Efficient removal of hexavalent chromium by a tolerant Streptomyces sp. affected by the toxic effect of metal exposure

AIMS: To isolate and analyse chromium-resistant micro-organisms suitable for bioremediation. METHODS AND RESULTS: Strain CG252, with a minimal inhibitory concentration of 500 microg ml(-1), was isolated from contaminated soils and identified as a Streptomyces sp. by 16S rDNA sequence analysis. Assays carried out at various Cr(VI) concentrations indicated that chromium removal was more efficient at lower concentrations and that this activity resulted in accumulation of Cr(III). Atomic adsorption analysis indicated that the chromium removed was not associated with cell mass and activity assays showed that the capacity to reduce Cr(VI) was most probably due to a soluble cytosolic enzyme. Cells grown as biofilms showed enhanced removal of Cr(VI) with respect to planktonic cells, while analysis of growth and colony morphology indicated that Cr(VI) had a toxic effect on this strain. CONCLUSIONS: Streptomyces sp. CG252 tolerated heavy metals and elevated levels of chromium, despite its negative effect on growth and development, and was efficient at removing Cr(VI) by promoting reduction to Cr(III). SIGNIFICANCE AND IMPACT OF THE STUDY: Strain CG252's capacity to tolerate heavy metals and to reduce Cr(VI) to the less toxic Cr(III), especially when forming biofilms, makes it a promising candidate for detoxification of sites containing this heavy metal.     

Reductive Immobilization of Chromium in Soils Containing Heterogeneous Fe-Bearing Minerals

Cr(VI) immobilization in systems containing Fe-bearing soil minerals was studied in batch and column systems. Batch experiments showed that water chemistry such as solution pH and Cr(VI) concentration had a pronounced impact on Cr(VI) removal by Fe-bearing soil minerals. Acidic conditions were observed to be more favorable for enhanced Cr(VI) removal. The dependence of Cr(VI) removal on Cr(VI) concentration indicated that there were limited numbers of surface sites on Fe-bearing minerals responsible for Cr(VI) removal. A complexing agent, citrate, significantly enhanced both Cr(VI) removal and total Fe-dissolution from the mineral surfaces relative to non-citrate containing systems, and the iron dissolved from the mineral surfaces was in Fe(III) oxidation form, implying that Cr(VI) removal occurred mainly on mineral surfaces, and the surface Fe(II) sites played an active role in Cr(VI) reduction. The results from column experiments showed that the accumulation of surface precipitates resulted in clogging of pore spaces, thereby creating preferential flow paths within the column. However, the addition of citrate significantly prevented the accumulation of surface precipitates due to the formation of highly soluble Fe–citrate complexes. SEM images revealed that the precipitates accumulated in the column had sponge-like shapes. The energy-dispersive spectroscopy analysis provided further evidence that the surface precipitates formed also contained Cr species as well as Fe. Overall it is clear that Fe-bearing minerals may serve as an effective reducing agent for in-situ reductive immobilization of hexavalent chromium in subsurface systems.     

Hexavalent chromium reduction by bacterial consortia and pure strains from an alkaline industrial effluent

Aims: To characterize the bacterial consortia and isolates selected for their role in hexavalent chromium removal by adsorption and reduction. Methods and Results: Bacterial consortia from industrial wastes revealed significant Cr(VI) removal after 15 days when incubated in medium M9 at pH 6·5 and 8·0. The results suggested chromium reduction. The bacterial consortia diversity (T‐RFLP based on 16S rRNA gene) indicated a highest number of operational taxonomic units in an alkaline carbonate medium mimicking in situ conditions. However, incubations under such conditions revealed low Cr(VI) removal. Genomic libraries were obtained for the consortia exhibiting optimal Cr(VI) removal (M9 medium at pH 6·5 and 8·0). They revealed the dominance of 16S rRNA gene sequences related to the genera Pseudomonas/Stenotrophomonas or Enterobacter/Halomonas, respectively. Isolates related to Pseudomonas fluorescens and Enterobacter aerogenes were efficient in Cr(VI) reduction and adsorption to the biomass. Conclusions: Cr(VI) reduction was better at neutral pH rather than under in situ conditions (alkaline pH with carbonate). Isolated strains exhibited significant capacity for Cr(VI) reduction and adsorption. Significance and Impact of Study: Bacterial communities from chromium‐contaminated industrial wastes as well as isolates were able to remove Cr(VI). The results suggest a good potential for bioremediation of industrial wastes when optimal conditions are applied.     

Biosorption of trivalent and hexavalent chromium from aqueous systems using shelled Moringa oleifera seeds

Abstract

The present study explores the sorption properties of shelled Moringa oleifera seeds (SMOS) for removal of two environmentally important oxidation states of chromium (trivalent and hexavalent) from an aqueous system on the laboratory scale. Sorption studies reveal the optimum conditions for the removal of 81.02%; Cr (III) and 88.15% Cr (VI) as follows: biomass dosage (4.0 g), metal concentration [25mg/L for Cr (III); 50mg/L for Cr (VI)], contact time (40 minutes) at pH 6.5 and 2.5 respectively. The adsorption data were found to fit well both the Freundlich and Langmuir isotherms. Characterization of the seed powder by FTIR showed the clear presence of amino acid moieties having both positively charged amino and negatively charged carboxylic groups and confirmed that biosorption involves amino acid-chromium interactions. SEM studies of native and exhausted [Cr(III) and Cr(VI)] treated SMOS revealed large spherical clusters having a pore area of 8.66 µm² in the case of native SMOS while dense agglomerated etched dendrite type morphology have a pore area of 0.80 µm² in Cr (III) and 0.78 µm² in Cr (VI) treated SMOS The spent biosorbent was regenerated and found to be effectively reusable for four cycles.     

Optimization of Chromium(VI) Detoxification by Pseudomonas aeruginosa and Its Application for Treatment of Industrial Waste and Contaminated Soil

The reduction of Cr(VI) to Cr(III) is a potential detoxification process. In this study, seven Pseudomonas spp. were isolated and screened for chromium reduction. Isolate P4 was able to grow in the presence of 8000 μM chromium, in spite of the fact that the isolate was not previously exposed to any metal stress. Isolate P4 was identified as Pseudomonas aeruginosa strain SRD chr3 by 16S rDNA sequence analysis. Shake flask study showed 78% reduction of 1000 μM Cr(VI) after 6 h of incubation. The optimum pH for chromium reduction by the isolate was between 6 and 8. Isolate Pseudomonas aeruginosa gave 50–80% Cr(VI) reduction even in the presence of 100 mM of Cu, Mn, Ni, and Zn and 300–800 mM NaCl in 24 h, compared with the absence of any of these metals. In a 5-L reactor, the isolate showed 84.85% reduction of Cr(VI) even at the 70th cycle, with a hydraulic retention time of 24 h from the effluent of a hard chrome plating (electroplating) industry, which contained 2100 mg L^?1 hexavalent chromium. The chromate-amended soil inoculated with the isolate showed 2800 μM chromium removal from 4000 μM Cr(VI) kg^?1 of soil, which corresponds to 70% removal. The isolate had the ability to degrade stimulated waste containing 10,000 μM chromium.     

Removal of Chromium (VI) from Aqueous Solution Using Mycelial Pellets of Penicillium simplicissimum Impregnated with Powdered Biochar

The mycelia pellets of Penicillium simplicissimum impregnated with powdered biochar (MPPSIPB) were synthesized and applied to remove chromium (VI) from aqueous solution. The effects of pH, MPPSIPB dosage, initial Cr(VI) concentration, and contact time were investigated via batch experiments. Results indicated that the percentage removal of Cr(VI) was significantly dependent on the pH of the solution. Ten grams mycelial pellets and 0.2 g powdered biochar could form the most stable pellets. The maximum value of biosorption of Cr(VI) was 28.0 mg/g. Scanning electron microscopy (SEM) analysis showed that the mycelia pellets of Penicillium simplicissimum had abundant filamentous network, which entrapped powdered biochar firmly. Fourier transform infrared (FTIR) analysis suggested that O?H, N?H, C?H, C?O, and C?OH groups from MPPSIPB were involved in chromium binding and the subsequent reduction. Kinetic studies indicated that the pseudo-second-order equation fit best for Cr(VI) removal from aqueous solution. Freundlich isotherm was found to apply better for the adsorption equilibrium data with respect to the Langmuir isotherm. Furthermore, MPPSIPB can be separated from aqueous solution completely by filtration. Both experimental study and modeling results indicated that MPPSIPB exhibited remarkable affinity for chromate and had a potential application in Cr(VI) removal from water.     

Biosorption of chromium (Cr(III)/Cr(VI)) on the residual microalga Nannochloris oculata after lipid extraction for biodiesel production

In order to increase the economic feasibility of biodiesel production from microalgae, the residual biomass after biodiesel production can be utilized as biosorbent for heavy metal removal. In this study, biosorption of chromium by residual Nannochloris oculata after lipid extraction was investigated. Increased surface area of N. oculata was observed after lipid extraction. Cr(III) removal increased as the pH increased from 2 to 6, while Cr(VI) removal was highest at pH 2 and it decreased with the increase in pH. Cr(VI) was reduced to Cr(III) in the presence of biomass under acidic conditions; X-ray photoelectron spectroscopy revealed that the converted Cr(III) was bound to the biomass. Chromium removal was significantly enhanced at high chromium concentrations, which indicates that surface reactions may occur at high chromium/biomass ratios. FTIR study indicated that phosphate and carboxyl functional groups of the biomass were mainly responsible for chromium binding.     

A low-GC Gram-positive Thermoanaerobacter-like bacterium isolated from an Indian hot spring contains Cr(VI) reduction activity both in the membrane and cytoplasm

Aim:  Characterization of an anaerobic thermophilic bacterium and subcellular localization of its Cr(VI)-reducing activity for potential bioremediation applications.
Methods and Results:  16S rRNA gene sequence-based analyses of bacterial strains isolated from sediment samples of a Bakreshwar (India) hot spring, enriched anaerobically in iron-reducing medium, found them to be 86–96% similar to reported Thermoanaerobacter strains. The most efficient iron reducer among these, BSB-33, could also reduce Cr(VI) at an optimum temperature of 60°C and pH 6·5. Filtered culture medium could reduce Cr(VI) but not Fe(III). Cell-free extracts reduced Cr(VI) inefficiently under aerobic conditions but efficiently anaerobically. Fractionation of the cell-free extracts showed that chromium reduction activity was present in both the cytoplasm and membrane.
Conclusions:  BSB-33 reduced Fe(III) and Cr(VI) anaerobically at 60°C optimally. After fractionation, the reducing activity of Cr(VI) was found in both cytoplasmic and membrane fractions.
Significance and Impact of the Study:  To the best of our knowledge, this is the first systematic study of anaerobic Cr(VI) reduction by a gram-positive thermophilic micro-organism and, in contrast to our results, none of the earlier reports has mentioned Cr(VI)-reducing activity to be present both in the cytoplasm and membrane of an organism. The strain may offer itself as a potential candidate for bioremediation.     

Effect of organic Cr(III) complexes on chromium speciation

Abstract

Chromium speciation in the presence of organic chromium(III) complexes was investigated using solid-phase extraction. The adsorptions of Cr(VI) and Cr(III) on alumina and pumice powder were studied. Maximum sorption of Cr(VI) was obtained by alumina (90.22%), while Cr(III) was highly adsorbed onto pumice powder (86.65%). This result shows that pumice may be a new and promising adsorbent for Cr(III). The experimental equilibrium data for Cr(VI) adsorption onto alumina and Cr(III) sorption onto pumice were analysed using Langmuir and Freundlich isotherms. The separation and adsorption of Cr(VI), Cr(III) and five organic chromium(III) complexes onto pumice and alumina at different pH values were evaluated. Ethylenediaminetetraacetate (EDTA), oxalate, citrate, glycine, alanine and 8-hydroxyqinoline were used as ligands. Sorption of alanine and ethylenediaminetetraacetate complexes was higher onto alumina than pumice at pH>3. The enhancement of adsorption of chromium(III) complexes onto pumice was achieved by surface modification of pumice using a surfactant, namely hexadecyltrimethylammoniumbromür (HDTMA). The presence of surfactant enhanced the adsorption of Cr(III) citrate, oxalate, glycine and 8-hydroxyquinoline complexes onto pumice. However, the adsorption of EDTA and alanine complexes decreased, with ratio of 13.40% and 4.00% respectively. Here we demonstrate that chromium speciation methods depending on adsorption onto various adsorbents including alumina may lead erroneous results. Analytical measurements were performed by flame AAS, data were obtained by standard addition method.     

Use of Experimental Factorial Design for Optimization of Hexavalent Chromium Removal by a Bacterial Consortium: Soil Microcosm Bioremediation

Hexavalent chromium Cr(VI) is regularly introduced into the environment through diverse anthropogenic activities. It is highly toxic, mutagenic and carcinogenic, and because of its solubility in water, chromate contamination can be difficult to contain. Bacteria can reduce chromate to insoluble and less toxic trivalent chromium Cr(III), and thus increasing attention is paid to chromate bioremediation to reduce its ecotoxicological impacts. In this study, the factorial design 2³ was employed to optimize critical parameters responsible for higher Cr(VI) removal by a bacterial consortium. The factors considered were pH, temperature, and inoculum size at two markedly different levels. All three dependent variables have significant effect on Cr(VI) reduction. Optimal Cr(VI) removal by the bacterial consortium occurred at pH 9, temperature 37°C, and inoculum size OD = 3. Analysis of variance (ANOVA) showed a high coefficient of determination (R²) value of 0.984, thus ensuring a satisfactory adjustment of the second-order regression model with the experimental data. In addition, the effect of bioaugmentation of Cr(VI)-polluted soil microcosms with the bacterial consortium was investigated using the best factor levels. Contaminated soil by 20 and 60 mg/Kg of Cr(VI) showed reductions of 83% and 65% of initial Cr(VI) by the bacterial consortium, suggesting that this bacterial consortium might diminish phytoavailable Cr(VI) in soil and be useful for cleaning up chromium-contaminated sites.     

Advanced kinetic model of the Cr(VI) removal by biomaterials at various pHs and temperatures

Recently, a new and simple kinetic model was derived from a basic concept of the redox reaction between Cr(VI) and biomaterials, and successfully described the removal behavior of Cr(VI) under various Cr(VI) and biomaterial concentrations. However, this model did not consider the effects of pH and temperature on the Cr(VI) removal by biomaterials. In this study, a new efficient biomaterial, pine needle, capable of removing Cr(VI) was used as a model one to study the Cr(VI) removal by biomaterials. Analysis of chromium species in aqueous and solid phases revealed that the removal mechanism of Cr(VI) by pine needle was its reduction into Cr(III). The removal rate of Cr(VI) increased with a decrease in pH or with an increase of temperature. Finally, an advanced kinetic model in the form of -d[Cr(VI)]/dt = Ae(Ea/RT)[H+]n[Cr(VI)][OCs] was derived, and successfully predicted the time-dependent Cr(VI) concentration at various pHs (2-4) and temperatures (10-55 degrees C).     

Short-term batch studies on biological removal of chromium from synthetic wastewater using activated sludge biomass

Biological treatment of Cr(VI)-containing wastewater has drawn much attention recently as a method to treat environmental Cr(VI) contamination. The activated sludge method, due to its convenient operation and easy-to-scale-up, has been widely applied to treat municipal wastewater and some industrial wastewaters. In order to develop a suitable technique using activated sludge as the biomass to continuously remove Cr(VI) from wastewater, this paper investigated in short-term batch experiments the environmental elements affecting chromium biological removal from synthetic wastewater. The dissolved oxygen (DO), Cr(VI) initial concentration, biomass density, temperature, glucose content in the influent and contact time were observed to strongly influence chromium removal. It was found that the chromium removal efficiency decreased with the increase of DO and Cr(VI) initial concentration as well as glucose content in the feed, but increases in temperature and contact time improved the chromium removal efficiency. Although raising biomass concentration resulted in an increased chromium removal efficiency under both anaerobic and aerobic conditions, its influence on specific chromium removal was not significant.     

Removal of hexavalent chromium in tannery wastewater by Bacillus cereus

Bacillus cereus was used to remove chromium (Cr(VI)) from medium containing tannery wastewater under different conditions. The maximum rate of Cr(VI) removal was attained at a temperature of 37?°C, pH of 7.0-9.0, and biomass of 20 g/L when the initial Cr(VI) concentration was less than 50?mg/L. Under the optimum conditions, the Cr(VI) in tannery wastewater was treated with each cellular component of B. cereus to detect its ability to reduce Cr(VI). The results showed that the removal rate of Cr(VI) for the cell-free extracts could reach 92.70%, which was close to that of the whole cells (96.85%), indicating that the Cr(VI) reductase generated by B.?cereus is primarily intracellular. Additionally, during continuous culture of the B. cereus, the strain showed good consecutive growth and removal ability. After treatment of 20?mg/L Cr(VI) for 48?h, the B. cereus was observed by SEM and TEM-EDX. SEM images showed that the B.?cereus used to treat Cr(VI) grew well and had a uniform cellular size. TEM-EDX analysis revealed large quantities of chromium in the B. cereus cells used to treat Cr(VI). Overall, the results presented herein demonstrate that B. cereus can be used as a new biomaterial to remove Cr(VI) from tannery wastewater.     

Removal of chromium from industrial waste by using eucalyptus bark

Several low cost biomaterials such as baggase, charred rice husk, activated charcoal and eucalyptus bark (EB) were tested for removal of chromium. All the experiments were carried out in batch process with laboratory prepared samples and wastewater obtained from metal finishing section of auto ancillary unit. The adsorbent, which had highest chromium(VI) removal was EB. Influences of chromium concentration, pH, contact time on removal of chromium from effluent was investigated. The adsorption data were fitted well by Freundlich isotherm. The kinetic data were analyzed by using a first order Lagergren kinetic. The Gibbs free energy was obtained for each system and was found to be -1.879 kJ mol(-1) for Cr(VI) and -3.885 kJ mol(-1) for Cr(III) for removal from industrial effluent. The negative value of deltaG0 indicates the feasibility and spontaneous nature of adsorption. The maximum removal of Cr(VI) was observed at pH 2. Adsorption capacity was found to be 45 mg/g of adsorbent, at Cr(VI) concentration in the effluent being 250 mg/l. A waste water sample containing Cr(VI), Cr(III), Mg, and Ca obtained from industrial unit showed satisfactory removal of chromium. The results indicate that eucalyptus bark can be used for the removal of chromium.     

Uptake of chromium(III) and chromium(VI) compounds in the yeast cell structure

The study presented in this article investigated the influence of different Cr(III) and Cr(VI) compounds in the cultivation medium on the uptake and localization of chromium in the cell structure of the yeast Candida intermedia. The morphology of the yeast cell surface was observed by the scanning electron microscopy. Results demonstrated that the growth inhibitory concentration of Cr(III) in the cultivation medium induced changes in the yeast cell shape and affected the budding pattern, while inhibitory concentration of Cr(VI) did not cause any visible effects on morphological properties of the yeast cells. The amount of total accumulated chromium in yeast cells and the distribution of chromium between the yeast cell walls and spheroplasts were determined by atomic absorption spectroscopy. No significant differences were found neither in total chromium accumulation nor in the distribution of chromium in yeast cell walls and spheroplasts between the two of Cr(VI) compounds. Conversely, substantial differences between Cr(III) compounds were demonstrated in the total uptake as well as the localization of chromium in yeast cells.     

Aerobic Reduction of Hexavalent Chromium in Soil by Indigenous Microorganisms

Surface soil containing 25,100 mg/kg total Cr [12,400 mg/kg Cr(VI)] obtained from a Superfund site was used in laboratory microcosm studies to evaluate the potential for aerobic reduction of Cr(VI) by the indigenous soil microbial community. Hexavalent chromium in soil was reduced by as much as 33% (from 1840 to 1240 mg/L) within 21 days under enrichment conditions. Reduction of Cr(VI) in this system was biologically mediated and depended on the availability of usable energy sources. Mass balance studies suggested that the microbial populations removed Cr(VI) from the soil solutions by reduction. Indigenous microbial soil communities even with no history of Cr(VI) contamination were capable of mediating this process. However, Cr(VI) removal was not observed when microbial populations from a sewage sludge sample were added to the soil microcosms. The results suggest that Cr(VI)-reducing microbial populations are widespread in soil, and thus the potential exists for in situ remediation of environmentally significant levels of Cr( VI) contamination.     

Hexavalent chromium removal in vitro and from industrial wastes, using chromate-resistant strains of filamentous fungi indigenous to contaminated wastes

Two chromate-resistant filamentous fungi, strains H13 and Ed8, were selected from seven independent fungal isolates indigenous to Cr(VI)-contaminated soil because of their ability to decrease hexavalent chromium levels in the growth medium. Morphophysiological studies identified strain H13 as a Penicillium sp. isolate and Ed8 as an Aspergillus sp. isolate. When incubated in minimal medium with glucose as a carbon source and in the presence of 50 microg/mL Cr(VI), these strains caused complete disappearance of Cr(VI) in the growth medium after about 72 h of incubation. Total chromium concentration in growth medium was constant during culture growth, and no accumulation of chromium in fungal biomass was observed. Quantitative determinations of oxidized and reduced chromium species during the reduction process revealed stoichiometric conversion of Cr(VI) to Cr(III). A decrease in Cr(VI) levels from industrial wastes was also induced by Ed8 or H13 biomass. These results indicate that chromate-resistant filamentous fungi with Cr(VI)-reducing capability could be useful for the removal of Cr(VI) contamination.     

Microbial reduction of Cr(VI) in the presence of chromate conversion coating constituents

The reduction of Cr(VI) by the metal-reducing bacterium Shewanella oneidensis MR-1 was evaluated, to determine the potential for exploiting Cr(VI) bioreduction as a means of treating chromate conversion coating (CCC) waste streams. Inclusion of Cr(VI) at concentrations ≥1 mM inhibited aerobic growth of S. oneidensis, but that organism was able to reduce Cr(VI) at a concentration of up to 1 mM under anaerobic, nongrowth conditions. S. oneidensis reduced Cr(VI) in the presence of common CCC constituents, with the exception of ferricyanide, when these CCC constituents were included at concentrations typical of CCC waste streams. Ferricyanide inhibited neither aerobic growth nor metabolism under aerobic, nitrate- or iron-reducing conditions, suggesting that the ferricyanide-depended inhibition of Cr(VI) reduction is not due to broad metabolic inhibition, but is specific to Cr(VI) reduction. Results indicate that under some conditions, the activities of metal-reducing bacteria, such as S. oneidensis, could be exploited for the removal of Cr(VI) from CCC waste streams under appropriate conditions.     

Chromium(III) Oxidation Coupled with Microbially Mediated Mn(II) Oxidation

Abstract

Reductive immobilization of Cr(VI) has been widely explored as a cost-effective approach for Cr-contaminated site remediation. In soils containing manganese oxides, however, the immobilized form of chromium, i.e., Cr(III), could potentially be reoxidized. In this study, batch experiments were conducted to assess whether there were any microbial processes that could accelerate Cr(III) oxidation in aerobic, manganese-containing systems. The results showed that in the presence of at least one species of manganese oxidizers, Pseudomonas putida, Cr(III) oxidation took place at low concentrations of Cr(III). About 30–50% of added Cr(III) (10–200 μ M) was oxidized to Cr(VI) within five days in the systems with P. putida and biogenic Mn oxides. The rate of Cr(III) oxidation was approximately proportional to the initial concentration of Cr(III) up to 100 μ M, but the growth of P. putida was partially inhibited by Cr(III) at 200 μ M and totally stopped when it reached 500 μ M. Cr(III) oxidation was dependent upon the biogenic formation of Mn oxides, though the oxidation rate was not directly proportional to the amount of Mn oxides formed. Chromium(III) oxidation took place through a catalytic pathway, in which the microbes mediated Mn(II) oxidation to form Mn-oxides, and Cr(III) was subsequently oxidized by the biogenic Mn-oxides.     

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

A comparative study has been made on the sensitivity of the yeast Pichia guilliermondii to Cr (III) and Cr (VI) as well as on the Cr uptake potential at growth-inhibitory concentrations of chromium. The strains used in the study were either isolated from natural sources or obtained from a laboratory strain collection. The results show that most of the natural strains were more tolerant to chromium and were able to grow in the presence of 5 mM Cr (III) or 0.5 mM Cr (VI), that is at concentrations which substantially inhibited the growth of laboratory strains. The cellular Cr content after treatment was similar for both strain types and ranged from 1.2-4.0 mg/g d.w. and 0.4-0.9 mg/g d.w., for Cr (III) and Cr (VI) forms, respectively, however, in one case of a natural strain it reached the value of 10 mg Cr (III)/g dry mass. Natural-source strains were grouped into four groups based on the yeasts' differential response to Cr (III) and Cr (VI). Hexavalent Cr-resistant mutants of a P. giuilliermondii laboratory strain, which revealed markedly changed capabilities of chromium accumulation, were obtained by means of UV-induced mutagenesis. Cr (VI) treatment triggered oversynthesis of riboflavin and the addition of exogenous riboflavin increased P. guilliermondii resistance to both Cr (III) and Cr (VI). Electrophoretic protein profiles revealed the induction and/or suppression of several proteins in response to toxic Cr (VI) levels.