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Identifier 000426013
Title Μελέτη της αποικοδόμησης μεθυλοφαινολών από το βακτήριο Pseudomonas sp. Strain PhDV1
Alternative Title Study of the degradation of methylphenols from the bacterium Pseudomonas sp. Strain PhDV1
Author Βαλσαμίδης, Ιωάννης Σ.
Thesis advisor Τσιώτης, Γεώργιος
Reviewer Γανωτάκης. Δημήτριος
Κοτζαμπάσης, Κυριάκος
Abstract Pseudomonas sp. PhDV1 is a gram-negative bacterium that has the ability to degradade aromatic compounds such phenol and toluene by using the meta metabolic pathway. In this project, it was attempted to grow this specific bacterium by using as a sole carbon and energy source the three methylphenols (cresols). These compounds are also aromatic and their structure is very similar with the phenol and toluene. In this way it was examined if the bacterium has the ability to degradade a wider range of aromatic compounds and also it was vital to determine whether these aromatic compounds are degradaded by the bacterium with the same pathway or it uses a different approach. In order to achieve the growth of the bacterium in these aromatic compounds, different approaches were followed compared to past works, however the growth of the bacterium was successful in all the three isomers of the cresols (ortho-, meta- and para-). The next step was the identification of proteins that are relative with the adaptation and growth of the bacterium in presence of the aromatic compounds and more specifically the enzymes that are related to the metabolic pathway that is used by the bacterium in order to catabolize the methylphenols. This was determined by two different approaches : 1) with the UV-Vis spectroscopy for the characterization of an important enzyme that is used by the bacterium in order to degrade aromatic compounds and 2) with the proteomic analysis, by comparing those proteins that differentiate when the bacterium grows in presence aromatic pollutants compared to a control sample which in this occasion was the succinic acid. The enzyme that was characterized was the catechol 2,3-dioxygenase which is one of the first enzymes that is used during the meta metabolic pathway and it was proven that this enzyme was expressed by the bacterium in all three of the cresols. This is a clear evidence that the meta metabolic pathway is being used by the bacterium in order to degrade these compounds too. For the proteomic approach, at first three biological samples from five different conditions (succinate, phenol, o-cresol, m-cresol and p-cresol) were isolated and a protocol for the solubilization of the proteins was applied and afterwards the method FASP was used to digest the proteins to peptides and they were identified with nLC-ESI-MS/MS. In total, 2953 proteins were identified for all the five different conditions and with the use of the suitable analytical tool (Perseus) it was determined which of the proteins differentiate during the degradation of the aromatic compounds.
Language Greek
Subject Biodegradation
Proteomic analysis
Πρωτεομική ανάλυση
Issue date 2019-11-29
Collection   School/Department--School of Sciences and Engineering--Department of Chemistry--Post-graduate theses
  Type of Work--Post-graduate theses
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