BioMetals

, Volume 20, Issue 5, pp 773–780 | Cite as

Characteristics of copper tolerance in Yarrowia lipolytica

  • Hiroyasu Ito
  • Masahiro Inouhe
  • Hiroshi Tohoyama
  • Masanori Joho
Article

Abstract

We discovered that a mutant strain of the dimorphic yeast Yarrowia lipolytica could grow in the yeast form in high concentrations of copper sulfate. The amount of metal accumulated by Y. lipolytica increased with increasing copper concentrations in the medium. Washing with 100 mM EDTA released at least 60% of the total metal from the cells, but about 20–25 μmol/g DW persisted, which represented about 30% of the soluble fraction of cultured cells. The soluble fraction (mainly cytosol) contained only about 10% of the total metal content within cells cultured in medium supplemented with 6 mM copper. We suggest that although a high copper concentration induces an efflux mechanism, the released copper becomes entrapped in the periplasm and in other parts of the cell wall. Washing with EDTA liberated not only copper ions, but also melanin, a brown pigment that can bind metal and which located at the cell wall. These findings indicated that melanin participates in the mechanism of metal accumulation. Culture in medium supplemented with copper obviously enhanced the activities of Cu, Zn-SOD, but not of Mn-SOD.

Keywords

copper tolerance melanin superoxide dismutase (SOD) activity Yarrowia lipolytica 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Andreishcheva EN, Isakova EP, Sidorov NN, et al. 1999 Adaptation to salt stress in a salt-tolerant strain of the yeast Yarrowia lipolytica. Biochemistry (Moscow) 64:1061–1067Google Scholar
  2. Antonucci S, Bravi M, Bubbico R, Di Michele A, Verdone N. 2001 Selectivity in citric acid production by Yarrowia lipolytica Enzyme Microb Technol 28:189–195PubMedCrossRefGoogle Scholar
  3. Bartnicki-Garcia S, Nickerson WJ. 1962 Isolation, composition, and structure of cell walls of filamentous and yeast-like forms of Mucor rouxii Biochem Biophys Acta 58:102–119PubMedCrossRefGoogle Scholar
  4. Barth G, Gaillardin C. 1997 Physiology and genetics of the dimorphic fungus Yarrowia lipolytica. FEMS Microbiol Rev 19:219–237PubMedCrossRefGoogle Scholar
  5. Borghouts C, Osiewacz HD. 1998 GRISEA, a copper-modulated transcription factor from Podospora anserina involved in senescence and morphogenesis, is an ortholog of MAC1 in Saccharomyces cerevisiae. Mol Gen Genet 260:492–502PubMedCrossRefGoogle Scholar
  6. Brady D, Duncan JR. 1994 Binding of heavy metals by the cell walls of Saccharomyces cerevisiae. Enzyme Microb Technol 16:633–638CrossRefGoogle Scholar
  7. Brady D, Stoll AD, Starke L, Duncan JR. 1994 Chemical and enzymatic extraction of heavy metal binding polymers from isolated cell walls of Saccharomyces cerevisiae Biotechnol Bioeng 44:297–302CrossRefPubMedGoogle Scholar
  8. Butinar L, Santos S, Spencer-Martins I, Oren A, Gunde-Cimerman N. 2005 Yeast diversity in hypersaline habitats. FEMS Microbiol Lett 244:229–234PubMedCrossRefGoogle Scholar
  9. Butler MJ, Lazarovits G, Higgins VJ, Lachance MA. 1989 Identification of a black yeast isolated from oak bark as belonging to the genus Phaeococcomyces sp. Analysis of melanin produced by the yeast. Can J Microbiol 35:728–734CrossRefGoogle Scholar
  10. Butler MJ, Day AW. 1998 Fungal melanins: a review Can J Microbiol 44:1115–1136CrossRefGoogle Scholar
  11. Carreira A, Loureiro V. 1998 A differential medium to detect Yarrowia lipolytica within 24 hours. J Food Mycol 1:3–12Google Scholar
  12. Carreira A, Ferreira LM, Loureiro V. 2001 Brown pigments produced by Yarrowia lipolytica result from extracellular accumulation of homogentisic acid. Appl Environ Microbiol 67:3463–3468PubMedCrossRefGoogle Scholar
  13. Dancis A, Haile D, Yuan DS, Klausner RD. 1994 The Saccharomyces cerevisiae copper transport protein (Ctr1p). Biochemical characterization, regulation by copper, and physiological role in copper uptake. J Biol Chem 269:25660–25667PubMedGoogle Scholar
  14. Elstner EF, Heupel A. 1976 Inhibition of nitrite formation from hydroxylammonium chloride: a simple assay for superoxide dismutase. Anal Biochem 70:616–620PubMedCrossRefGoogle Scholar
  15. Elorza MV, Marcilla A, Sentandreu R. 1988 Wall mannoproteins of the yeast and mycelial cells of Candida albicans: nature of the glycosidic bonds and polydispersity of their mannan moieties. J Gen Microbiol 134:2393–2403PubMedGoogle Scholar
  16. Fickers P, Nicaud JM, Gaillardin C, Destain J, Thonart P. 2004 Carbon and nitrogen sources modulate lipase production in the yeast Yarrowia lipolytica. J Appl Microbiol 96:742–749PubMedCrossRefGoogle Scholar
  17. Fogarty RV, Tobin JM. 1996 Fungal melanins and their interactions with metals. Enzyme Microb Technol 19:311–317PubMedCrossRefGoogle Scholar
  18. Gadd GM, Griffiths AJ. 1980 Effect of copper on morphology of Aureobasidium pullulans Trans Br Mycol Soc 74:387–392CrossRefGoogle Scholar
  19. Gadd GM, de Rome L. 1988 Biosorption of copper by fungal melanin. Appl Microbiol Biotechnol 29:610–617CrossRefGoogle Scholar
  20. Gadd GM, Mowll JL. 1985 Copper uptake by yeast-like cells, hyphae, and chlamydospores of Aureobasidium pullulans Exp Mycol 9:230–240CrossRefGoogle Scholar
  21. Hurtado CAR, Rachubinski RA. 1999 MHY1 encodes a C2H2-type zinc finger protein that promotes dimorphic transition in the yeast Yarrowia lipolytica. J Bacteriol 181:3051–3057PubMedGoogle Scholar
  22. Jensen LT, Winge DR. 1998 Identification of a copper-induced intramolecular interaction in the transcription factor Mac1 from Saccharomyces cerevisiae. EMBO J 17:5400–5408PubMedCrossRefGoogle Scholar
  23. Kapoor A, Viraraghavan T. 1997 Heavy metal biosorption sites in Aspergillus niger. Bioresour Technol 61:221–227CrossRefGoogle Scholar
  24. Kierans M, Staines AM, Bennett H, Gadd GM. 1991 Silver tolerance and accumulation in yeasts. Biol Met 4:100–106PubMedCrossRefGoogle Scholar
  25. Lapinskas PJ, Cunningham KW, Liu XF, Fink GR, Culotta VC. 1995 Mutations in PMR1 suppress oxidative damage in yeast cells lacking superoxide dismutase. Mol Cell Biol 15:1382–1388PubMedGoogle Scholar
  26. Lee JK, Kim JM, Kim SW, Nam DH, Yong CS, Huh K. 1996 Effect of copper ion on oxygen damage in superoxide dismutase-deficient Saccharomyces cerevisiae. Arch Pharm Res 19:178–182CrossRefGoogle Scholar
  27. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951 Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  28. Manzano M, Cocolin L, Citterio B, et al. 2000 Biochemical responses in a Candida famata strain adapted to high copper concentrations. BioMetals 13:251–259PubMedCrossRefGoogle Scholar
  29. Margesin R, Schinner F. 1997 Effect of temperature on oil degradation by a psychrotrophic yeast in liquid culture and in soil. FEMS Microbiol Ecol 24:243–249CrossRefGoogle Scholar
  30. Melo RGM, Leitão AC, Pádula M. 2004 Role of OGG1 and NTG2 in the repair of oxidative DNA damage and mutagenesis induced by hydrogen peroxide in Saccharomyces cerevisiae: relationships with transition metals iron and copper. Yeast 21:991–1003PubMedCrossRefGoogle Scholar
  31. Mowll JL, Gadd GM. 1984 Cadmium uptake by Aureobasidium pullulans. J Gen Microbiol 130:279–284Google Scholar
  32. Naiki N. 1980 Role of superoxide dismutase in a copper-resistant strain of yeast. Plant Cell Physiol 21:775–783Google Scholar
  33. Ramezani Rad M, Kirchrath L, Hollenberg CP. 1994 A putative P-type Cu2+- transporting ATPase gene on chromosome II of Saccharomyces cerevisiae. Yeast 10:1217–1225CrossRefGoogle Scholar
  34. Riggle PJ, Kumamoto CA. 2000 Role of a Candida albicans P1-type ATPase in resistance to copper and silver ion toxicity. J Bacteriol 182:4899–4905PubMedCrossRefGoogle Scholar
  35. Ruiz-Herrera J, Sentandreu R. 2002 Different effectors of dimorphism in Yarrowia lipolytica. Arch Microbiol 178:477–483PubMedCrossRefGoogle Scholar
  36. Samarelli J Jr, Campbell WH. 1983 Heavy metal inactivation and chelators stimulation of higher plant nitrate reductase. Biochim Biophys Acta 742:435–445Google Scholar
  37. Sarais I, Manzano M, de Bertoldi M, et al. 1994 Adaptation of a Saccharomyces cerevisiae strain to high copper concentrations. BioMetals 7:221–226PubMedCrossRefGoogle Scholar
  38. Shanmuganathan A, Avery SV, Willetts SA, Houghton JE. 2004 Copper-induced oxidative stress in Saccharomyces cerevisiae targets enzymes of the glycolytic pathway. FEBS Lett 556:253–259PubMedCrossRefGoogle Scholar
  39. Shi X, Stoj C, Romeo A, Kosman DJ, Zhu Z. 2003 Fre1p Cu2+ reduction and Fet3p Cu1+ oxidation modulate copper toxicity in Saccharomyces cerevisiae. J Biol Chem 278:50309–50315PubMedCrossRefGoogle Scholar
  40. Shiraishi E, Inouhe M, Joho M, Tohoyama H. 2000 The cadmium-resistant gene, CAD2, which is a mutated putative copper-transporter gene (PCA1), controls the intracellular cadmium-level in the yeast S. cerevisiae. Curr Genet 37:79–86PubMedCrossRefGoogle Scholar
  41. Soares EV, Hebbelinck K, Soares HMVM. 2003 Toxic effects caused by heavy metals in the yeast Saccharomyces cerevisiae: a comparative study. Can J Microbiol 49:336–343PubMedCrossRefGoogle Scholar
  42. Strouhal M, Kizek R, Vacek J, Trnková L, Némec M. 2003 Electrochemical study of heavy metals and metallothionein in yeast Yarrowia lipolytica. Bioelectrochemistry 60:29–36PubMedCrossRefGoogle Scholar
  43. Suresh K, Subramanyam C. 1998 Polyphenols are involved in copper binding to cell walls of Neurospora crassa. J Inorg Biochem 69:209–215CrossRefGoogle Scholar
  44. Szabo R, Stofaniková V. 2002 Presence of organic sources of nitrogen is critical for filament formation and pH-dependent morphogenesis in Yarrowia lipolytica. FEMS Microbiol Lett 206:45–50PubMedCrossRefGoogle Scholar
  45. Venkateswerlu G, Stotzky G. 1986 Copper and cobalt alter the cell wall composition of Cunninghamella blakesleeana. Can J Microbiol 32:654–662PubMedCrossRefGoogle Scholar
  46. Yamaguchi-Iwai Y, Serpe M, Haile D, et al. 1997 Homeostatic regulation of copper uptake in yeast via direct binding of MAC1 protein to upstream regulatory sequences of FRE1 and CTR1. J Biol Chem 272:17711–17718PubMedCrossRefGoogle Scholar
  47. Yu W, Farrell RA, Stillman DJ, Winge DR. 1996 Identification of SLF1 as a new copper homeostasis gene involved in copper sulfide mineralization in Saccharomyces cerevisae. Mol Cell Biol 16:2464–2472PubMedGoogle Scholar
  48. Zinjarde SS, Pant AA. 2002a Hydrocarbon degraders from tropical marine environments. Mar Pollut Bull 44:118–121CrossRefGoogle Scholar
  49. Zinjarde SS, Pant AA. 2002b Emulsifier from a tropical marine yeast, Yarrowa lipolytica NCIM 3589. J Basic Microbiol 1:67–73CrossRefGoogle Scholar
  50. Zvyagilskaya R, Andreishcheva E, Soares MIM, et al. 2001 Isolation and characterization of a novel leaf-inhabiting osmo-, salt-, and alkaline-tolerant Yarrowia lipolytica yeast strain. J Basic Microbiol 41:289–303PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Hiroyasu Ito
    • 1
  • Masahiro Inouhe
    • 1
  • Hiroshi Tohoyama
    • 1
  • Masanori Joho
    • 1
  1. 1.Department of Biology, Faculty of ScienceEhime UniversityMatsuyamaJapan

Personalised recommendations