1887

Microbiology Society logo

Volume 143, Issue 7

Localization of enzymically enhanced heavy metal accumulation by sp. and metal accumulation by liposomes containing entrapped enzyme Free

Abstract

Summary: A heavy-metal-accumulating sp. has been used for the treatment of metal-laden industrial wastes. Metal uptake is mediated via a cell-bound phosphatase that liberates inorganic phosphate which precipitates with heavy metals as cell-bound metal phosphate. A phosphatase-deficient mutant accumulated little UO , while a phosphatase-overproducing mutant accumulated correspondingly more metal, with a uranium loading equivalent to the bacterial dry weight achieved after 6 h exposure of resting cells to uranyl ion in the presence of phosphatase substrate (glycerol 2-phosphate). The phosphatase, visualized by immunogold labelling in the parent and overproducing strains, but not seen in the deficient mutant, was held within the periplasmic space with, in some cells, a higher concentration at the polar regions. Enzyme was also associated with the outer membrane and found extracellularly. Accumulated uranyl phosphate was visible as cell-surface- and polar-localized deposits, identified by energy-dispersive X-ray analysis (EDAX), proton-induced X-ray emission analysis (PIXE) and X-ray diffraction analysis (XRD) as polycrystalline HUOPO.4HO. Nuclaation sites for initiation of biocrystallization were identified at the cytoplasmic and outer membranes, prompting consideration of an biocatalytic system for metal waste remediation. Phosphatidylcholine-based liposomes with entrapped phosphatase released phosphate comparably to whole cells, as shown by 31P NMR spectroscopy in the presence of ‘IMMR-silent’ Cd. Application of liposome-immobilized enzyme to the decontamination of uranyl solutions was, however, limited by rapid fouling of the biocatalyst by deposited uranyl phosphate. It is suggested that the architecture of the bacterial cell surface provides a means of access of uranyl ion to the inner and outer membranes and enzymically liberated phosphate in a way that minimizes fouling in whole cells.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): Citrobacter sp. , liposomes , phosphatase and uranium uptake
Funding
This study was supported by the:
  • EU (Award BR2/0199/C)
  • Kyunggi Do and KOSEF
  • EU (Award BR2/0199/C)
© Society for General Microbiology 1997
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-7-2497
1997-07-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/7/mic-143-7-2497.html?itemId=/content/journal/micro/10.1099/00221287-143-7-2497&mimeType=html&fmt=ahah

References

  1. Barnes L. J., Janssen F. J., Sherran J., Versteegh J. H., Koch R. O., Scheeren P. J. H. 1991; A new process for the microbial removal of sulfate and heavy metals from contaminated wastes extracted by a geohydrological control system. Chem Eng Res Des 69A:184–186
     [Google Scholar]
  2. Barnes L. J., Janssen F. J., Scheeren P., Versteegh J. H., Koch R. O. 1992; Simultaneous microbial removal of sulphate and heavy metals from waste water. Trans Inst Mining Metallurg C 101:183–189
     [Google Scholar]
  3. Bonthrone K. M., Basnakova G., Lin F., Macaskie L. E. 1996; Bioaccumulation of nickel by intercalation into polycrystalline hydrogen uranyl phosphate deposited via an enzymatic mechanism. Nat Biotechnol 14:635–638
     [Google Scholar]
  4. Butler A. J., Hallett D. S., Macaskie L. E. 1991; Phosphatase production by a Citrobacter sp. growing in batch culture and use of batch cultures to investigate some limitations in the use of polyacrylamide gel-immobilized cells for product release. Enzyme Microb Technol 13:716–721
     [Google Scholar]
  5. Diels L, Dong Q., Van der Lelie D., Baeyens W., Mergeay M. 1995; The czc operon of Alcaligenes eutrophus CH34: from resistance mechanism to the removal of heavy metals. J Ind Microbiol 14:142–153
     [Google Scholar]
  6. Hughes M. N., Poole R. K. 1991; Metal speciation and microbial growth; the hard (and soft) facts. J Gen Microbiol 137:725–734
     [Google Scholar]
  7. Jeong B. C. 1992 Studies on the atypical phosphatase of a metal-accumulating Citrobacter sp DPhil thesis University of Oxford; UK:
     [Google Scholar]
  8. Jeong B. C., Macaskie L. E. 1995; PhoN-type acid phosphatases of a heavy metal-accumulating Citrobacter sp.: resistance to heavy metals and affinity towards phosphomonoester substrates. FEMS Microbiol Lett 130:211–214
     [Google Scholar]
  9. Jeong B. C., Kim H. W., Owen S. J., Dick R. E., Macaskie L. E. 1994; Phosphoesterase activity and phosphate release from tributyl phosphate by a Citrobacter sp. Appl Biochem Biotechnol 47:21–32
     [Google Scholar]
  10. Kier L. D., Weppelman R., Ames B. N. 1977; Resolution and purification of three periplasmic phosphatases of Salmonella typhimurium . J Bacteriol 130:399–410
     [Google Scholar]
  11. Lloyd J. R., Macaskie L. E. 1996; Bacterial reduction and removal of technetium from solution. Proceedings of the 3rd International Conference on Minerals Bioprocessing and Biorecovery/Bioremediation in Mining Big Sky, Montana, USA, 25–30 August 1996
     [Google Scholar]
  12. Lloyd J. R., Cole J. A., Macaskie L. E. 1997; Reduction and removal of heptavalent technetium from solution by Escherichia coli . J Bacteriol 179:2014–2021
     [Google Scholar]
  13. Macaskie L. E. 1990; An immobilized cell bioprocess for the removal of heavy metals from aqueous flows. J Chem Technol Biotechnol 49:357–379
     [Google Scholar]
  14. Macaskie L. E. 1995; Copper tolerance, phosphatase activity and copper uptake by a heavy metal-accumulating Citrobacter sp. Microbios 84:137–153
     [Google Scholar]
  15. Macaskie L. E., Blackmore J. D., Empson R. M. 1988; Phosphatase overproduction and enhanced uranium accumulation by a stable mutant of a Citrobacter sp. isolated by a novel method. FEMS Microbiol Lett 55:157–162
     [Google Scholar]
  16. Macaskie L. E., Empson R. M., Cheetham A. K., Grey C. P., Skarnulis A. J. 1992; Uranium bioaccumulation by a Citrobacter sp. as a result of enzymically-mediated growth of polycrystalline HUO2PO4 . Science 257:782–784
     [Google Scholar]
  17. Macaskie L. E., Jeong B. C., Tolley M. R. 1994a; Enzymically-accelerated biomineralization of heavy metals: application to the removal of americium and plutonium from aqueous flows. FEMS Microbiol Rev 14:351–368
     [Google Scholar]
  18. Macaskie L. E., Bonthrone K. M., Rouch D. A. 1994b; Phosphatase-mediated heavy metal accumulation by a Citrobacter sp. and related enterobacteria. FEMS Microbiol Lett 121:141–146
     [Google Scholar]
  19. Macaskie L. E., Empson R. M., Lin F., Tolley M. R. 1995a; Enzymatically-mediated uranium accumulation and uranium recovery using a Citrobacter sp. immobilized as a biofilm within a plug-flow reactor. J Chem Technol Biotechnol 63:1–16
     [Google Scholar]
  20. Macaskie L. E., Hewitt C. J., Shearer J. A., Kent C. A. 1995b; Biomass production for the removal of heavy metals from aqueous solution at low pH using growth-decoupled cells of a Citrobacter sp. Int Biodeterior Biodegrad 35:73–92
     [Google Scholar]
  21. Macaskie L. E., Lloyd J. R., Thomas R. A. P., Tolley M. R. 1996; The use of microorganisms for the remediation of solutions contaminated with actinide elements, other radionuclides, and organic contaminants generated by nuclear fuel cycle activities. Nucl Energy 35:257–271
     [Google Scholar]
  22. Macaskie L. E., Yong P., Doyle T. C., Roig M. G., Diaz M., Manzano T. 1997; Bioremediation of uranium-bearing wastewater: biochemical and chemical factors affecting bioprocess application. Biotechnol Bioeng 53:100–109
     [Google Scholar]
  23. Mann S., Hannington J. P., Williams R. J. P. 1986; Phospholipid vesicles as a model system for biomineralization. Nature 324:565–567
     [Google Scholar]
  24. Marques A. M., Roca X., Simon-Pujol M. D., Fuste M. C., Francisco C. 1991; Uranium accumulation by Pseudomonas sp. EPS-5028. Appl Microbiol Biotechnol 35:406–410
     [Google Scholar]
  25. Mergeay M., Nies D., Schlegel H. G., Gerits J., Van Gijsegen F. 1985; Alcaligenes eutrophus CH34, a facultative chemolithotroph displaying plasmid bound resistance to heavy metals. J Bacteriol 162:328–334
     [Google Scholar]
  26. Nesmeyanova M. A., Tsfasman I. M., Karamyshev A. L., Suzina N. E. 1991; Secretion of the overproduced periplasmic PhoA protein into the medium and accumulation of its precursor in phoA-transformed Escherichia coli strains: involvement of outer membrane vesicles. World J Microbiol Biotechnol 7:394–406
     [Google Scholar]
  27. Neu H. C., Heppel L. A. 1965; The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem 240:3685–3692
     [Google Scholar]
  28. Nossal N. G., Heppel L. A. 1966; The release of enzymes by osmotic shock from Escherichia coli in exponential phase. J Biol Chem 241:3055–3062
     [Google Scholar]
  29. Pierpoint W. S. 1957; The phosphatase and metaphosphatase activities of pea extracts. Biochem J 65:67–76
     [Google Scholar]
  30. Postgate J. R. 1979 The Sulphate Reducing Bacteria Cambridge: Cambridge University Press;
     [Google Scholar]
  31. Premuzic E. T., Francis A. J., Lin M., Schubert J. 1985; Induced formation of chelating agents by Pseudomonas aeruginosa grown in the presence of thorium and uranium. Arch Environ Contam Toxicol 14:759–768
     [Google Scholar]
  32. Roig M. G., Manzano T., Diaz M., Pascual M. J. 1995; Enzymically-yenhanced extraction of uranium from biologically-leached solutions. Int Biodeterior Biodegrad 35:93–128
     [Google Scholar]
  33. Strachan L. F., Jeong B. C., Macaskie L. E. 1991; Radiotolerance of phosphatases of a Citrobacter sp.: potential for the use of this organism in the treatment of wastes containing radiotoxic actinides. Proceedings of the 201st Meeting of the American Chemical Society Symposium: Biotechnology for Wastewater Treatment 31:128–131
     [Google Scholar]
  34. Strandberg G. W., Shumate S. E., II, Parrott J. R. 1981; Microbial cells as biosorbents for heavy metals: accumulation of uranium by Saccbaromyces cerevisiae and Pseudomonas aeruginosa. Appl Environ Microbiol 41:237–245
     [Google Scholar]
  35. Tolley M. R., Strachan L. F., Macaskie L. E. 1995; Lanthanum accumulation from acidic solutions using a Citrobacter sp. immobilized in a flow-through bioreactor. J Ind Microbiol 14:271–280
     [Google Scholar]
  36. Volesky B. 1990 Biosorption of Heavy Metals Boca Raton, FL: CRC Press;
     [Google Scholar]
  37. Weigel F. 1986; Uranium. . In The Chemistry of the Actinide Elements, 2nd edn, pp. 169–442 . Edited by Katz J. J., Seaborg G. T., Morss L. R. London: Chapman & Hall;
     [Google Scholar]
  38. Yong P., Macaskie L. E. 1995; Enhancement of uranium bioaccumulation by a Citrobacter sp. via enzymically-mediated growth of polycrystalline NH4UO2PO4 . J Chem Technol Bio-technol 63:101–108
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-7-2497
Loading
Localization of enzymically enhanced heavy metal accumulation by Citrobacter sp. and metal accumulation in vitro by liposomes containing entrapped enzyme
Microbiology 143, 2497 (1997); https://doi.org/10.1099/00221287-143-7-2497
/content/journal/micro/10.1099/00221287-143-7-2497
/content/journal/micro/10.1099/00221287-143-7-2497
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error