, 21:591 | Cite as

Chromate removal by yeasts isolated from sediments of a tanning factory and a mine site in Argentina

  • Liliana B. Villegas
  • Pablo M. Fernández
  • María J. Amoroso
  • Lucía I. C. de Figueroa


Twenty-one yeast-like microorganisms were isolated from tannery effluents and from a nickel–copper mine in Argentina. They were tested for their Cu(II), Ni(II), Cd(II) and Cr(VI) tolerance in qualitative assays on solid medium. Three isolates were selected for their multiple tolerance to the different heavy metals and highest tolerance to Cr(VI). According to morphological and physiological analysis and 26S rDNA D1/D2 domain sequences the isolates were characterized as: Lecythophora sp. NGV-1, Candida sp. NGV-9 and Aureobasidium pullulans VR-8. Resistance of the three strains to high Cr(VI) concentrations and their ability to remove Cr(VI) were assessed using YNB-glucose medium supplemented with 0.5 and 1 mM Cr(VI). Chromate removal activity was estimated by measuring remaining Cr(VI) concentration in the supernatant using the colorimetric 1,5-diphenylcarbazide method and total chromium was determined by flame atomic absorption spectroscopy. The results indicate that the initial Cr(VI) concentration negatively influenced growth and the specific growth rate but stimulated the metabolic activity of the three strains; resistance to Cr(VI) by these strains was mainly due to reduction of Cr(VI) rather than chromium bioaccumulation. This study showed the potential ability of these strains as tools for bioremediation of Cr(VI) from contaminated sites.


Bioremediation Cr(VI) removal Cr(VI) tolerant yeasts 



This work was supported by grants from CIUNT and CONICET, Argentina. Technical assistance of Mr. Claudio Delfini and FAAS facilities provided by Dr. Orlando Villegas (Universidad Nacional de San Luis, Argentina) are gratefully acknowledged.


  1. APHA (1992) Standard methods for the examination of water and wastewater, 17th edn. American Public Health Association, Washington DCGoogle Scholar
  2. Baldi F, Vaughan A, Olson G (1990) Chromium(VI)-resistant yeast isolated from a sewage treatment plant receiving tannery wastes. Appl Environ Microbiol 56:913–918PubMedGoogle Scholar
  3. Balsalobre L, Silóniz M, Valderrama M, Benito T, Larrea M, Peinado J (2003) Occurrence of yeasts in municipal wastes and their behaviors in presence o cadmium copper and zinc. J Basic Microbiol 43:185–196PubMedCrossRefGoogle Scholar
  4. Barceloux DG (1999) Chromium. Clin Toxicol 37:173–194CrossRefGoogle Scholar
  5. Bewley RJ (1980) Effects of heavy metal pollution on oak leaf microorganisms. Appl Environ Microbiol 40:1053–1059PubMedGoogle Scholar
  6. Biedermann KA, Landolph JR (1990) Role of valence state and solubility of chromium compounds on induction of cytotoxicity, mutagenesis, and anchorage independence in diploid human fibroblasts. Cancer Res 50:7835–7842PubMedGoogle Scholar
  7. Breierová E, Vajczikova I, Sasinková V, Stratilová E, Fišera M, Gregor T, Šajbidor J (2002) Biosorption of cadmium ions by different yeast species. Z Naturforsch 57c:634–639Google Scholar
  8. Cabrera G, Viera M, Gómez J, Cantero D, Donati E (2007) Bacterial removal of chromium (VI) and (III) in continuous system. Biodegradation 18:505–513Google Scholar
  9. Cervantes C, Campos-García J, Devars S, Gutiérrez-Corona F, Loza-Tavera H, Torres-Guzmán JC, Moreno-Sáncez R (2001) Interactions of chromium with microorganisms and plants. FEMS Microbiol Rev 25:335–347PubMedCrossRefGoogle Scholar
  10. Debski B, Zalewski W, Gralak MA, Kosla T (2004) Chromium-yeast supplementation of chicken broilers in an industrial farming system. J Trace Elem Med Biol 18:47–51PubMedCrossRefGoogle Scholar
  11. Dönmez G, Aksu Z (2001) Bioaccumulation of copper(II) and nickel(II) by the non-adapted and adapted growing Candida sp. Water Res 35:1425–1434PubMedCrossRefGoogle Scholar
  12. Dursun AY, Uslu G, Cuci Y, Aksu Z (2003) Bioaccumulation of copper(II), lead(II) and chromium(VI) by growing Aspergillus niger. Process Biochem 38:1647–1651CrossRefGoogle Scholar
  13. Gulan Zetic V, Stehlik Tomas V, Grba S, Lutilsky L, Kozlek D (2001) Chromium uptake by Saccharomyces cerevisiae and isolation of glucose tolerance factor from yeast biomass. J Biosci 23:217–223CrossRefGoogle Scholar
  14. Iyer A, Mody K, Jha B (2004) Accumulation of hexavalen chromium by an exopolysaccharide producing marine Enterobacter cloaceae. Mar Pollut Bull 49:974–977PubMedCrossRefGoogle Scholar
  15. Iyer A, Mody K, Jha B (2005) Biosorption of heavy metals by a marine bacterium. Mar Pollut Bull 50:340–343PubMedCrossRefGoogle Scholar
  16. Kaszycki P, Fedorovych D, Ksheminiska H, Babyak L, Wójcik D, Koloczek H (2004) Chromium accumulation by living yeast at various environmental conditions. Microbiol Res 159:11–17PubMedCrossRefGoogle Scholar
  17. Krauter P, Martinelli R, Williams K, Martins S (1996) Removal of Cr(VI) from ground water by Saccharomyces cerevisiae. Biodegradation 7:277–286PubMedCrossRefGoogle Scholar
  18. Ksheminska H, Jaglarz A, Fedorovych D, Babyak L, Yanovych D, Kaszycki P, Koloczek H (2003) Bioremediation of chromium by the yeast Pichia guilliermondii: toxicity and accumulation of Cr(III) and Cr(VI) and the influence of riboflavin on Cr tolerance. Microbiol Res 158:59–67PubMedCrossRefGoogle Scholar
  19. Ksheminska H, Fedorovych D, Babyak L, Yanovych D, Kaszycki P, Koloczek H (2005) Chromium(III) and (VI) tolerance and bioaccumulation in yeast: a survey of cellular chromium content in selected strains of representative genera. Process Biochem 40:1565–1572CrossRefGoogle Scholar
  20. Kurtzman CP, Fell JW (1998) Definition, classification and nomenclature of the yeasts. In: Kurtzman CP, Fell JW (eds) The yeasts, a taxonomic study, 4th edn. Elsevier Science BV, Amsterdam, pp 2–5Google Scholar
  21. Kurtzman CP, Robnett C (1998) Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. Antonie Van Leeuwenhoek 73:331–371PubMedCrossRefGoogle Scholar
  22. Laxman R, More S (2002) Reduction of hexavalent chromium by Streptomyces griseus. Miner Eng 15:831–837CrossRefGoogle Scholar
  23. Liu Y, Xu W, Zeng G, Li X, Goa H (2006) Cr(CI) reduction by Bacillus sp. Isolated from chromium landfill. Process Biochem 41:1981–1986CrossRefGoogle Scholar
  24. López-Archilla A, González A, Terron M, Amils R (2004) Ecological study of the fungal populations of the acidic Tinto River in southwestern Spain. Can J Microbiol 50:923–934PubMedCrossRefGoogle Scholar
  25. Mikes J, Siglova M, Cejkova A, Masak J, Jirku V (2005) The influence of heavy metals on the production of extracellular polymer substance in the processes of heavy metal ions elimination. Water Sci Technol 52:151–156PubMedGoogle Scholar
  26. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428CrossRefGoogle Scholar
  27. Muter O, Lubinya I, Millers D, Grigorjeva L, Ventinya E, Rapoport A (2002) Cr(VI) sorption by intact and dehydrated Candida utilis cells in the presence of other metals. Process Biochem 38:123–131CrossRefGoogle Scholar
  28. Orozco AM, Contreras EM, Bertola N, Zrotzky NE (2007) Hexavalent chromium removal using arobic activated sludge batch systems added with powdered activated carbon. Water SA 33:239–244Google Scholar
  29. Palmer CH, Wittbrodt PR (1991) Processes affecting the remediation of chromium-contaminated sites. Environ Health Perspect 92:25–40PubMedCrossRefGoogle Scholar
  30. Pas M, Milacic R, Draslar K, Pollak N, Raspor P (2004) Uptake of chromium(III) and chromium(VI) compounds in the yeast cell structure. Biometals 17:25–33PubMedCrossRefGoogle Scholar
  31. Pepi C, Baldi F (1992) Modulation of chromium(VI) toxicity by organic and inorganic sulfur species in yeasts form industrial wastes. Biometals 5:179–185PubMedCrossRefGoogle Scholar
  32. Plaper A, Jenko-Brinovec S, Premzl A, Kos J, Raspor P (2002) Genotoxicity of trivalent chromium in bacterial cells. Possible effects on DNA topology. Chem Res Toxicol 15:943–949PubMedCrossRefGoogle Scholar
  33. Ramírez-Ramírez R, Calvo-Méndez C, Ávila-Rodríguez M, Lappe P, Ulloa M, Vázquez-Juárez R, Gutiérrez-Corono F (2004) Cr(VI) reduction in a chromate-resistant strain of Candida maltosa isolated from the leather industry. Antonie Van Leeuwenhoek 85:63–68PubMedCrossRefGoogle Scholar
  34. Raspor P, Batic M, Jamnik P, Josic D, Milacic R, Pas M, Recek M, Rezic-Dereani V, Skrt M (2000) The influence of chromium compounds on yeast physiology. Acta Microbiol Immunol Hung 47:143–173PubMedCrossRefGoogle Scholar
  35. Ross IS, Parkin MJ (1989) Uptake of copper by Candida utilis. Mycol Res 93:33–37CrossRefGoogle Scholar
  36. Sambrook J, Maniatis T, Fritsch EF (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory PressGoogle Scholar
  37. Sampaio JP, Gadanho M, Santos S, Duarte FL, Pais C, Fonseca A, Fell JW (2001) Polyphasic taxonomy of the basidiomycetous yeast genus Rhodosporidium: Rhodosporidium kratochvilovae and related anamorphic species. Int J Syst Evol Microbiol 51:687–697PubMedGoogle Scholar
  38. Santos V, Linardi V (2001) Phenol degradation by yeasts isolated from industrial effluents. J Gen Appl Microbiol 47:213–221PubMedCrossRefGoogle Scholar
  39. Silóniz M, Balsolobre L, Alba C, Valderrama M, Peinado J (2002) Feasibility of copper uptake by the yeast Pichia guilliermondii isolated form sewage sludge. Res Microbiol 153:173–180PubMedCrossRefGoogle Scholar
  40. Srinath T, Verma T, Ramteke PW, Garg SK (2002) Chromium (VI) biosorption and bioaccumulation by chromate resistant bacteria. Chemosphere 48:427–435PubMedCrossRefGoogle Scholar
  41. Srivastava S, Thakur IS (2006) Isolation and process parameter optimization of Aspergillus sp. for removal of chromium from tannery effluent. Bioresour Technol 97:1167–1173PubMedCrossRefGoogle Scholar
  42. Tang YJ, Laidlaw D, Gani K, Keasling JD (2006) Evaluation of the effects of various culture conditions on Cr(VI) reduction by Shewanella oneidensis MR-1 in a novel high-throughput mini-bioreactor. Biotechnol Bioeng 95:176–184PubMedCrossRefGoogle Scholar
  43. Villegas L, Amoroso MJ, Figueroa LIC (2004) Selection of tolerant heavy metal yeasts from different polluted sites. In: Spencer JFT, Spencer ALR (eds) Methods in biotechnology, vol 16: environmental biology, methods in protocols. Humana Press Inc., Totowa, pp 249–256Google Scholar
  44. Villegas L, Amoroso MJ, Figueroa LIC (2005) Copper tolerant yeasts isolated from polluted area of Argentina. J Basic Microbiol 45:383–391CrossRefGoogle Scholar
  45. Wang Y, Xiao C (1995) Factors affecting hexavalent chromium reduction in pure cultures of bacteria. Water Res 29:2467–2474CrossRefGoogle Scholar
  46. Weber E, Görke C, Begerow D (2002) The Lecythophora-Coniochaeta complex II. Molecular studies based on sequences of the large subunit of ribosomal DNA. Nova Hedwigia 74:187–200CrossRefGoogle Scholar
  47. Yarrow D (1998) Methods for the isolation, maintenance and identification of yeasts. In: Kurtzman CP, Fell JW (eds) The yeasts, a taxonomic study, 4th edn. Elsevier Science BV, Amsterdam, pp 77–100Google Scholar
  48. Zayed AM, Terry N (2003) Chromium in the environment: factors affecting biological remediation. Plant Soil 249:139–156CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2008

Authors and Affiliations

  1. 1.PROIMI – Biotecnología – CONICETTucumanArgentina
  2. 2.Microbiología General, Facultad de Bioquímica, Química y FarmaciaUniversidad Nacional de TucumánTucumanArgentina
  3. 3.Microbiología Superior, Facultad de Bioquímica, Química y FarmaciaUniversidad Nacional de TucumánTucumanArgentina

Personalised recommendations