title: Characterization of mercury bioremediation by transgenic bacteria expressing metallothionein and polyphosphate kinase.
abstract: BACKGROUND
  The use of transgenic bacteria has been proposed as a suitable alternative for mercury remediation. Ideally, mercury would be sequestered by metal-scavenging agents inside transgenic bacteria for subsequent retrieval. So far, this approach has produced limited protection and accumulation. We report here the development of a transgenic system that effectively expresses metallothionein (mt-1) and polyphosphate kinase (ppk) genes in bacteria in order to provide high mercury resistance and accumulation.
  
  
  RESULTS
  In this study, bacterial transformation with transcriptional and translational enhanced vectors designed for the expression of metallothionein and polyphosphate kinase provided high transgene transcript levels independent of the gene being expressed. Expression of polyphosphate kinase and metallothionein in transgenic bacteria provided high resistance to mercury, up to 80 μM and 120 μM, respectively. Here we show for the first time that metallothionein can be efficiently expressed in bacteria without being fused to a carrier protein to enhance mercury bioremediation. Cold vapor atomic absorption spectrometry analyzes revealed that the mt-1 transgenic bacteria accumulated up to 100.2 ± 17.6 μM of mercury from media containing 120 μM Hg. The extent of mercury remediation was such that the contaminated media remediated by the mt-1 transgenic bacteria supported the growth of untransformed bacteria. Cell aggregation, precipitation and color changes were visually observed in mt-1 and ppk transgenic bacteria when these cells were grown in high mercury concentrations.
  
  
  CONCLUSIONS
  The transgenic bacterial system described in this study presents a viable technology for mercury bioremediation from liquid matrices because it provides high mercury resistance and accumulation while inhibiting elemental mercury volatilization. This is the first report that shows that metallothionein expression provides mercury resistance and accumulation in recombinant bacteria. The high accumulation of mercury in the transgenic cells could present the possibility of retrieving the accumulated mercury for further industrial applications.
authors.affiliation: Inter American University of Puerto Rico, Department of Natural Sciences and Mathematics, 500 Dr. John Will Harris, Bayamon, Puerto Rico. Oscar.Ruiz@wpafb.af.mil
authors.forename: Oscar N
authors.initials: ON
authors.lastname: Ruiz
authors.forename: Derry
authors.initials: D
authors.lastname: Alvarez
authors.forename: Gloriene
authors.initials: G
authors.lastname: Gonzalez-Ruiz
authors.forename: Cesar
authors.initials: C
authors.lastname: Torres
mesh_terms: D001673:Biodegradation, Environmental; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D008628:Mercury / Q000493:pharmacokinetics*; D008668:Metallothionein / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D019076:Transgenes
chemical_list: D029968:Escherichia coli Proteins; C525355:metallothionein-1, mouse; D008668:Metallothionein; D017853:Phosphotransferases (Alcohol Group Acceptor); C475691:polyphosphate kinase, E coli; D008628:Mercury