Μελέτη των επαγόμενων πρωτεϊνών του βακτηριακού στελέχους Pseudomonas sp. phDV1 όταν αναπτύσσεται παρουσία αρωματικών ενώσεων
2007
academicJournal
Zugriff:
Pseudomonas sp. phDV1 is a very important phenol degrading microorganism, isolated from an enriched mixed culture from samples of petroleum contaminated soil in Denmark. Most of the aromatic compound degraders convert pollutants to catechol, which is then cleaved either through meta or ortho pathway by enzymes catechol 2,3-dioxygenase and catechol 1,2-dioxygenase, yielding 2-hydroxymuconic semialdehyde and cis, cis-muconate, respectively. The metabolic pathway, through which phenol degradation is achieved in this strain, has been previously identified, using mass spectrometry, to be the “meta” one. In this study the proteomes of bacteria grown in different concentrations of phenol, as well as in different pollutants (p-cresol, m-cresol, toluene) used as sole energy and carbon source, are compared. We focus mainly on membrane proteomes and secondly on cytoplasmic. There are strains able to catabolize aromatic compounds using both pathways (ortho, meta), in respect to the concentration of the pollutant. Our results, after spectrophotometric observation of catechol degradation, indicate that Pseudomonas sp. phDV1 uses only the meta pathway for all the cases that have already been examined. The membrane proteomes are analyzed through two dimensional gel electrophoresis (double SDS-PAGE), using SDS as detergent, plus urea as a chaotropic factor for the first dimension, in 10% T polyacrylamide gel and only SDS for the second dimension, in 16,5% T polyacrylamide gel. For the cytoplasmic protein fraction, isoelectric focusing (IEF) is performed for the first dimension, using standard IPG strips with non linear pH gradient, within pH 3 to 10. And SDS-PAGE is performed for the second dimension of this two dimensional analysis (IEF/SDS-PAGE). In our comparative analysis of the bacterium’s inducible membrane proteins in respect to nutrient changes, differences in expression of outer membrane proteins have been noticed. In the case of different concentration of phenol, used as sole source of energy and carbon, there are ...
Titel: |
Μελέτη των επαγόμενων πρωτεϊνών του βακτηριακού στελέχους Pseudomonas sp. phDV1 όταν αναπτύσσεται παρουσία αρωματικών ενώσεων
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Autor/in / Beteiligte Person: | Μαρκούτσα, Σταυρούλα ; Τσιώτης, Γεώργιος |
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Veröffentlichung: | 2007 |
Medientyp: | academicJournal |
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