Isolation of cobalt hyper-resistant mutants of Saccharomyces cerevisiae by in vivo evolutionary engineering approach


Çakar Z. P., ALKIM C., TURANLI B., TOKMAN N., AKMAN S., SARIKAYA M., ...Daha Fazla

JOURNAL OF BIOTECHNOLOGY, cilt.143, sa.2, ss.130-138, 2009 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 143 Sayı: 2
  • Basım Tarihi: 2009
  • Doi Numarası: 10.1016/j.jbiotec.2009.06.024
  • Dergi Adı: JOURNAL OF BIOTECHNOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.130-138
  • Anahtar Kelimeler: Cobalt resistance, Evolutionary engineering, Population heterogeneity, Saccharomyces cerevisiae, ESCHERICHIA-COLI, STRESS, YEAST, PROTEINS, TOLERANT, GLUCOSE, GENE, IRON, STRAINS, GROWTH
  • İstanbul Teknik Üniversitesi Adresli: Evet

Özet

Cobalt is an important element with magnetic properties used in various industrial applications. but is also needed for biological activity. Very little is known about the cellular response of living systems to cobalt stress. Towards investigating this mechanism, we isolated individual Saccharomyces cerevisiae cells resistant to high cobalt concentrations up to 8 mmol l(-1). by employing four different 'in vivo' evolutionary engineering strategies: selection under constant or gradually increasing stress levels, and selection under continuous or pulse exposure to cobalt stress. Selection under continuous exposure to gradually increasing cobalt stress levels yielded the most resistant cell population to cobalt. However, the resistance was highly heterogeneous within the mutant populations ranging from 3- to 3700-fold survival rate of isolated individuals to 8 mmol l(-1) CoCl(2) in the most resistant population. Moreover, cobalt-resistant individual colonies were associated with 2-4-times lower intracellular cobalt contents as compared to wild-type, and with cross-resistance to metals such as nickel, zinc, manganese, but not to copper and chromium ions. Contrary to mutants evolved under continuous exposure to cobalt, those isolated by pulse exposure strategy also exhibited resistance to heat shock and hydrogen peroxide stress. Taken together, this study reinforced the fact that evolutionary engineering is useful in selecting strains with very specific phenotypes, and further illustrated the importance of the strategy chosen to isolate the best evolved strain. (C) 2009 Elsevier B.V. All rights reserved.