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Identifier

000403578

Title

Ταυτοποίηση ενζύμων από ψυχρόφιλα βακτήρια με πιθανές εφαρμογές στη βιοτεχνολογία

Alternative Title

Identification of enzymes from psychrophilic bacteria with potential applications in biotechnology

Author

Συμεωνίδου, Ανθή Σ.

Thesis advisor

Καφετζόπουλος, Δημήτριος

Abstract

Cold environments are colonized by psychrophiles, organisms that developed adaptation mechanisms in low temperature through their enzymes, the prerequisite to the environmental adaptation. Further investigations in psychrophilic enzymes could lead to better understanding of adaptation mechanisms and evaluate their potential application in biotechnology due to their high activity in low temperature and heat lability. Possessing an Antarctic psychrophilic/psychrotolerant bacterial strain collection, consisting by 262 strains, a high-throughput screening for DNase and alkaline phosphatase activity was carried out followed by DNA sequencing of the most desirable strains. Furthermore, sequences coding for the target enzymes were identified. A gene from TAB110a strain encoding an alkaline phosphatase, displays ~75% and ~30% identity with ECAP and AnP, respectively. Successful cloning of this gene into the pRSETB plasmid and heterologous expression in an E.coli strain yielded protein with 70-80% purity. Further experiments on the catalytic efficiency and heat resistance, could elucidate its putative psychrophilic character. Using a native substrate, like a plasmid DNA, dephosphorylation efficiency could be measured and correlated with other commercial APs in order to determine its potential use in molecular biology applications.

Language

English

Subject

Alkaline phosphatase

NGS

Psychrophilic enzyme

Αλκαλική φωσφατάση

Αλληλούχιση νέας γενιάς

Ψυχρόφιλο ένζυμο

Issue date

2016-11-18

Collection