Abstract
BackgroundYeast strains endowed with robustness towards copper and/or enriched in intracellular Cu might find application in biotechnology processes, among others in the production of functional foods. Moreover, they can contribute to the study of human diseases related to impairments of copper metabolism. In this study, we investigated the molecular and physiological factors that confer copper tolerance to strains of baker's yeasts.ResultsWe characterized the effects elicited in natural strains of Candida humilis and Saccharomyces cerevisiae by the exposure to copper in the culture broth. We observed that, whereas the growth of Saccharomyces cells was inhibited already at low Cu concentration, C. humilis was naturally robust and tolerated up to 1 g · L-1 CuSO4 in the medium. This resistant strain accumulated over 7 mg of Cu per gram of biomass and escaped severe oxidative stress thanks to high constitutive levels of superoxide dismutase and catalase. Both yeasts were then "evolved" to obtain hyper-resistant cells able to proliferate in high copper medium. While in S. cerevisiae the evolution of robustness towards Cu was paralleled by the increase of antioxidative enzymes, these same activities decreased in evolved hyper-resistant Candida cells. We also characterized in some detail changes in the profile of copper binding proteins, that appeared to be modified by evolution but, again, in a different way in the two yeasts.ConclusionsFollowing evolution, both Candida and Saccharomyces cells were able to proliferate up to 2.5 g · L-1 CuSO4 and to accumulate high amounts of intracellular copper. The comparison of yeasts differing in their robustness, allowed highlighting physiological and molecular determinants of natural and acquired copper tolerance. We observed that different mechanisms contribute to confer metal tolerance: the control of copper uptake, changes in the levels of enzymes involved in oxidative stress response and changes in the copper-binding proteome. However, copper elicits different physiological and molecular reactions in yeasts with different backgrounds.
Highlights
Yeast strains endowed with robustness towards copper and/or enriched in intracellular Cu might find application in biotechnology processes, among others in the production of functional foods
Studies on the accumulation of metals in edible microorganisms are of relevance for the production of functional foods enriched in micronutrients and the industrial production of Saccharomyces cerevisiae biomass highly enriched with organic forms of selenium [7,8]
Tolerance towards copper of one Candida and three Saccharomyces strains was first assessed by a drop test on minimal or rich (YPD) solid medium supplemented with increasing concentrations of copper salt (CuSO4) (Figure 1)
Summary
Yeast strains endowed with robustness towards copper and/or enriched in intracellular Cu might find application in biotechnology processes, among others in the production of functional foods. They can contribute to the study of human diseases related to impairments of copper metabolism. Studies on the accumulation of metals in edible microorganisms are of relevance for the production of functional foods enriched in micronutrients (for example the ones about the inclusion of iron, cobalt, copper and manganese in yeast cells [7]) and the industrial production of Saccharomyces cerevisiae biomass highly enriched with organic forms of selenium [7,8]. Microbial cells were recently evolved to improve their resistance towards multiple stresses [13], cobalt [14], iron- and sulfur-compounds [15], alcohols [16]; and to gain the ability to ferment xylose [17] and lactose [18]
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