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Identifier

000360484

Title

Απομόνωση και χαρακτηρισμός του ενζύμου λυάση του υδροϋπεροξειδίου των λιπαρών οξέων, υπερεκφρασμένο σε βακτήρια Escherichia coli

Alternative Title

Isolation and characterization of the enzyme hydroperoxide lyase, overexpressed in Escherichia coli bacteria

Author

Παναγάκου, Ιωάννα Βασιλείου

Thesis advisor

Γανωτάκης, Δημήτριος

Abstract

Fatty acid hydroperoxide lyase (HPL), is an important factor of the lipoxygenase path, that is the defense system of plants against exogenous factors. HPL’s function leads to small molecular weight products, of great importance, such as 2-(E)-hexenal and hexanal. In this work, HPL was isolated from E. coli bacteria in which it was cloned, using two different types of nonionic detergents, Triton X-100 and Dodecyl Maltoside (DM). The results of this work showed that, DM is the most appropriate detergent, as it allows further analysis of the protein using spectroscopic techniques. Afterwards, HPL was characterized using circular dichroism and mass spectroscopy, while an attempt isolating the protein directly from olive leaves was carried out. The protein, which was isolated and identified, was a similar in action protein, a peroxidase. Finally, 2-(E)-hexenal and hexanal, two of the HPL’s products, were tested for their antimicrobial, antifungal and anticancer properties. 2-(Δ)-hexenal has an intense effect, while hexanal is less effective.Fatty acid hydroperoxide lyase (HPL), is an important factor of the lipoxygenase path, that is the defense system of plants against exogenous factors. HPL’s function leads to small molecular weight products, of great importance, such as 2-(E)-hexenal and hexanal. In this work, HPL was isolated from E. coli bacteria in which it was cloned, using two different types of nonionic detergents, Triton X-100 and Dodecyl Maltoside (DM). The results of this work showed that, DM is the most appropriate detergent, as it allows further analysis of the protein using spectroscopic techniques. Afterwards, HPL was characterized using circular dichroism and mass spectroscopy, while an attempt isolating the protein directly from olive leaves was carried out. The protein, which was isolated and identified, was a similar in action protein, a peroxidase. Finally, 2-(E)-hexenal and hexanal, two of the HPL’s products, were tested for their antimicrobial, antifungal and anticancer properties. 2-(Δ)-hexenal has an intense effect, while hexanal is less effective.

Language

Greek

Subject

2-(E)-hexenal

2-(E)-εξενάλη

Circular dichroism

E.coli

Fatty acid hydroperoxide lyase (HPL)

Hexanal

Mass spectroscopy

Peroxidase

Εξανάλη