Applied Microbiology and Biotechnology

, Volume 87, Issue 1, pp 53–60

Engineering of microorganisms towards recovery of rare metal ions



The bioadsorption of metal ions using microorganisms is an attractive technology for the recovery of rare metal ions as well as removal of toxic heavy metal ions from aqueous solution. In initial attempts, microorganisms with the ability to accumulate metal ions were isolated from nature and intracellular accumulation was enhanced by the overproduction of metal-binding proteins in the cytoplasm. As an alternative, the cell surface design of microorganisms by cell surface engineering is an emerging strategy for bioadsorption and recovery of metal ions. Cell surface engineering was firstly applied to the construction of a bioadsorbent to adsorb heavy metal ions for bioremediation. Cell surface adsorption of metal ions is rapid and reversible. Therefore, adsorbed metal ions can be easily recovered without cell breakage, and the bioadsorbent can be reused or regenerated. These advantages are suitable for the recovery of rare metal ions. Actually, the cell surface display of a molybdate-binding protein on yeast led to the enhanced adsorption of molybdate, one of the rare metal ions. An additional advantage is that the cell surface display system allows high-throughput screening of protein/peptide libraries owing to the direct evaluation of the displayed protein/peptide without purification and concentration. Therefore, the creation of novel metal-binding protein/peptide and engineering of microorganisms towards the recovery of rare metal ions could be simultaneously achieved.


Rare metals Metal recovery Bioadsorption Cell surface engineering Arming yeast 


  1. Akthar N, Sastry S, Mohan M (1995) Biosorption of silver ions by processed Aspergillus niger biomass. Biotechnol Lett 17:551–556CrossRefGoogle Scholar
  2. Anonymous (1997) Arming yeast with cell-surface catalysts. Chem Eng News 75:32Google Scholar
  3. Bae W, Chen W, Mulchandani A, Mehra RK (2000) Enhanced bioaccumulation of heavy metals by bacterial cells displaying synthetic phytochelatins. Biotechnol Bioeng 70:518–524CrossRefGoogle Scholar
  4. Bae W, Wu CH, Kostal J, Mulchandani A, Chen W (2003) Enhanced mercury biosorption by bacterial cells with surface-displayed MerR. Appl Environ Microbiol 69:3176–3180CrossRefGoogle Scholar
  5. Berka T, Shatzman A, Zimmerman J, Strickler J, Rosenberg M (1988) Efficient expression of the yeast metallothionein gene in Escherichia coli. J Bacteriol 170:21–26Google Scholar
  6. Brown S (1997) Metal-recognition by repeating polypeptides. Nat Biotechnol 15:269–272CrossRefGoogle Scholar
  7. Clemens S (2001) Molecular mechanisms of plant metal tolerance and homeostasis. Planta 212:475–486CrossRefGoogle Scholar
  8. Dong J, Liu C, Zhang J, Xin ZT, Yang G, Gao B, Mao CQ, Liu NL, Wang F, Shao NS, Fan M, Xue YN (2006) Selection of novel nickel-binding peptides from flagella displayed secondary peptide library. Chem Biol Drug Des 68:107–112CrossRefGoogle Scholar
  9. Eccles H (1999) Treatment of metal-contaminated wastes: why select a biological process? Trends Biotechnol 17:462–465CrossRefGoogle Scholar
  10. Eide DJ (1998) The molecular biology of metal ion transport in Saccharomyces cerevisiae. Annu Rev Nutr 18:441–469CrossRefGoogle Scholar
  11. Gadd GM, White C (1993) Microbial treatment of metal pollution—a working biotechnology? Trends Biotechnol 11:353–359CrossRefGoogle Scholar
  12. Georgiou G, Poetschke HL, Stathopoulos C, Francisco JA (1993) Practical applications of engineering Gram-negative bacterial cell surfaces. Trends Biotechnol 11:6–10CrossRefGoogle Scholar
  13. Georgiou G, Stathopoulos C, Daugherty PS, Nayak AR, Iverson BL, Curtiss R 3rd (1997) Display of heterologous proteins on the surface of microorganisms: from the screening of combinatorial libraries to live recombinant vaccines. Nat Biotechnol 15:29–34CrossRefGoogle Scholar
  14. Hall JL (2002) Cellular mechanisms for heavy metal detoxification and tolerance. J Exp Bot 53:1–11CrossRefGoogle Scholar
  15. Holm RH, Kennepohl P, Solomon EI (1996) Structural and functional aspects of metal sites in biology. Chem Rev 96:2239–2314CrossRefGoogle Scholar
  16. Hou YM, Kim R, Kim SH (1988) Expression of the mouse metallothionein-I gene in Escherichia coli: increased tolerance to heavy metals. Biochim Biophys Acta 951:230–234Google Scholar
  17. Kapoor A, Viraraghavan T (1995) Fungal biosorption—an alternative treatment option for heavy metal bearing wastewaters: A review. Bioresour Technol 53:195–206CrossRefGoogle Scholar
  18. Khoo K, Ting Y (2001) Biosorption of gold by immobilized fungal biomass. Biochem Eng J 8:51–59CrossRefGoogle Scholar
  19. Kisker C, Schindelin H, Rees DC (1997) Molybdenum-cofactor-containing enzymes: structure and mechanism. Annu Rev Biochem 66:233–267CrossRefGoogle Scholar
  20. Kjaergaard K, Schembri MA, Klemm P (2001) Novel Zn2+-chelating peptides selected from a fimbria-displayed random peptide library. Appl Environ Microbiol 67:5467–5473CrossRefGoogle Scholar
  21. Kondo A, Ueda M (2004) Yeast cell-surface display—applications of molecular display. Appl Microbiol Biotechnol 64:28–40CrossRefGoogle Scholar
  22. Kotrba P, Doleckova L, de Lorenzo V, Ruml T (1999) Enhanced bioaccumulation of heavy metal ions by bacterial cells due to surface display of short metal binding peptides. Appl Environ Microbiol 65:1092–1098Google Scholar
  23. Kurniawan TA, Chan GYS, Lo WH, Babel S (2006) Physico-chemical treatment techniques for wastewater laden with heavy metals. Chem Eng J 118:83–98Google Scholar
  24. Kuroda K, Ueda M (2003) Bioadsorption of cadmium ion by cell surface-engineered yeasts displaying metallothionein and hexa-His. Appl Microbiol Biotechnol 63:182–186CrossRefGoogle Scholar
  25. Kuroda K, Ueda M (2006) Effective display of metallothionein tandem repeats on the bioadsorption of cadmium ion. Appl Microbiol Biotechnol 70:458–463CrossRefGoogle Scholar
  26. Kuroda K, Shibasaki S, Ueda M, Tanaka A (2001) Cell surface-engineered yeast displaying a histidine oligopeptide (hexa-His) has enhanced adsorption of and tolerance to heavy metal ions. Appl Microbiol Biotechnol 57:697–701CrossRefGoogle Scholar
  27. Kuroda K, Ueda M, Shibasaki S, Tanaka A (2002) Cell surface-engineered yeast with ability to bind, and self-aggregate in response to, copper ion. Appl Microbiol Biotechnol 59:259–264CrossRefGoogle Scholar
  28. Ledin M (2000) Accumulation of metals by microorganisms—processes and importance for soil systems. Earth-Sci Rev 51:1–31CrossRefGoogle Scholar
  29. Lee SY, Choi JH, Xu Z (2003) Microbial cell-surface display. Trends Biotechnol 21:45–52CrossRefGoogle Scholar
  30. Lovley DR, Coates JD (1997) Bioremediation of metal contamination. Curr Opin Biotechnol 8:285–289CrossRefGoogle Scholar
  31. Matsui K, Kuroda K, Ueda M (2009) Creation of a novel peptide endowing yeasts with acid tolerance using yeast cell-surface engineering. Appl Microbiol Biotechnol 82:105–113CrossRefGoogle Scholar
  32. Mejare M, Ljung S, Bulow L (1998) Selection of cadmium specific hexapeptides and their expression as OmpA fusion proteins in Escherichia coli. Protein Eng 11:489–494CrossRefGoogle Scholar
  33. Nishitani T, Shimada M, Kuroda K, Ueda M (2010) Molecular design of yeast cell surface for adsorption and recovery of molybdenum, one of rare metals. Appl Microbiol Biotechnol 86:641–648CrossRefGoogle Scholar
  34. Nriagu JO, Pacyna JM (1988) Quantitative assessment of worldwide contamination of air, water and soils by trace metals. Nature 333:134–139CrossRefGoogle Scholar
  35. Pazirandeh M, Chrisey LA, Mauro JM, Campbell JR, Gaber BP (1995) Expression of the Neurospora crassa metallothionein gene in Escherichia coli and its effect on heavy-metal uptake. Appl Microbiol Biotechnol 43:1112–1117CrossRefGoogle Scholar
  36. Perego P, Howell SB (1997) Molecular mechanisms controlling sensitivity to toxic metal ions in yeast. Toxicol Appl Pharmacol 147:312–318CrossRefGoogle Scholar
  37. Pethkar AV, Kulkarni SK, Paknikar KM (2001) Comparative studies on metal biosorption by two strains of Cladosporium cladosporioides. Bioresour Technol 80:211–215CrossRefGoogle Scholar
  38. Reddy BR, Priya DN (2004) Solvent extraction of Ni(II) from sulfate solutions with LIX 84I: flow-sheet for the separation of Cu(II), Ni(II) and Zn(II). Anal Sci 20:1737–1740CrossRefGoogle Scholar
  39. Saleem M, Brim H, Hussain S, Arshad M, Leigh MB, Zia-ul H (2008) Perspectives on microbial cell surface display in bioremediation. Biotechnol Adv 26:151–161CrossRefGoogle Scholar
  40. Samuelson P, Wernerus H, Svedberg M, Stahl S (2000) Staphylococcal surface display of metal-binding polyhistidyl peptides. Appl Environ Microbiol 66:1243–1248CrossRefGoogle Scholar
  41. Samuelson P, Gunneriusson E, Nygren PA, Stahl S (2002) Display of proteins on bacteria. J Biotechnol 96:129–154CrossRefGoogle Scholar
  42. Savvaidis I (1998) Recovery of gold from thiourea solutions using microorganisms. Biometals 11:145–151CrossRefGoogle Scholar
  43. Sayers Z, Brouillon P, Vorgias CE, Nolting HF, Hermes C, Koch MH (1993) Cloning and expression of Saccharomyces cerevisiae copper-metallothionein gene in Escherichia coli and characterization of the recombinant protein. Eur J Biochem 212:521–528CrossRefGoogle Scholar
  44. Self WT, Grunden AM, Hasona A, Shanmugam KT (2001) Molybdate transport. Res Microbiol 152:311–321CrossRefGoogle Scholar
  45. Sousa C, Kotrba P, Ruml T, Cebolla A, De Lorenzo V (1998) Metalloadsorption by Escherichia coli cells displaying yeast and mammalian metallothioneins anchored to the outer membrane protein LamB. J Bacteriol 180:2280–2284Google Scholar
  46. Tsuruta T (2004) Biosorption and recycling of gold using various microorganisms. J Gen Appl Microbiol 50:221–228CrossRefGoogle Scholar
  47. Ueda M (2004) Future direction of molecular display by yeast-cell surface engineering. J Mol Catal B 28:139–143CrossRefGoogle Scholar
  48. Ueda M, Tanaka A (2000a) Cell surface engineering of yeast: construction of arming yeast with biocatalyst. J Biosci Bioeng 90:125–136Google Scholar
  49. Ueda M, Tanaka A (2000b) Genetic immobilization of proteins on the yeast cell surface. Biotechnol Adv 18:121–140CrossRefGoogle Scholar
  50. Vasudevan P, Padmavathy V, Dhingra SC (2002) Biosorption of monovalent and divalent ions on baker's yeast. Bioresour Technol 82:285–289CrossRefGoogle Scholar
  51. Vijayaraghavan K, Jegan J, Palanivelu K, Velan M (2004) Removal of nickel(II) ions from aqueous solution using crab shell particles in a packed bed up-flow column. J Hazard Mater 113:223–230CrossRefGoogle Scholar
  52. Volesky B, May-Phillips HA (1995) Biosorption of heavy metals by Saccharomyces cerevisiae. Appl Microbiol Biotechnol 42:797–806CrossRefGoogle Scholar
  53. Volesky B, Weber J, Park JM (2003) Continuous-flow metal biosorption in a regenerable Sargassum column. Water Res 37:297–306CrossRefGoogle Scholar
  54. Wagner UG, Stupperich E, Kratky C (2000) Structure of the molybdate/tungstate binding protein mop from Sporomusa ovata. Structure 8:1127–1136CrossRefGoogle Scholar
  55. Wang HL, Chen C (2006) Biosorption of heavy metals by Saccharomyces cerevisiae: a review. Biotechnol Adv 24:427–451CrossRefGoogle Scholar
  56. Wittrup KD (2001) Protein engineering by cell-surface display. Curr Opin Biotechnol 12:395–399CrossRefGoogle Scholar
  57. Yong P, Rowson NA, Farr JPG, Harris IR, Macaskie LE (2002) Bioaccumulation of palladium by Desulfovibrio desulfuricans. J Chem Technol Biotechnol 77:593–601CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Division of Applied Life Sciences, Graduate School of AgricultureKyoto UniversityKyotoJapan

Personalised recommendations