Dataset Information

Cloning of merA Gene from Methylotenera Mobilis for Mercury Biotransformation.

ABSTRACT: Mercury (Hg) is one of the most toxic heavy metal and is extremely harmful for the environment. The permissible limit of mercury in industrial effluents is 0.001 ppm, whereas there are various sites having very high levels of mercury contamination. In the present study, 10 different mercury (Hg) resistant bacterial strains were isolated from Ulhas Estuary, Mumbai (Hg concentration of 107 ppm). All the strains were subsequently grown on higher concentration of mercuric chloride (HgCl₂), one of the isolate (USP5) showed significant growth at high concentration of Hg (40 ppm) and 16S rRNA gene sequencing revealed the identity of the bacterium as Methylotenera mobilis, (Accession no. KT714144). The mer operon was isolated and cloned in E.coli and checked for its ability to tolerate higher concentration of Hg. It has shown growth up to 70 ppm of Hg, also presence of merA gene indicated its ability to detoxify Hg into less toxic volatile form. The atomic absorption spectrophotometry confirmed the ability of clone to efficiently detoxify 60-90 % of the Hg (10-70 ppm) within 48-72 h. This clone can be used for effective volatilization of Hg from contaminated areas.

SUBMITTER: Porwal S

PROVIDER: S-EPMC5061702 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature

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Cloning of merA Gene from Methylotenera Mobilis for Mercury Biotransformation.

Porwal Shalini S Singh Rajni R

Indian journal of microbiology 20160728 4

Mercury (Hg) is one of the most toxic heavy metal and is extremely harmful for the environment. The permissible limit of mercury in industrial effluents is 0.001 ppm, whereas there are various sites having very high levels of mercury contamination. In the present study, 10 different mercury (Hg) resistant bacterial strains were isolated from Ulhas Estuary, Mumbai (Hg concentration of 107 ppm). All the strains were subsequently grown on higher concentration of mercuric chloride (HgCl2) ...[more]

PMID: 27784949

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Project description:Nine polymorphic mer loci carried by 185 gram-negative fecal bacterial strains from humans and nonhuman primates are described. The loci were characterized with specific intragenic and intergenic PCR primers to amplify distinct regions covering approximately 80% of the typical gram-negative mer locus. These loci were grouped phylogenetically with respect to each other and with respect to seven previously sequenced mer operons from gram-negative bacteria (the latter designated loci 1, 2, 3, 6, 7, 8, and delta 8 by us here for the purpose of this analysis). Six of the mer loci recovered from primates are similar either to these previously sequenced mer loci or to another locus recently observed in environmental isolates (locus 4), and three are novel (loci 5, 9, and 10). We have observed merC, or a merC-like gene, or merF on the 5' side of merA in all of the loci except that of Tn501 (here designated mer locus 6). The merB gene was observed occasionally, always on the 3' side of merA. Unlike the initial example of a merB-containing mer locus carried by plasmid pDU1358 (locus 8), all the natural primate loci carrying merB also had large deletions of the central region of the operon (and were therefore designated locus delta 8). Four of the loci we describe (loci 2, 5, 9, and 10) have no region of homology to merB from pDU1358 and yet strains carrying them were phenylmercury resistant. Two of these loci (loci 5 and 10) also lacked merD, the putative secondary regulator of operon expression. Phylogenetic comparison of character states derived from PCR product data grouped those loci which have merC into one clade; these are locus 1 (including Tn21), locus 3, and locus 4. The mer loci which lack merC grouped into a second clade: locus 6 (including Tn501) and locus 2. Outlying groups lacked merD or possessed merB. While these mer operons are characterized by considerable polymorphism, our ability to discern coherent clades suggests that recombination is not entirely random and indeed may be focused on the immediate 5' and 3' proximal regions of merA. Our observations confirm and extend the idea that the mer operon is a genetic mosaic and has a predominance of insertions and/or deletions of functional genes immediately before and after the merA gene. chi sites are found in several of the sequenced operons and may be involved in the abundant reassortments we observe for mer genes.

Project description:Mercury (Hg) is a highly toxic element due to its high affinity for protein sulfhydryl groups, which upon binding, can destabilize protein structure and decrease enzyme activity. Prokaryotes have evolved enzymatic mechanisms to detoxify inorganic Hg and organic Hg (e.g., MeHg) through the activities of mercuric reductase (MerA) and organomercury lyase (MerB), respectively. Here, the taxonomic distribution and evolution of MerAB was examined in 84,032 archaeal and bacterial genomes, metagenome assembled genomes, and single-cell genomes. Homologs of MerA and MerB were identified in 7.8 and 2.1% percent of genomes, respectively. MerA was identified in the genomes of 10 archaeal and 28 bacterial phyla previously unknown to code for this functionality. Likewise, MerB was identified in 2 archaeal and 11 bacterial phyla previously unknown to encode this functionality. Surprisingly, homologs of MerB were identified in a number of genomes (∼50% of all MerB-encoding genomes) that did not encode MerA, suggesting alternative mechanisms to detoxify Hg(II) once it is generated in the cytoplasm. Phylogenetic reconstruction of MerA place its origin in thermophilic Thermoprotei (Crenarchaeota), consistent with high levels of Hg(II) in geothermal environments, the natural habitat of this archaeal class. MerB appears to have been recruited to the mer operon relatively recently and likely among a mesophilic ancestor of Euryarchaeota and Thaumarchaeota. This is consistent with the functional dependence of MerB on MerA and the widespread distribution of mesophilic microorganisms that methylate Hg(II) at lower temperature. Collectively, these results expand the taxonomic and ecological distribution of mer-encoded functionalities, and suggest that selection for Hg(II) and MeHg detoxification is dependent not only on the availability and type of mercury compounds in the environment but also the physiological potential of the microbes who inhabit these environments. The expanded diversity and environmental distribution of MerAB identify new targets to prioritize for future research.

Dataset Information

Cloning of merA Gene from Methylotenera Mobilis for Mercury Biotransformation.

Publications

Cloning of <i>mer</i>A Gene from <i>Methylotenera Mobilis</i> for Mercury Biotransformation.

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