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Learning from extremophilic fungi: proteomics studies of the oxidative stress defense and protein stability with an eye to proteins’ industrial use.

Projektleitung
Tesei Donatella, BOKU Projektleiter/in
Laufzeit:
15.01.2017-14.01.2020
Programm:
Hertha-Firnberg-Programm
Art der Forschung
Grundlagenforschung
Mitarbeiter/innen
Beteiligte BOKU-Organisationseinheiten
Institut für Biotechnologie
Institut für Umweltbiotechnologie
Gefördert durch
Fonds zur Förderung der wissenschaftlichen Forschung (FWF) , Sensengasse 1, 1090 Wien, Österreich
Abstract
Stress resistance and adaptation are crucial factors determining an organism’s survival to a number of biotic and a-biotic factors. The aim of this project is to elucidate the molecular basis for stress tolerance by proteomic methods and to ultimately detect proteins with key roles in counteracting stress insults. To this purpose, two strains of extremotolerant black fungi – i.e. the wild type and the natural non-melanized mutant of the species Knufia chersonesos – will be used as model organisms displaying a remarkable natural aptitude to tolerate
massive ozone rates as well as protein thermo-stability. The recourse to black fungi is based on the extraordinary ability of these organisms to survive life-threatening environmental conditions, which makes them the most stress resistant Eukaryotes known to date. The two strains will enable to study the dynamics of the oxidative stress defense triggered by the exposure to ozone, in an extremotolerant species. Thus, the data obtained will allow shedding light on the oxidative stress, a condition associated with several physiological and also pathological conditions. The understanding of the basis for tolerance in adapted species will possibly aid accomplishing a second objective of the present project: the finding of novel protein candidates – e.g. radicals scavenging and antioxidant producing enzymes – with potential biotechnological and cosmetic applications. Protein stability and degradation tests will be carried out and special focus will be dedicated to enzymatic cascades reactions involved in degradation of polymers such as PBAT. Time points will be selected for the ozone treatment of the fungal strains cultivated in a multi fermenter under controlled parameters, in order to evaluate the effects of short- and long-term exposure. PBAT-mediated induction of cultures grown in liquid culture will be performed to identify polymer degrading enzymes in the extracellular medium. Shotgun quantitative proteomics approaches for simultaneous detection of changes in protein levels and
identification, will be applied to the analysis of both whole-cell proteome and secretome. While the oxidative stress response will be investigated by means of thiol-based redox proteomics (cysTMTRAQ), label-free shotgun proteomics will serve to the identification
Schlagworte
Mikrobiologie; Mykologie;
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