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Identifier 000382628
Title Απομόνωση του ενζύμου λύαση του υδροϋπεροξειδίου των λιπαρών οξέων από φύλλα και καρπούς ελιάς
Alternative Title Isolation of the enzyne hydroderoxide lyase from olive leaves and fruits
Author Μυτιληνάκη, Ελισάβετ
Thesis advisor Γανωτάκης, Δημήτριος Φ.
Abstract Hydroperoxide lyase is an enzyme of particular importance, part of the lipoxygenase path. The particular biochemical path starts with the formation of polyunsaturated fatty acids hydroperoxides by lipoxygenase which constitute HPL’s substrate. Products of the reaction that lyase of hydroperoxides catalyzes are hexenal end hexanal, substances that belong to “green leaf volatiles” and also other precursors of important bioregulators in plants. In the present work HPL initially was isolated from E.coli bacterial cultures, organism in which the protein had been previously overexpressed. Furthermore, two different methods were tried in order to isolate HPL from olive fruits and a third method for the same purpose this time using olive leaves as a natural source. In all samples isolated by different methods, in order to pin point HPL´s presence, were conducted kinetics tests in presence of the substrate of the enzyme (13-linoleic acid hydroperoxide), tests for the detection of the reaction’s products with 2,4 hydrophenylhydrazine followed by TLC analysis, and PAGE-SDS electrophoresis. Enzyme identification by mass spectroscopy was not possible and kinetics determination spectrometrically was only partly successful. Lastly, satisfactory results was obtained by the products detection test and as for the protocols used better resulted the one that combined olive fruit grinding, incubation in presence of detergent, consequent centrifugation and purification of the plant extract in an ion-exchange column.
Language Greek
Subject E.coli
Green leaf volatiles
Πτητικά πράσινων φύλλων
Issue date 2014-03-28
Collection   School/Department--School of Sciences and Engineering--Department of Chemistry--Post-graduate theses
  Type of Work--Post-graduate theses
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