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Identifier

000403896

Title

Εξερευνώντας τη χημική ποικιλομορφία των τερπενικών δομών χρησιμοποιώντας συνθετική βιολογία και πρωτεϊνική μηχανική

Alternative Title

Εxploring the chemical diversity of terpene structures using synthetic biology and protein engineering

Author

Σεραφειμίδη, Θεανώ Σ.

Thesis advisor

Καλαντίδης, Σωτήριος

Reviewer

Καμπράνης, Σωτήριος
Κοτζαμπάσης, Κυριάκος

Abstract

Natural products have attracted the interest of man due to their unlimited properties and natural origin. However, all these valuable compounds can hardly be isolated from the plants themselves and human ingenuity found a solution to this problem with the help of heterologous production systems, with dominant target organisms the bacterium Escherichia coli and the yeast Saccharomyces cerevisiae. Terpenes are a large subset of natural compounds, with diverse and complex structure which enables each molecule and their properties to be unique. The following compounds were the focus of this project: sclareol, manool, manoyl oxide and miltiradiene. Sclareol is a diterpene which is predominantly used in the perfume composition replacing ambergris from whales. Manool has shown to have selective cytotoxic activity against tumor cells but so far it is used in perfumery at the same way as sclareol. Manoyl oxide is also used in perfumery, but it is also a precursor for the production of forskolin, which has broad medical use, with the most important so far to be the improvement of heart function. Miltiradiene is an olefin that is also a precursor for the production of ferruginol, a molecule with anti-inflammatory and antioxidant properties. With reference to the multiple uses of these molecules research is carried out with the effort focused in increasing heterologous production of terpenes in yeast using mutagenesis and genetic engineering. In this work transformations of various yeast strains were carried out, that could be used in the future under different growing conditions to optimize production, a mutation of two genes (CDS-Y95 / CLS-Y95) was carried out in order to boost the metabolic path in the direction of interest (diterpene production) and finally there was a sub cloning of the genes CDS-Y95/CLS-Y95/SCLS/p1326 in expression vector pESC-URA from the previously used pYES and pHTDH in order to increase production of the molecules : sclareol, manool, manoyl oxide and miltiradiene.

Language

Greek

Subject

Manool

Manoyl oxide

Miltiradiene

Sclareol

Yeast

Μανόλη

Μιλτιραδιένη

Οξείδιο της Μανόλης

Σκλαρεόλη