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. 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.
The
study 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. Following 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.
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