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Tesi etd-07312023-102007


Tipo di tesi
Tesi di laurea magistrale
Autore
RUSSO, MICHELE
URN
etd-07312023-102007
Titolo
In vitro dual trait evolution of a minimal synthetic pyrenoid
Dipartimento
BIOLOGIA
Corso di studi
BIOTECNOLOGIE MOLECOLARI
Relatori
relatore Prof.ssa Giuntoli, Beatrice
supervisore Dott. Küffner, Andreas
Parole chiave
  • Biomolecular condensate
  • CO2 fixation
  • Directed evolution
  • High-throughput protein purification
  • Pyrenoid
  • RuBisCO
Data inizio appello
19/09/2023
Consultabilità
Non consultabile
Data di rilascio
19/09/2063
Riassunto
To overcome the natural limitations in terms of carboxylation/oxygenation trade-off of RuBisCO, nature evolved carbon concentrating mechanisms (CCMs).
In the alga Chlamydomonas reinhardtii, the CCMs consist of diverse components, such as carbonate pumps through the thylakoid membranes and, most importantly, the pyrenoid. The pyrenoid is a non-membrane-bound organelle assembled by the targeted interaction of RuBisCO with the intrinsically disordered protein EPYC1 (essential pyrenoid component 1) through liquid-liquid phase separation. In this work, we develop a protocol for the in vitro directed evolution of protein condensates in order to engineer a functional phase separating RuBisCO-IDP fusion mimicking the pyrenoid. By providing an example of the engineering of two traits in a single protein, we show the potential of this approach in reducing complex phenotypes to a minimal number of independent polypeptides, to facilitate their implementation in vivo.
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