Mercuric Reductase Gene Research Articles

Impacts of Selenium Supplementation on Soil Mercury Speciation, Soil Properties and Mercury-Resistant Microorganisms and Resistant Genes

Soil mercury (Hg) contamination is a serious threat to local ecology and public health. Exogenous selenium (Se) supplementation can effectively reduce the toxicity of Hg. However, the mechanisms affecting the changes in soil Hg speciation, soil properties and the microbial Hg-resistant system during the Se–Hg interaction after exogenous Se supplementation are not clear. Therefore, in this study, soil culture experiments were conducted to analyze the effects of different Se additions on the transformation of Hg speciation, soil properties and Hg-resistant microorganisms and resistant genes (mer operon). The results indicated that Se supplementation facilitated the transformation of soil Hg from bioavailable (exchangeable and carbonate-bound) to stable forms (organic material-bound and residual), significantly reducing Hg bioavailability. Se supplementation notably decreased the electrical conductivity of Hg-contaminated soil, but had no significant effect on the soil pH, organic matter content, cation exchange capacity or alkaline phosphatase and catalase activities. The maximum activity levels of soil sucrase and urease were observed when 1 mg kg−1 Se was added. Se significantly inhibited soil peroxidase and ascorbate oxidase activities, thereby alleviating the oxidative stress in the soil system caused by Hg. Additionally, Se significantly activated the Hg-resistant system in soil microorganisms by either decreasing or increasing the regulatory genes merD and merR, and it significantly upregulated the cytoplasmic protein gene merP and the membrane protein genes merC, merF and merT. This further increased the abundance of the organomercury lyase gene merB and the mercuric reductase gene merA, promoting the conversion of Hg species to Hg⁰. Furthermore, the abundance of mer operon-containing microorganisms, such as Thiobacillus ferrooxidants, Pseudomonas, Streptomyces and Cryptococcus, significantly increased with Se addition, explaining the role of soil microorganisms in mitigating soil Hg stress via Se supplementation.

Labile carbon inputs boost microbial contribution to legacy mercury reduction and emissions from industry-polluted soils

Soils is a crucial reservoir influencing mercury (Hg) emissions and soil-air exchange dynamics, partially modulated by microbial reducers aiding Hg reduction. Yet, the extent to which microbial engagements contribute to soil Hg volatilization remains largely unknown. Here, we characterized Hg-reducing bacterial communities in natural and anthropogenically perturbed soil environments and quantified their contribution to soil Hg(0) volatilization. Our results revealed distinct Hg-reducing bacterial compositions alongside elevated mercuric reductase (merA) gene abundance and diversity in soils adjacent to chemical factories compared to less-impacted ecosystems. Notably, solely industry-impacted soils exhibited increased merA gene abundance along Hg gradients, indicating microbial adaption to Hg selective pressure through quantitative changes in Hg reductase and genetic diversity. Microcosm studies demonstrated that glucose inputs boosted microbial involvement and induced 2–8 fold increments in cumulative Hg(0) volatilization in industry-impacted soils. Microbially-mediated Hg reduction contributed to 41.6% of soil Hg(0) volatilization in industry-impacted soils under 25% water-holding capacity and glucose input conditions over a 21-day incubation period. Alcaligenaceae, Moraxellaceae, Nitrosomonadaceae and Shewanellaceae were identified as potential contributors to Hg(0) volatilization in the soil. Collectively, our study provides novel insights into microbially-mediated Hg reduction and soil-air exchange processes, with important implications for risk assessment and management of industrial Hg-contaminated soils.

Unraveling the potential of bacteria isolated from the equatorial region of Indian Ocean in mercury detoxification

The marine environment is most vital and flexible with continual variations in salinity, temperature, and pressure. As a result, bacteria living in such an environment maintain the adaption mechanisms that are inherent in unstable environmental conditions. The harboring of metal-resistant genes in marine bacteria contributes to their effectiveness in metal remediation relative to their terrestrial counterparts. A total of four mercury-resistant bacteria (MRB) i.e. NIOT-EQR_J7 (Alcanivorax xenomutans); NIOT-EQR_J248 and NIOT-EQR_J251 (Halomonas sp.); and NIOT-EQR_J258 (Marinobacter hydrocarbonoclasticus) were isolated from the equatorial region of the Indian Ocean (ERIO) and identified by analyzing the 16S rDNA sequence. The MRBs can reduce up to 70% of Hg(II). The mercuric reductase (merA) gene was amplified and the mercury (Hg) volatilization was confirmed by the X-ray film method. The outcomes obtained from ICP-MS validated that the Halomonas sp. NIOT-EQR_J251 was more proficient in removing the Hg from culture media than other isolates. Fourier transform infrared (FT-IR) spectroscopy results revealed alteration in several functional groups attributing to the Hg tolerance and reduction. The Gas Chromatography-Mass Spectrometry (GC-MS) analysis confirmed that strain Halomonas sp. (NIOT-EQR_J248 and NIOT-EQR_J251) released Isooctyl thioglycolate (IOTG) compound under mercury stress. The molecular docking results suggested that IOTG can efficiently bind with the glutathione S-transferase (GST) enzyme. A pathway has been hypothesized based on the GC-MS metabolic profile and molecular docking results, suggesting that the compound IOTG may mediate mercuric reduction via merA-GST related detoxification pathway.

Mercuric reductase gene (merA) activity in a mercury tolerant sulphate reducing bacterium isolated from the Kongsfjorden, Arctic

The bacterial reduction of mercury is one of the transformation processes where ionic form (Hg2+) converts into elemental form (Hg0). The biological reduction of mercury is facilitated by the mercuric reductase (merA) gene. The significance of understanding the role of microbial mercury reduction in the Arctic during the changing climate conditions is high as it acts as the sink as well as a source of mercury. The present study was carried out to assess the prevalence of mercury tolerant bacteria and to know the distribution of merA in the Arctic bacterial communities. Citrobacter freundii strain MM7, a psychrotrophic, gram-negative, facultative anaerobic, non-traditional sulphate-reducing bacterium, isolated from the sediments of Kongsfjorden showed tolerance to mercury up to 10.0 mg/L in vitro. Mercury removal experiment confirmed the removal of ∼80% of mercury mediated by plasmid-borne merA gene. Studies using Fourier transform infrared (FTIR) showed the metabolic changes incited by mercury, and the spectroscopic differences reflected that the functional groups like alkynes, aromatic phosphates, organic phosphates, methyl, methylene, and amide groups present in the bacterial surface owe a significant part in Hg biosorption and removal. The present study provides insights on the mercury resistance in an Arctic bacterial strain and suggests the potential application of Citrobacter freundii strain MM7 in the bioremediation of mercury from the natural environment.

Tolerance of the forest soil microbiome to increasing mercury concentrations

Mercury (Hg) occurs naturally in soil and can reach high concentrations in the environment due to anthropogenic activities; however, little is known about its impact on the soil microbiome. Here, we tested the impact of different concentrations of Hg on soil bacterial and fungal communities by carrying out microcosm experiments with seven different forest soils. The highest concentration of Hg (32 μg Hg g−1 dry soil) caused severe diversity loss and shifts in the bacterial and fungal community structures and composition. Bioavailable Hg was reduced in soils with the highest proportion of clay, but the impact of Hg on microbial community structures was still evident in these soils. Lower concentrations of Hg (≤3.2 μg Hg g−1 dry soil) had only a limited effect on the soil microbiome. Fungal communities were generally less affected than the bacterial communities. The bacterial Hg-detoxification capacity, as assessed by mercuric reductase gene abundance, was reduced in soils with the lowest amount bioavailable Hg. We found a wide range of Hg-responsive taxa in soils spiked with high amounts of Hg, although they were generally not specific to any soil type or taxonomic group. Overall, our data show that the impacts of Hg on the soil microbiome and its detoxification responses depend on soil characteristics.

Mercury remediation potential of a mercury resistant strain Sphingopyxis sp. SE2 isolated from contaminated soil

A mercury resistant bacterial strain SE2 was isolated from contaminated soil. The 16s rRNA gene sequencing confirms the strain as Sphingopyxis belongs to the Sphingomonadaceae family of the α-Proteobacteria group. The isolate showed high resistance to mercury with estimated concentrations of Hg that caused 50% reduction in growth (EC50) of 5.97 and 6.22mg/L and minimum inhibitory concentrations (MICs) of 32.19 and 34.95mg/L in minimal and rich media, respectively. The qualitative detection of volatilized mercury and the presence of mercuric reductase enzyme proved that the strain SE2 can potentially remediate mercury. ICP-QQQ-MS analysis of the remaining mercury in experimental broths indicated that a maximum of 44% mercury was volatilized within 6hr by live SE2 culture. Furthermore a small quantity (23%) of mercury was accumulated in live cell pellets. While no volatilization was caused by dead cells, sorption of mercury was confirmed. The mercuric reductase gene merA was amplified and sequenced. Homology was observed among the amino acid sequences of mercuric reductase enzyme of different organisms from α-Proteobacteria and ascomycota groups.

Mercury resistance and volatilization by Pseudoxanthomonas sp. SE1 isolated from soil

Abstract A mercury resistant bacterial strain SE1 isolated from contaminated soil was identified as Pseudoxanthomonas based on 16s rRNA sequencing. The Hg resistance was examined in both nutrient-rich media as well as low nutrient media and expressed as EC50 and MIC values. Estimated EC50 and MIC values in nutrient-rich media and low nutrient media had the following respective recordings — 22.6 mg L−1; 23.1 mg L−1 and 1.4 mg L−1 and 1.7 mg L−1. The isolate was able to volatilize inorganic mercury demonstrated by a modified photographic film experiment and subsequently revealed its ability to remove mercury from the solution. The ICP-QQQ-MS analysis of SE1 inoculated solution showed almost 60% of 1.5 mg L−1 mercury was volatilized in 6 h and almost 40% were accumulated in cell pellets. The mercuric reductase gene merA was identified in the genome of isolate SE1 and sequenced. The deduced amino acid sequence of merA gene indicated a sequence homology with different organisms from the alpha proteobacteria group and eukaryotic fungi. merA encoded enzyme mercuric reductase activity was evident in the crude protein of the isolate. The isolate’s ability to resist Hg, it’s Hg volatilization potential and the presence of merA gene and mercuric reductase enzyme demonstrates the potential application of this strain in mercury bioremediation.

Methylmercury degradation by Pseudomonas putida V1

Environmental contamination of mercury (Hg) has caused public health concerns with focuses on the neurotoxic substance methylmercury, due to its bioaccumulation and biomagnification in food chains. The goals of the present study were to examine: (i) the transformation of methylmercury, thimerosal, phenylmercuric acetate and mercuric chloride by cultures of Pseudomonas putida V1, (ii) the presence of the genes merA and merB in P. putida V1, and (iii) the degradation pathways of methylmercury by P. putida V1. Strain V1 cultures readily degraded methylmercury, thimerosal, phenylmercury acetate, and reduced mercuric chloride into gaseous Hg(0). However, the Hg transformation in LB broth by P. putida V1 was influenced by the type of Hg compounds. The merA gene was detected in P. putida V1, on the other hand, the merB gene was not detected. The sequencing of this gene, showed high similarity (100%) to the mercuric reductase gene of other Pseudomonas spp. Furthermore, tests using radioactive 14C-methylmercury indicated an uncommon release of 14CO2 concomitant with the production of Hg0. The results of the present work suggest that P. putida V1 has the potential to remove methylmercury from contaminated sites. More studies are warranted to determine the mechanism of removal of methylmercury by P. putida V1.

Bioremediation potential of a highly mercury resistant bacterial strain Sphingobium SA2 isolated from contaminated soil

A mercury resistant bacterial strain, SA2, was isolated from soil contaminated with mercury. The 16S rRNA gene sequence of this isolate showed 99% sequence similarity to the genera Sphingobium and Sphingomonas of α-proteobacteria group. However, the isolate formed a distinct phyletic line with the genus Sphingobium suggesting the strain belongs to Sphingobium sp. Toxicity studies indicated resistance to high levels of mercury with estimated EC50 values 4.5 mg L−1 and 44.15 mg L−1 and MIC values 5.1 mg L−1 and 48.48 mg L−1 in minimal and rich media, respectively. The strain SA2 was able to volatilize mercury by producing mercuric reductase enzyme which makes it potential candidate for remediating mercury. ICP-QQQ-MS analysis of Hg supplemented culture solutions confirmed that almost 79% mercury in the culture suspension was volatilized in 6 h. A very small amount of mercury was observed to accumulate in cell pellets which was also evident according to ESEM-EDX analysis. The mercuric reductase gene merA was amplified and sequenced. The deduced amino acid sequence demonstrated sequence homology with α-proteobacteria and Ascomycota group.

Microbial DNA records historical delivery of anthropogenic mercury.

Mercury (Hg) is an anthropogenic pollutant that is toxic to wildlife and humans, but the response of remote ecosystems to globally distributed Hg is elusive. Here, we use DNA extracted from a dated sediment core to infer the response of microbes to historical Hg delivery. We observe a significant association between the mercuric reductase gene (merA) phylogeny and the timing of Hg deposition. Using relaxed molecular clock models, we show a significant increase in the scaled effective population size of the merA gene beginning ~200 years ago, coinciding with the Industrial Revolution and a coincident strong signal for positive selection acting on residues in the terminal region of the mercuric reductase. This rapid evolutionary response of microbes to changes in the delivery of anthropogenic Hg indicates that microbial genomes record ecosystem response to pollutant deposition in remote regions.

Mercuric reductase genes (merA) and mercury resistance plasmids in High Arctic snow, freshwater and sea-ice brine

Bacterial reduction in Hg(2+) to Hg(0) , mediated by the mercuric reductase (MerA), is important in the biogeochemical cycling of Hg in temperate environments. Little is known about the occurrence and diversity of merA in the Arctic. Seven merA determinants were identified among bacterial isolates from High Arctic snow, freshwater and sea-ice brine. Three determinants in Bacteriodetes, Firmicutes and Actinobacteria showed < 92% (amino acid) sequence similarity to known merA, while one merA homologue in Alphaproteobacteria and 3 homologues from Betaproteobacteria and Gammaproteobacteria were > 99% similar to known merA's. Phylogenetic analysis showed the Bacteroidetes merA to be part of an early lineage in the mer phylogeny, whereas the Betaproteobacteria and Gammaproteobacteria merA appeared to have evolved recently. Several isolates, in which merA was not detected, were able to reduce Hg(2+) , suggesting presence of unidentified merA genes. About 25% of the isolates contained plasmids, two of which encoded mer operons. One plasmid was a broad host-range IncP-α plasmid. No known incompatibility group could be assigned to the others. The presence of conjugative plasmids, and an incongruent distribution of merA within the taxonomic groups, suggests horizontal transfer of merA as a likely mechanism for High Arctic microbial communities to adapt to changing mercury concentration.

Isolation and molecular identification of mercury resistant bacteria and detection of Escherichia coli mercuric reductase gene from wastewater of Khowr-e-Musa, Iran

Mercuric compounds are extending over large natural environment as a result of industrial pollution. Resistance to these compounds have been found in a wide range of bacterial species isolated from various environment. The aim of this study was to investigate of mercury resistant bacteria on the seashore wastewater of Khowr-e-Musa in Mahshahr area, in the south west of Iran, which one of the most important petrochemical chlor-alkali unit is located there. For this purpose, water samples were taken from wastewater of three stations. Amount of total mercury in the samples was measured using cold vapor atomic absorption spectrophotometery. Two approaches namely, conventional biochemical test and modern molecular approaches were used for identification. Mercury toxicity was measured via minimal inhibitory concentration method. Bacillus cereus, E.coli and Staphylococcus aureus were isolated and identified based on 16S rRNA gene homology, and resistance to mercuric chloride was at 400, 450 and 75 ppm, respectively. The location of E.coli mer operon was determined by plasmid curing. E.coli showed the presence of a plasmid DNA which is carries mer operon, and 1695 bp of merA gene was amplified by PCR method. The results exhibited that isolated bacteria in present study were resistant and could grow on high concentration of mercury. * Corresponding Author: Farshid Kafilzadeh  kafilzadeh@jia.ac.ir

The effect of a continuous mercury stress on mercury reducing community of some characterized bacterial strains

Active resistance to the mercuric ion is widely distributed in environmental microbes and results from the action of mercuric reductase. Five mercury resistant bacteria:Escherichia coli Z1, Escherichia coli Z3, Pseudomonas putida Z2, Serratia marcescensZ4 and Xanthomonas sp. Z5 were isolated and identified from sludge sample. The presence of mercury resistance determinants was screened by PCR using merA-specific primers. Based on the analysis of merA amplicons, high similarity was recorded between the merA region of the strains P. putida Z2 and Xanthomonas sp. Z5 with those of Tn5053; while the merA of E. coli Z1 was analogous to those of Tn21. In case of the bacterial strains E. coli Z3 and S. marcescens Z4 a great matching was obtained between their merA and those of Tn5036. The effect of mercury stress upon the structure of mercury reducing biofilm at the species level and the type of mercury resistance determinants was studied in a continuous bioreactor. Community analysis suggested that the bacterial strain E. coli Z3 containing Tn5036-like determinant is the well adapted strain that tolerated elevated levels of mercury whereas the other strains showed a less fitness under these extreme conditions. Key words: Mercury resistant bacteria, mercuric reductase gene, PCR-RFLP, mercury stress.

Effects of long‐term fertilization on the diversity of bacterial mercuric reductase gene in a Chinese upland soil

Soil mercury (Hg) pollution has received considerable attention due to its neurotoxin effects and its potential risk to food safety. The microbial transformation of Hg plays a key role in reducing Hg toxicity by the mercuric reductase (MerA) conferred by genes arranged in the mer operon. This study investigated the effects of long-term fertilization on the diversity of bacterial mercuric reductase gene (merA), which specify the reduction of ionic Hg²⁺ to the volatile elemental form Hg⁰, in an agricultural soil with relatively high Hg content. The soil samples were collected from different treatments, including control without fertilizer (CK), fertilizer nitrogen (N), combined fertilizers (NPK) of N, phosphorus (P) and potassium (K), and NPK plus organic manure (NPK + OM). The merA gene diversity patterns were analyzed based on the merA clone libraries and sequencing measurements. Results showed that the merA gene diversity was influenced by soil variables depending on the fertilization practices. In particular, NH₄⁺ and NO₃⁻ contents had strong effect on the merA gene diversity pattern both in the N and NPK treatments, whereas the merA gene diversity pattern in NPK + OM treatment was distinctly influenced by the contents of organic matter, available P and K. These results suggested that long-term fertilization had significant influences on merA gene diversity, which could be helpful to understand the Hg reduction process and potentially serve microbial remediation of Hg contaminated soil.

Response to mercury (II) ions in Methylococcus capsulatus (Bath)

The mercury (II) ion is toxic and is usually detoxified in Bacteria by reduction to elemental mercury, which is less toxic. This is catalysed by an NAD(P)H-dependent mercuric reductase (EC 1.16.1.1). Here, we present strong evidence that Methylococcus capsulatus (Bath) - a methanotrophic member of the Gammaproteobacteria - uses this enzyme to detoxify mercury. In radiorespirometry studies, it was found that cells exposed to mercury dissimilated 100% of [(14) C]-methane provided to generate reducing equivalents to fuel mercury (II) reduction, rather than the mix of assimilation and dissimilation found in control incubations. The detoxification system is constitutively expressed with a specific activity of 352 (±18) nmol NADH oxidized min(-1) (mg protein)(-1) . Putative mercuric reductase genes were predicted in the M.capsulatus (Bath) genome and found in mRNA microarray studies. The MerA-derived polypeptide showed high identity (>80%) with MerA sequences from the Betaproteobacteria.

Molecular Cloning and Expression of Bacterial Mercuric Reductase Gene

In order to characterize the bacterial mercuric reductase (merA) gene, mercury resistant (Hgr) Escherichia coli strains have been isolated from various mercury contaminated sites of India. Their minimum inhibitory concentration (MIC) for Hg and zone of inhibition for different antibiotics were measured, and finally mer operon was localized by transforming isolated E. coli plasmid into mercury sensitive (Hgs) host E. coli DH5a cells. Oligonucleotide primers were designed by comparing the known reported sequences of merA from Gram-negative bacterium (E. coli plasmid R100) and 1695 bp full length merA gene was amplified by PCR. A 1.695-kb DNA fragment of merA was inserted into isopropyl-
-D-thiogalactopyranoside (IPTG) inducible bacterial expression vector pQE-30U/A. E. coli DH5 strains harboring the merA constructs showed higher mercury reductase enzyme (MerA) activity and expressed significantly more MerA than the control strains under aerobic conditions. The purified merA gene over expressed in the specific host E. coli BL21(DE3)Plys cells. Finally, expressed MerA protein was purified by Immobilized Metal-chelate Affinity Chromatography (IMAC) by using Ni- NTA column; and ~66.2 kDa bacterial MerA protein was detected after resolving on 10% sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS PAGE).

A conservative region of the mercuric reductase gene (merA) as a molecular marker of bacterial mercury resistance

The most common bacterial mercury resistance mechanism is based on the reduction of Hg(II) to Hg(0), which is dependent of the mercuric reductase enzyme (MerA) activity. The use of a 431 bp fragment of a conservative region of the mercuric reductase (merA) gene was applied as a molecular marker of this mechanism, allowing the identification of mercury resistant bacterial strains.

Mercury Speciation, Reactivity, and Bioavailability in a Highly Contaminated Estuary, Berry’s Creek, New Jersey Meadowlands

Speciation and reactivity of mercury were examined in Berry's Creek estuary downstream of a highly mercury-contaminated U.S. EPA Superfund site during the summers of 2002 and 2003. Surface water samples from Berry's Creek estuary, its confluence with the Hackensack River, and upstream of that confluence were analyzed for total (THg), particulate (PHgT), and dissolved (DHg) mercury, total and particulate monomethylmercury (MeHg), dissolved gaseous mercury (DGM), and bacterial merA gene and transcript abundances. Surface water concentrations of THg in Berry's Creek estuary (210-6800 pM) are among the highest in North America. A downstream gradient of Hg contamination is a permanent feature of Berry's Creek estuary, and the upper estuary appears to be a perennial source of Hg to the lower estuary and the Hackensack River. MeHg concentrations in Berry's Creek surface waters ranged from 2 to 14 pM, with the highest concentrations occurring at a midestuary site 2 km downstream of the tide gate. The suspended particle phase dominated Hg and MeHg speciation throughout this system, accounting for > 90% of THg in Berry's Creek estuary and 35-94% of THg in the Hackensack River. Concentrations of DGM in Berry's Creek estuary (0.1-1.0 pM) are similar to levels of DGM in other much less contaminated estuaries (0.04-0.75 pM). In addition, expression levels of the bacterial mercuric reductase gene, merA, a gene of the inorganic Hg(II)-regulated, mercury resistance (mer) operon, were low throughout Berry's Creek estuary. Thus, despite very high concentrations of mercury in Berry's Creek estuary, relatively low concentrations of DGM and merA gene expression levels indicate limited bioavailability of inorganic Hg in the estuary's surface waters. A system-wide limitation on the bioavailability of inorganic Hg, together with bacterial demethylation activity, may account for observed MeHg concentrations that, although elevated, are lower than expected given the concentrations of THg in this estuary.

High diversity of bacterial mercuric reductase genes from surface and sub-surface floodplain soil (Oak Ridge, USA)

DNA was extracted from different depth soils (0-5, 45-55 and 90-100 cm below surface) sampled at Lower East Fork Poplar Creek floodplain (LEFPCF), Oak Ridge (TN, USA). The presence of merA genes, encoding the mercuric reductase, the key enzyme in detoxification of mercury in bacteria, was examined by PCR targeting Actinobacteria, Firmicutes or beta/gamma-Proteobacteria. beta/gamma-Proteobacteria merA genes were successfully amplified from all soils, whereas Actinobacteria were amplified only from surface soil. merA clone libraries were constructed and sequenced. beta/gamma-Proteobacteria sequences revealed high diversity in all soils, but limited vertical similarity. Less than 20% of the operational taxonomic units (OTU) (DNA sequences > or = 95% identical) were shared between the different soils. Only one of the 62 OTU was > or = 95% identical to a GenBank sequence, highlighting that cultivated bacteria are not representative of what is found in nature. Fewer merA sequences were obtained from the Actinobacteria, but these were also diverse, and all were different from GenBank sequences. A single clone was most closely related to merA of alpha-Proteobacteria. An alignment of putative merA genes of genome sequenced mainly marine alpha-Proteobacteria was used for design of merA primers. PCR amplification of soil alpha-Proteobacteria isolates and sequencing revealed that they were very different from the genome-sequenced bacteria (only 62%-66% identical at the amino-acid level), although internally similar. In light of the high functional diversity of mercury resistance genes and the limited vertical distribution of shared OTU, we discuss the role of horizontal gene transfer as a mechanism of bacterial adaptation to mercury.

Analysis of mercuric reductase (merA) gene diversity in an anaerobic mercury‐contaminated sediment enrichment

The reduction of ionic mercury to elemental mercury by the mercuric reductase (MerA) enzyme plays an important role in the biogeochemical cycling of mercury in contaminated environments by partitioning mercury to the atmosphere. This activity, common in aerobic environments, has rarely been examined in anoxic sediments where production of highly toxic methylmercury occurs. Novel degenerate PCR primers were developed which span the known diversity of merA genes in Gram-negative bacteria and amplify a 285 bp fragment at the 3' end of merA. These primers were used to create a clone library and to analyse merA diversity in an anaerobic sediment enrichment collected from a mercury-contaminated site in the Meadowlands, New Jersey. A total of 174 sequences were analysed, representing 71 merA phylotypes and four novel MerA clades. This first examination of merA diversity in anoxic environments suggests an untapped resource for novel merA sequences.