Heavy metal resistance; CH34; plasmids; Alcaligenes eutrophus
Abstract :
[en] Alcaligenes eutrophus CH34 is the main representative of a group of strongly related strains (mostly facultative chemolithotrophs) that are well adapted to environments containing high levels of heavy metals. It harbors the megaplasmids pMOL28 and pMOL30 which carry resistance determinants to Co2+, Ni2+, CrO42-, Hg2+, Tl+, Cd2+, Cu2+ and Zn2+. Among the best characterized determinants are the cnr operon (resistance to Co, Ni) an pMOL28 and the czc operon on pMOL30 (resistance to Co, Cd and Zn). Although the two systems reveal a significant degree of amino acid similarity in the structural genes, the regulation of the operons is different. The resistance mechanism in both cases is based on efflux. The efflux mechanism leads to a pH increase outside of the cytoplasmic membrane. Metals are sequestered from the external medium through the bioprecipitation of metal carbonates formed in the saturated zone around the cell. This latter phenomenon can be exploited in bioreactors designed to remove metals from effluents. The bacteria are immobilized on composite membranes in a continuous tubular membrane reactor (CTMR). The effluent continuously circulates through the intertubular space, while the external surface of the tubes is in contact with the growth medium. Metal crystals are eventually removed by the effluent stream and collected on a glass bead column. The system has been applied to effluents containing Cd, Zn, Co, Ni and Cu. By introducing catabolic plasmids involved in the aerobic degradation of PCBs and 2,4-D into metal-resistant A. eutrophus strains, the application range was widened to include effluents polluted with both organic and inorganic substances. Biosensors have been developed which are based on the fusion of genes induced by metals to a reporter system, the lux operon of Vibrio fischeri. Bacterial luciferases produce light through the oxidation of fatty aldehydes. The gene fusions are useful both for the study of regulatory genes and for the determination of heavy metal concentrations in the environment.
Disciplines :
Microbiology
Author, co-author :
Collard, Jean-Marc
Corbisier, Philippe
Diels, Ludo
Dong, Q.
Jeanthon, Christian
Mergeay, Max ; Université de Liège - ULiège > Département Argenco : Secteur GeMMe > Génie minéral et recyclage
Taghavi, S.
Van Der Lelie, Daniel
Wilmotte, Annick ; Université de Liège - ULiège > Département des sciences de la vie > Physiologie et génétique bactériennes
Wuertz, Stefan
Language :
English
Title :
Plasmids for heavy metal resistance in Alcaligenes eutrophus CH34: Mechanisms and applications
Novick, Roth, Plasmid-linked resistance to inorganic salts in Staphylococcus aureus. (1968) J Bacteriol, 95, pp. 1335-1342
Silver, Budd, Leahy, Shaw, Hammond, Novick, Willsky, Rosenberg, Inducible plasmid-determined resistance to arsenate, arsenite and antimony (III) in Escherichia coli and Staphylococcus aureus (1981) J. Bacteriol., 146, pp. 983-996
Summers, Organization, expression and evolution of genes for mercury resistance (1986) Annu. Rev. Microbiol., 40, pp. 607-634
Silver, Misra, Plasmid-mediated heavy metal resistance (1988) Annu. Rev. Microbiol., 42, pp. 717-743
Kaur, Rosen, Plasmid-encoded resistance to arsenic and antimony (1992) Plasmid, 27, pp. 29-40
Silver, Walderhaug, Gene regulation of plasmid- and chromosome-determined inorganic ion transport in bacteria (1992) Microbiol. Rev., 56, pp. 195-228
Brown, Rouch, Lee, Copper resistance systems in bacteria (1992) Plasmid, 27, pp. 41-51
Cooksey, Copper uptake and resistance in bacteria (1993) Mol. Microbiol., 7, pp. 1-5
Mergeay, Nies, Schlegel, Gerits, Charles, Van Gijsegem, Alcaligenes eutrophus CH34 is a facultative chemolithotroph with plasmid bound resistance to heavy metals (1985) J. Bacteriol., 162, pp. 328-334
Diels, Mergeay, DNA probe mediated detection of resistant bacteria from soils highly polluted by heavy metals (1990) Appl. Environ. Microbiol., 56, pp. 1485-1491
Schmidt, Stoppel, Schlegel, High-level nickel resistance in Alcaligenes xylosoxydans 31A and Alcaligenes eutrophus KTO2 (1991) Appl. Environ. Microbiol., 57, pp. 3301-3309
Kortlüke, Hogrefe, Eberz, Pühler, Friedrich, Genes of lithoautotrophic metabolism are clustered on the megaplasmid pHG1 in Alcaligenes eutrophus (1987) MGG Molecular & General Genetics, 210, pp. 122-128
Sadouk, Mergeay, Chromosome mapping in Alcaligenes eutrophus CH34 (1993) Mol. Gen. Genet., 240, pp. 181-187
Nies, Nies, Chu, Silver, Expression and nucleotide sequence of u plasmid-determined divalent cation efflux system from Alcaligenes eutrophus (1989) Proc. Natl. Acad. Sci. USA, 86, pp. 7351-7355
Nies, Silver, Plasmid-dctermined inducible efflux is responsible for resistance to cadmium, zinc and cobalt in Alcaligenes eutrophus (1989) J. Bacteriol., 171, pp. 896-900
Nies, czcR and czcD, gene products affecting regulation of resistance to cobalt, zinc and cadmium (czc system) in Alcaligenes eutrophus (1992) J. Bacteriol., 174, pp. 8102-8110
Liesegang, Lemke, Siddiqui, Schlegel, Characterization of the inducible nickel and cobalt resistance determinant cnr from pMOL28 of Alcaligenes eutrophus CH34 (1993) J. Bacteriol., 175, pp. 767-778
Lonetto, Brown, Rudd, Buttner, Analysis of the Streptomyces coelicolor sigE gene reveals the existence of a subfamily of eubacterial RNA polymerase sigma factors involved in the regulation of extracytoplasmic functions. (1994) Proc Natl Acad Sci U S A, , in press
Collard, Provoost, Taghavi, Mergeay, A new type of Alcaligenes eutrophus CH34 zinc resistance generated by mutations affecting regulation of the cnr cobalt-nickel resistance system (1993) J. Bacteriol., 175, pp. 779-784
Nies, Nies, Silver, Nucleotide sequence and expression of a plasmid-encoded chromate resistance determination from Alcaligenes eutrophus (1990) J. Biol. Chem., 265, pp. 5648-5653
Diels, Sadouk, Mergeay, Large plasmids governing multiple resistances to heavy metals: A genetic approach∗ (1989) Toxicological & Environmental Chemistry, 23, pp. 79-89
Mergeay, Towards an understanding of the genetics of bacterial metal resistance (1991) Trends Biotechnol., 9, pp. 17-24
Diels, Van Roy, Mergeay, Doyen, Taghavi, Leysen, Immobilization of bacteria in composite membranes and development of tubular membrane reactors for heavy metal recuperation. New Perspectives (1993) Effective Membrane Processes, , T. Peters, Kluwer Academic Publishers, Dordrecht
Diels, Van Roy, Taghavi, Doyen, Leysen, Mergeay, (1993) The use of Alcaligenes eutrophus immobilized in a tubular membrane reactor for heavy metal recuperation, , Biohydrometallurgy Meeting, Jackson Hole, August 22–25, 1993
Springael, Kreps, Mergeay, Identification of a catabolic transposon, Tn437l, carrying biphenyl and 4-chlorobiphenyl degradation genes in Alcaligenes eutrophus A5 (1993) J. Bacteriol., 175, pp. 1674-1681
Springael, Diels, Hooyberghs, Kreps, Mergeay, Construction and characterization of heavy metal-resistant haloaromatic-degrading Alcaligenes eutrophus strains (1993) Appl. Environ. Microbiol., 59, pp. 334-339
Van der Lelie, Corbisier, Baeyens, Wuertz, Diels, Mergeay, The use of biosensors for environmental monitoring (1994) Res. Microbiol., 145, pp. 67-74
Corbisier, Ji, Nuyts, Mergeay, Silver, luxAB gene fusions with the arsenic and cadmium resistance operons of Staphylococcus aureus plasmid pI258 (1993) FEMS Microbiol. Lett., 110, pp. 231-238
Top, Mergeay, Springael, Verstraete, Gene escape model: transfer ot heavy metal resistance genes from Escherichia coli to Alcaligenes eutrophus on agar plates and in soil samples (1990) Appl. Environ. Microbiol., 56, pp. 2471-2479