Heavy metal accumulation by Saccharomyces cerevisiae cells armed with metal binding hexapeptides targeted to the inner face of the plasma membrane
- 840 Downloads
Accumulation of heavy metals without developing toxicity symptoms is a phenotype restricted to a small group of plants called hyperaccumulators, whose metal-related characteristics suggested the high potential in biotechnologies such as bioremediation and bioextraction. In an attempt to extrapolate the heavy metal hyperaccumulating phenotype to yeast, we obtained Saccharomyces cerevisiae cells armed with non-natural metal-binding hexapeptides targeted to the inner face of the plasma membrane, expected to sequester the metal ions once they penetrated the cell. We describe the construction of S. cerevisiae strains overexpressing metal-binding hexapeptides (MeBHxP) fused to the carboxy-terminus of a myristoylated green fluorescent protein (myrGFP). Three non-toxic myrGFP-MeBHxP (myrGFP-H6, myrGFP-C6, and myrGFP-(DE)3) were investigated against an array of heavy metals in terms of their effect on S. cerevisiae growth, heavy metal (hyper) accumulation, and capacity to remove heavy metal from contaminated environments.
KeywordsHeavy metal Metal-binding hexapeptide Accumulation Saccharomyces cerevisiae
The research leading to these results has received funding from the Romanian—EEA Research Program operated by the Ministry of National Education under the EEA Financial Mechanism 2009-2014 and Project Contract No 21 SEE/30.06.2014.
Compliance with ethical standards
This study was funded by the EEA Financial Mechanism 2009–2014 (Contract No 21 SEE/30.06.2014).
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Amberg DC, Burke DJ, Strathern JN (2005) “Quick and dirty” plasmid transformation of yeast colonies. In: Burke D, Dawson D, Stearns T (eds) Methods in yeast genetics. A Cold Spring Harbor laboratory course manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, pp 113–114Google Scholar
- Bradl H (ed) (2002) Heavy metals in the environment: origin, interaction and remediation, vol 6. Academic, LondonGoogle Scholar
- Dürr G, Strayle J, Plemper R, Elbs S, Klee SK, Catty P, Wolf DH, Rudolph HK (1998) The medial-Golgi ion pump Pmr1 supplies the yeast secretory pathway with Ca2+ and Mn2+ required for glycosylation, sorting, and endoplasmic reticulum-associated protein degradation. Mol Biol Cell 9:1149–1162CrossRefPubMedPubMedCentralGoogle Scholar
- Feldmann H (ed) (2012) Transition metal transport. In Yeast: molecular and cell biology, 2nd edn. Wiley-Blackwell, Hoboken, pp 226–232Google Scholar
- Guthrie C, Fink GR (eds) (1991) Guide to yeast genetics and molecular biology. Methods Enzymol 194:1–863Google Scholar
- Kuroda K, Ebisutani K, Iida K, Nishitani T, Ueda M (2014) Enhanced adsorption and recovery of uranyl ions by NikR mutant-displaying yeast. Biomol Ther 4:390–401Google Scholar
- Shibasaki S, Ueda M, Iizuka T, Hirayama M, Ikeda Y, Kamasawa N, Osumi M, Tanaka A (2001) Quantitative evaluation of the enhanced green fluorescent protein displayed on the cell surface of Saccharomyces cerevisiae by fluorometric and confocal laser scanning microscopic analyses. Appl Microbiol Biotechnol 55:471–475CrossRefPubMedGoogle Scholar
- Shibasaki S, Kuroda K, Duc Nguyen H, Mori T, Zou W, Ueda M (2006) Detection of protein-protein interactions by a combination of a novel cytoplasmic membrane targeting system of recombinant proteins and fluorescence resonance energy transfer. Appl Microbiol Biotechnol 70:451–457CrossRefPubMedGoogle Scholar