/L of HgCl2 and identified as Bacillus nealsonii via 16S rRNA gene sequence analysis. The strain KBH10 was characterized for optimum growth parameters and its mercury biotransformation potential was validated through mercuric reductase assay. A packed-bed column bioreactor was designed for biofilm-mediated mercury removal from artificially contaminated water and residual mercury was estimated. Strain KBH10 could grow at a range of temperature (20–50 °C) and pH (6.0–9.0) with optimum temperature established at 30 °C and pH 7.0. The optimum mercuric reductase activity (77.8 ± 1.7 U/mg) was reported at 30 °C and was stable at a temperature range of 20–50 °C. The residual mercury analysis of artificially contaminated water indicated 60.6 ± 1.5% reduction in mercury content within 5 h of exposure. This regenerative process of biofilm-mediated mercury removal in a packed-bed column bioreactor can provide new insight into its potential use in mercury bioremediation.
Journal Article
Isolation and characterization of mercuric reductase by newly isolated halophilic bacterium, Bacillus firmus MN8
2017
The current study was aimed at isolating and identifying the halophilic and halotolerant bacteria which can produce mercuric reductase in Gavkhuni wetland in Iran. Moreover, tracking and sequencing merA gene and kinetic properties of mercuric reductase in the selected strain were performed in this study. Soil samples were taken from Gavkhuni wetland and cultured in nutrient agar medium with 5% NaCl. To examine the tolerance of purified colonies to mercury, agar dilution method was administered. Similarly, the phylogenetic analysis based on 16SrRNA gene sequencing was conducted. To investigate enzyme activity of kinetic parameters, a spectrophotometer was used to measure the NADPH oxidation decrease at 340 n.m. The results showed that among the 21 halophilic and halotolerant strains isolated from Gavkhuni wetland, 4 were resistant to mercuric chloride. A strain designated MN8 was selected for further studies because it showed the highest resistance to mercury. According to phylogenetic sequencing of 16S rRNA gene and phenotypic characteristics, the strain was categorized in the Bacillus genus and nearly related to Bacillus firmus. This strain had merA gene. The mercuric reductase showed Vmax and Km values of 0.106 U/mg and 24.051 µM, respectively. Evaluation of different concentrations of NaCl at 37°C and pH=7.5 in mercuric reductase enzyme activity indicated that the enzyme shows 50% activity in concentration of 1.5 M. Optimum pH and temperature of enzyme activity were 7.5 and 35 °C, respectively. The results suggested that MN8 strain could be a proper candidate for bioremediation of mercury-contaminated environments such as industrial wastewaters.
Journal Article
Mercury-Tolerant Ensifer medicae Strains Display High Mercuric Reductase Activity and a Protective Effect on Nitrogen Fixation in Medicago truncatula Nodules Under Mercury Stress
by
Tavakoli Zaniani, Parinaz
,
Balomenos, Dimitrios
,
García-Rodríguez, Diego
in
Abundance
,
Alfalfa
,
Bacteria
2021
Mercury (Hg) is extremely toxic for all living organisms. Hg-tolerant symbiotic rhizobia have the potential to increase legume tolerance, and to our knowledge, the mechanisms underlying Hg tolerance in rhizobia have not been investigated to date. Rhizobial strains of
Ensifer medicae
,
Rhizobium leguminosarum
bv.
trifolii
and
Bradyrhizobium canariense
previously isolated from severely Hg-contaminated soils showed different levels of Hg tolerance. The ability of the strains to reduce mercury Hg
2+
to Hg
0
, a volatile and less toxic form of mercury, was assessed using a Hg volatilization assay. In general, tolerant strains displayed high mercuric reductase activity, which appeared to be inducible in some strains when grown at a sub-lethal HgCl
2
concentration. A strong correlation between Hg tolerance and mercuric reductase activity was observed for
E. medicae
strains, whereas this was not the case for the
B. canariense
strains, suggesting that additional Hg tolerance mechanisms could be playing a role in
B. canariense
. Transcript abundance from
merA
, the gene that encodes mercuric reductase, was quantified in tolerant and sensitive
E. medicae
and
R. leguminosarum
strains. Tolerant strains presented higher
merA
expression than sensitive ones, and an increase in transcript abundance was observed for some strains when bacteria were grown in the presence of a sub-lethal HgCl
2
concentration. These results suggest a regulation of mercuric reductase in rhizobia. Expression of
merA
genes and mercuric reductase activity were confirmed in
Medicago truncatula
nodules formed by a sensitive or a tolerant
E. medicae
strain. Transcript accumulation in nodules formed by the tolerant strain increased when Hg stress was applied, while a significant decrease in expression occurred upon stress application in nodules formed by the Hg-sensitive strain. The effect of Hg stress on nitrogen fixation was evaluated, and in our experimental conditions, nitrogenase activity was not affected in nodules formed by the tolerant strain, while a significant decrease in activity was observed in nodules elicited by the Hg-sensitive bacteria. Our results suggest that the combination of tolerant legumes with tolerant rhizobia constitutes a potentially powerful tool in the bioremediation of Hg-contaminated soils.
Journal Article
Methylmercury demethylation and volatilization by animals expressing microbial enzymes
2025
Mercury is a highly toxic trace metal that readily biomagnifies in food webs where it is inaccessible to current bioremediation methods. Animals could potentially be engineered to detoxify mercury within their food webs to clean up impacted ecosystems. We demonstrate that invertebrate (
Drosophila melanogaster
) and vertebrate (
Danio rerio
) animal models can express organomercurial lyase (MerB) and mercuric reductase (MerA) from
Escherichia coli
to demethylate methylmercury and remove it from their biomass as volatile elemental mercury. The engineered animals accumulated less than half as much mercury relative to their wild-type counterparts, and a higher proportion of mercury in their tissue was in the form of less bioavailable inorganic mercury. Furthermore, the engineered animals could tolerate higher exposures to methylmercury compared to controls. These findings demonstrate the potential of using engineered animals for bioremediation and may be applied to reduce the burden of methylmercury in impacted ecosystems by disrupting its biomagnification or to treat contaminated organic waste streams.
Mercury is a highly toxic trace metal that readily biomagnifies in food webs where it is inaccessible to current bioremediation methods. Here the authors express organomercurial lyase and mercuric reductase from Escherichia coli in Drosophila melanogaster and Danio rerio to demethylate methylmercury and remove it from their biomass as volatile elemental mercury.
Journal Article
Characterization of a marine-isolated mercury-resistant Pseudomonas putida strain SP1 and its potential application in marine mercury reduction
2012
The Pseudomonas putida strain SP1 was isolated from marine environment and was found to be resistant to 280 μM HgCl^sub 2^. SP1 was also highly resistant to other metals, including CdCl^sub 2^, CoCl^sub 2^, CrCl^sub 3^, CuCl^sub 2^, PbCl^sub 2^, and ZnSO^sub 4^, and the antibiotics ampicillin (Ap), kanamycin (Kn), chloramphenicol (Cm), and tetracycline (Tc). mer operon, possessed by most mercury-resistant bacteria, and other diverse types of resistant determinants were all located on the bacterial chromosome. Cold vapor atomic absorption spectrometry and a volatilization test indicated that the isolated P. putida SP1 was able to volatilize almost 100% of the total mercury it was exposed to and could potentially be used for bioremediation in marine environments. The optimal pH for the growth of P. putida SP1 in the presence of HgCl^sub 2^ and the removal of HgCl^sub 2^ by P. putida SP1 was between 8.0 and 9.0, whereas the optimal pH for the expression of merA, the mercuric reductase enzyme in mer operon that reduces reactive Hg^sup 2+^ to volatile and relatively inert monoatomic Hg^sup 0^ vapor, was around 5.0. LD^sub 50^ of P. putida SP1 to flounder and turbot was 1.5×10^sup 9^ CFU. Biofilm developed by P. putida SP1 was 1- to 3-fold lower than biofilm developed by an aquatic pathogen Pseudomonas fluorescens TSS. The results of this study indicate that P. putida SP1 is a low virulence strain that can potentially be applied in the bioremediation of HgCl^sub 2^ contamination over a broad range of pH. [PUBLICATION ABSTRACT]
Journal Article
Characterization of Mercury-Resistant Rhizobacteria for Plant Growth Promotion: An In Vitro and In Silico Approach
by
Arslan, Sarwar
,
Amin Aatif
,
Muhammad, Naveed
in
Acetic acid
,
Biochemical tests
,
Bioinformatics
2021
In this study, a total 30 rhizobacterial isolates were screened out based on resistance against different concentrations of mercuric chloride (HgCl2), growth on nitrogen-free mannitol (NFM) and production of indole-3-acetic acid (IAA). The biochemical and plant growth promoting characterization of selected isolates was performed by different biochemical tests. Out of 30, six isolates, UM-3, AZ-5, UM-7, UM-11, UM-26, and UM-28 showed resistance at 30 µg/ml HgCl2, pronounced growth on NFM and high production of IAA as 18.6, 16.7, 16, 18.7, 14, and 16 µg/ml, respectively (P < 0.05). The 16S rDNA ribotyping and phylogenetic analysis of selected bacterial isolates were performed and characterized as Exiguobacterium sp. UM-3 (KJ736011), Bacillus thuringiensis AZ-5 (KJ675627), Bacillus subtilis UM-7 (KJ736013), Enterobacter cloacae UM-11 (KJ736014), Pseudomonas aeruginosa UM-26 (KJ736016), P. aeruginosa UM-28 (KJ736017) and Bacillus pumilus UM-16 (KJ736015) used as negative control. B. thuringiensis AZ-5 showed high resistance against 30 µg/ml of HgCl2 due to the presence of merB gene. The structural determination of MerB protein was carried out using bioinformatics tools, i.e., Protparam, Pfam, InterProScan, STRING, Jpred4, PSIPRED, I-TASSER, COACH server and ERRAT. These tools predicted the structural based functional homology of MerB protein (organomercuric lyase) in association with MerA (mercuric reductase) in bacterial Hg-detoxification system.
Journal Article
Plant colonizers of a mercury contaminated site: trace metals and associated rhizosphere bacteria
2024
Background and aims
Mercury (Hg) contamination poses severe human and environmental health risks. We aimed to evaluate the colonization of Hg-contaminated sites by native plants and the prokaryotic composition of rhizosphere soil communities of the dominant plant species.
Methods
A field study was conducted at a Hg-contaminated site in Romania. Metal concentrations in soil and plant samples were analyzed using portable X-ray fluorescence spectrometry. The prokaryotic composition of rhizosphere soil communities was determined through 16S rRNA amplicon sequencing and community functionality was predicted through PICRUSt2.
Results
Site-specific trace metal distribution across the site drove plant species distribution in the highly contaminated soil, with
Lotus tenuis
and
Diplotaxis muralis
associated with higher Hg concentrations. In addition, for the bacterial communities in the rhizosphere soil of
D. muralis
, there was no observable decrease in alpha diversity with increasing soil Hg levels. Notably, Actinomycetota had an average of 24% relative abundance in the rhizosphere communities that also tested positive for the presence of
merA
, whereas in the absence of
merA
the phylum’s relative abundance was approximately 2%.
merA
positive rhizosphere communities also displayed an inferred increase in ABC transporters.
Conclusions
The results suggest a dependence of species-wise plant survival on local trace metal levels in soil, as well as an intricate interplay of the latter with rhizosphere bacterial diversity. Knowledge of these interdependencies could have implications for phytoremediation stakeholders, as it may allow for the selection of plant species and appropriate soil microbial inoculates with elevated Hg tolerance.
Journal Article
Characterization of the Metabolically Modified Heavy Metal-Resistant Cupriavidus metallidurans Strain MSR33 Generated for Mercury Bioremediation
2011
Mercury-polluted environments are often contaminated with other heavy metals. Therefore, bacteria with resistance to several heavy metals may be useful for bioremediation. Cupriavidus metallidurans CH34 is a model heavy metal-resistant bacterium, but possesses a low resistance to mercury compounds.
To improve inorganic and organic mercury resistance of strain CH34, the IncP-1β plasmid pTP6 that provides novel merB, merG genes and additional other mer genes was introduced into the bacterium by biparental mating. The transconjugant Cupriavidus metallidurans strain MSR33 was genetically and biochemically characterized. Strain MSR33 maintained stably the plasmid pTP6 over 70 generations under non-selective conditions. The organomercurial lyase protein MerB and the mercuric reductase MerA of strain MSR33 were synthesized in presence of Hg(2+). The minimum inhibitory concentrations (mM) for strain MSR33 were: Hg(2+), 0.12 and CH(3)Hg(+), 0.08. The addition of Hg(2+) (0.04 mM) at exponential phase had not an effect on the growth rate of strain MSR33. In contrast, after Hg(2+) addition at exponential phase the parental strain CH34 showed an immediate cessation of cell growth. During exposure to Hg(2+) no effects in the morphology of MSR33 cells were observed, whereas CH34 cells exposed to Hg(2+) showed a fuzzy outer membrane. Bioremediation with strain MSR33 of two mercury-contaminated aqueous solutions was evaluated. Hg(2+) (0.10 and 0.15 mM) was completely volatilized by strain MSR33 from the polluted waters in presence of thioglycolate (5 mM) after 2 h.
A broad-spectrum mercury-resistant strain MSR33 was generated by incorporation of plasmid pTP6 that was directly isolated from the environment into C. metallidurans CH34. Strain MSR33 is capable to remove mercury from polluted waters. This is the first study to use an IncP-1β plasmid directly isolated from the environment, to generate a novel and stable bacterial strain useful for mercury bioremediation.
Journal Article
An implication of biotransformation in detoxification of mercury contamination by Morganella sp. strain IITISM23
2021
The contamination of soil by heavy metals such as Hg is growing immensely nowadays. The drawbacks of physicochemical methods in the decontamination of polluted soils resulted in the search for an eco-friendly and cost-effective means in this regard. In this study, a potential Hg-resistant bacterial (IITISM23) strain was investigated for their removal potential of Hg, isolated from Hg-contaminated soil. IITISM23 strain was identified as
Morganella
sp. (MT062474.1) as it showed 99% similarity to genus
Morganella
of
Gammaproteobacteria
based on 16S rRNA gene sequencing. The toxicity experiment confirmed that the strain showed high resistance toward Hg. In low nutrient medium, EC
50
(effective concentration) values were 6.8 ppm and minimum effective concentration (MIC) was 7.3 ppm, and in a nutrient-rich medium, EC
50
value was 32.29 ppm and MIC value was 34.92 ppm, respectively. In in vitro conditions, IITISM23 showed the removal efficiency (81%) of Hg (II) by the volatilization method in Luria-Bertani (LB) broth. The changes in surface morphology of bacteria upon the supplementation of Hg (II) in broth media were determined by SEM-EDX studies, while the changes in functional groups were studied by FT-IR spectroscopy. The mercury reductase activity was determined by a crude extract of the bacterial strain. The optimal pH and temperature for maximum enzyme activity were 8 and 30
o
C, with Km of 3.5 μmol/l and Vmax of 0.88 μmol/min, respectively. Also, strain IITISM23 showed resistance toward various antibiotics and other heavy metals like cadmium, lead, arsenic, and zinc. Hence, the application of microbes can be an effective measure in the decontamination of Hg from polluted soils.
Journal Article
A Rhizosphere-Associated Symbiont, Photobacterium spp. Strain MELD1, and Its Targeted Synergistic Activity for Phytoprotection against Mercury
by
Mathew, Gincy Marina
,
Gicana, Ronnie Gicaraya
,
Chien, Mei-Chieh
in
Acetic acid
,
Agricultural production
,
Aquatic plants
2015
Though heavy metal such as mercury is toxic to plants and microorganisms, the synergistic activity between them may offer benefit for surviving. In this study, a mercury-reducing bacterium, Photobacterium spp. strain MELD1, with an MIC of 33 mg x kg(-1) mercury was isolated from a severely mercury and dioxin contaminated rhizosphere soil of reed (Phragmites australis). While the whole genome sequencing of MELD1 confirmed the presence of a mer operon, the mercury reductase MerA gene showed 99% sequence identity to Vibrio shilloni AK1 and implicates its route resulted from the event of horizontal gene transfer. The efficiency of MELD1 to vaporize mercury (25 mg x kg(-1), 24 h) and its tolerance to toxic metals and xenobiotics such as lead, cadmium, pentachlorophenol, pentachloroethylene, 3-chlorobenzoic acid, 2,3,7,8-tetrachlorodibenzo-p-dioxin and 1,2,3,7,8,9-hexachlorodibenzo-p-dioxin is promising. Combination of a long yard bean (Vigna unguiculata ssp. Sesquipedalis) and strain MELD1 proved beneficial in the phytoprotection of mercury in vivo. The effect of mercury (Hg) on growth, distribution and tolerance was examined in root, shoot, leaves and pod of yard long bean with and without the inoculation of strain MELD1. The model plant inoculated with MELD1 had significant increases in biomass, root length, seed number, and increased mercury uptake limited to roots. Biolog plate assay were used to assess the sole-carbon source utilization pattern of the isolate and Indole-3-acetic acid (IAA) productivity was analyzed to examine if the strain could contribute to plant growth. The results of this study suggest that, as a rhizosphere-associated symbiont, the synergistic activity between the plant and MELD1 can improve the efficiency for phytoprotection, phytostabilization and phytoremediation of mercury.
Journal Article