Descrizione del progetto
Sequenza proteica e funzione di enzimi non caratterizzati nei funghi
I rapidi progressi della tecnologia di sequenziamento del genoma lasciano circa il 30 % delle proteine previste come «orfane», nel senso che non ci sono informazioni sulla loro funzione o sugli enzimi strettamente correlati. I funghi sono noti come produttori prolifici di metaboliti secondari, tuttavia lo studio dei cluster genici biosintetici all’origine degli stessi è ancora scarsamente sviluppato. Il progetto fORPHAN, finanziato dall’UE, mira a sviluppare un percorso di deorfanizzazione basato sulla caratterizzazione computazionale e sperimentale degli enzimi del regno fungino. I ricercatori propongono di usare ricerche approfondite di database mirati per identificare gli enzimi biosintetici chiave nei genomi dei funghi disponibili al pubblico. I promettenti candidati genici saranno espressi in vitro e in ospiti microbici ricombinanti per caratterizzare la loro attività su una gamma di substrati e mediante cristallografia a raggi X.
Obiettivo
Fast advances in genome sequencing and sequence processing technology leave ~30% of predicted proteins as orphans, meaning without known function or closely related enzymes. Being able to assign a function to such orphans opens avenues to select for and design powerful biocatalysts – individual enzymes, biosynthetic pathways or entire organisms. The herein proposed research aims at developing a de-orphanizing pipeline “fORPHAN” based on computational and experimental characterization of enzymes from the fungal kingdom. Fungi are known to be prolific producers of secondary metabolites, however, the study of the underlying biosynthetic gene clusters is still a fairly young field. The use of automated search and annotation pipelines is currently limited by the small number of experimentally characterized genes and gene clusters that can be used to train the algorithms. Therefore, I propose to use deep targeted database searches to identify key biosynthetic enzymes in the publicly available genomes of fungi. First targets will be the thus-far uncharacterized family of putative chalcone isomerases and the recently discovered family of type III polyketide synthases, which both have great potential for in vivo and in vitro applications. Several gene candidates will be expressed in vitro and in recombinant microbial hosts, and tested for activity on a range of substrates. Interesting candidates will also be characterized by X-ray crystallography. This research will not only yield biotechnologically-relevant catalysts but also provide the bioinformatic foundation for more challenging genome mining projects.
Campo scientifico
- natural sciencesearth and related environmental sciencesgeologymineralogycrystallography
- natural sciencesbiological sciencesmicrobiologymycology
- natural scienceschemical sciencescatalysisbiocatalysis
- natural sciencesbiological sciencesgeneticsgenomes
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes
Parole chiave
Programma(i)
Argomento(i)
Meccanismo di finanziamento
MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)Coordinatore
9712CP Groningen
Paesi Bassi