The kinetic reduction of Cr(VI) by yeast Saccharomyces cerevisiae, Phaffia rhodozyma and their protoplasts
Chromium in the sixth oxidation state may easily penetrate cellular membranes via non-specific sulfate transporters due to its tetrahedral symmetry (high similarity to SO42- and HPO42-). This feature makes chromium a toxic and hazardous pollutant responsible for the deterioration of midland water quality. The aim of the study was to evaluate the capacity of two yeast species - Saccharomyces cerevisiae and Phaffia rhodozyma - and their protoplasts to reduce Cr(VI) to lower oxidation states. The study also deals with the behavior of the yeasts upon the presence of elevated sulfate ions as a competitive inhibitor of chromate transport by the sulfate transporters. The chromate-reducing activities were monitored by determination of Cr(V) free radical form with the use of L-band (1.2 GHz) EPR (electron paramagnetic resonance) spectroscopy. It was observed that both of the studied yeast strains exhibited the ability to reduce Cr(VI) applied at 4 mM. The cells of P. rhodozyma showed about 3.5 times higher reduction than S. cerevisiae. The reduction efficiency was significantly improved when the protoplasts of both strains were used and reached 100% in the first 10 minutes of the reduction process which suggests that the cellular wall may have a notable influence on the uptake and/or inhibition of chromium reduction process. The reduction effect of P. rhodozyma cells and protoplasts may be associated with the more sufficient production of metabolites (such as glutathione and cysteine), which may also be responsible for the increased tolerance of the strain towards high concentrations of toxic chromium.
- Abbas Ch, Sibirny (2011) Genetic control of biosynthesis and transport of riboflavin and flavin nucleotides and construction of robust biotechnological producers. Microbiol Mol Biol Rev 75: 321-360.
- Appenroth D, Rost M, Friedrich M, Schulz O (1996) Contribution to the mechanism of chromate nephrotoxicity in developing rats: EPR investigations. Arch Toxicol 71: 25-30.
- Babyak L, Kseminskaya G, Gonchar M, Yanovich D, Fedorovich D. (2005) Selection and properties of mutant yeast Pichia guilliermondii strais resistant to chromium (VI) Appl Biochem Microbiol 41: 177-181.
- Barceloux DG (1999) Chromium. J Toxicol Clin Toxicol 37: 173-194.
- Cervantes C, Campos-Garcia J, Devars S, Gutierrez-Corona F, Loza-Tavera H, Torres-Guzman JC, Moreno-Sanchez R (2001) Interactions of chromium with microorganisms and plants. FEMS Microbiol 25: 335-347.
- Cherest H, Davidian JC, Thomas D, Benes V, Ansorge W, Surdin-Kerjan Y (1997) Molecular characterization of two high affinity sulfate transporters in Saccharomyces cerevisiae. Genetics 145: 627-635.
- Cieślak-Golonka (1996) Toxic and mutagenic effects of chromium (VI). A Review. Polyhedron 21: 3667-3689.
- Fedorovych D, Kszeminska H, Babjak D, Kaszycki P, Koloczek H (2001) Hexavalent chromium stimulation of riboflavin synthesis in flavinogenic yeast. BioMetals 14: 23-31.
- Headlam HA, Lay PA (2001) EPR spectroscopic studies of the reduction of chromium (VI) by methanol in presence of peptides. Formation of long-lived chromium (V) peptide complexes. Inorg Chem 40: 78-86.
- Horvath E, Papp G, Gazdag Z, Belagyi J, Blasko A, Deli J, Vagvolgyi Cs, Pesti M (2011) Characterization of stress processes of Phaffia rhodozyma stress-resistant mutant. Acta Biol Hung 62: 204-210.
- Jamnik P, Raspor P (2003) Stress response of yeast Candida intermedia to Cr(VI). Biochem Mol Toxicol 17: 316-323.
- Kaszycki P, Gabryś H, Appenroth K.J, Jaglarz A, Sedziwy S, Walczak T, Koloczek H (2005) Exogenously applied sulphate as a tool to investigate transport and reduction ofchromate in the duckweed Spirodela polyrhiza. Plant Cell Environ 28: 260-268.
- 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: 1565-1572.
- Ksheminska HP, Honchar T, Gayda G, Gonchar M (2006) Extra-cellular chromate-reducing activity of the yeast cultures. CEJB 1: 137-149.
- Ksheminska H, Honchar T, Usatenko Y, Gayda G, Gonchar M (2010) The chromate resistance phenotype of some yeast mutants correlates with a lower level of Cr(V)-species generated in the extra-cellular medium. Biometals 23: 633-642.
- Nechay HI, Ksheminska P, Kolisnyk HV, Grządka M, Gonchar MV (2009) Reduction of chromate and carotene-synthesizing activity of selenite-resistant mutants of the yeast Xanthophyllomycesdendrorhous (Phaffia rhodozyma). Biopolymers and Cell 25: 633-642.
- Pas M, Milacic R, Draslar K, Pollak N, Raspor P (2004) Uptake of chromium(III) and chromium(VI) compunds in the yeast cell structure. BioMetals 17: 25-33.
- Pepi M, Baldi F (1992) Modulation of chromium(VI) toxicity by organic and inorganic sulfur species in yeasts from industrial wastes. Biometals 5: 179-185.
- Pereira Y, Lagniel G, Godat E, Baudouin-Cornu P, Junot Ch, Labarre J (2008) Chromate causes sulfur starvation in yeast. Toxicol Sci 106: 400-412.
- Pesti M, Gazdag Z, Emri T, Farkas N, Koosz Z, Belagyi J, Pocsi I (2002) Chromate sensitivity in fission yeast is caused by increased glutathione reductase activity and peroxide overproduction. J Basic Microbiol 42: 408-419.
- Sami M, Ikeda M, Yabuuchi S (1994) Evaluation of the alkaline methylene bluestaining method for yeast activity determination. J Ferment Bioengin 78: 212-216.
- Shi X, Dong Z, Dalal NS, Gannet PM (1994) Chromate-mediated free radical generation from cysteine, penicillamine, hydrogen peroxide and lipid hydroperoxides. Biochim Biophys Acta 1226: 65-72.
- Smutok O, Broda D, Smutok H, Dmytruk K, Gonchar M (2011) Chromate-reducing activity of Hansenula polymorpha recombinant cells over-producing flavocytochrome b2. Chemosphere 83: 449-454.
- Wysocki R, Tamas MJ (2010) How Saccharomyces cerevisiae cope with toxic metals and metalloids. FEMS Microbiol 34: 925-951.
- Yoshimoto N, Takahashi H, Smith FW, Yamaya T, Saito K (2002) Two distinct high-affinity sulfate transporters with different inducibilities mediate uptake of sulfate in iArabidopsis roots. Plant J 29: 465-473.