Microbal Degradation of Tauropine. An investigation E-Book

Manuel Langer

• Herausgeber: GRIN Verlag
• Kategorie: Wissenschaft und neue Technologien
• Sprache: Englisch
• Veröffentlichungsjahr: 2016

Beschreibung

Internship Report from the year 2015 in the subject Chemistry - Bio-chemistry, grade: 1.0, University of Constance, language: English, abstract: One interesting aspect is the involvement and the relevance of one sole enzyme in the microbial tauropine degradation pathway: the tauropine dehydrogenase. Therefore three main questions were studied. The first was to verify the action of a tauropine dehydrogenase in microorganisms. The second step was to further characterize this enzyme by its molecular weight and its localization within bacterial cells. In addition, the degradation pathway downstream of the potential tauropine dehydrogenase should be clarified. Therefore, in this study, the metabolism of tauropine in four different model organisms was investigated. As model organisms a Ralstonia strain from fresh water was used and in addition three terrestrial bacterial strains were isolated. The metabolism of tauropine in microorganisms is not yet clarified. Tauropine, besides other opines, has also been reported in the context of bacteria. In fact, it was found in plants, which were infected by agrobacteria with a virulent Ti plasmid. The resulting genetic modification leads to tumor formation, and the plant is triggered to produce opines. As plants cannot use opines themselves, the opines serve as nutrition for the agrobacteria and other opine-degrading bacterial strains. But so far, compared to marine animal phyla, the intermediate steps in the degradation of tauropine in microorganisms are widely unknown. Preliminary investigation in marine bacteria like Ruegeria pomeroyi DSS-3 and Roseovarius nubinhibens ISM has shown that they can use tauropine as source of carbon and nitrogen. Sulfate thereby occurs as end product. It is possible, that the tauropine degradation in bacteria is analogous to that in invertebrates. This would mean that a dehydrogenase is involved. If in microorganisms tauropine can be degraded into pyruvate and taurine by a tauropine dehydrogenase, it is also possible that taurine is further metabolized in the processes, which are already quite well understood. Those processes could include the taurine dehydrogenase and desulfonation by sulfoacetaldehyde acetyltransferase.

Leseprobe

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Table of Contents

Introduction

Opines

The degradation pathway of tauropine in marine invertebrates is well known

The metabolism of tauropine in microorganisms is not yet clarified

Results and Discussion

Dissimilation pathway of tauropine

Detection of the metabolites of tauropine degradation

Investigation of enzymes involved in the tauropine dissimilation pathway

Identification of the tauropine dehydrogenase in one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis

Synthesis of tauropine4,15

Isolation and identification of potential tauropine-degrading strains

Screening for antibacterial and antifungal activity of Ralstonia solanacearum

Summary and Outlook

Methods

Synthesis, purification, identification, and quantification of tauropine

Isolation and cultivation of tauropine-degrading strains from soil

16S rDNA analysis of the model organisms

Screening for antibacterial and antifungal activity

Growth curves

Preparation of cell-free extract

Enzymatic activity tests for clarification of the tauropine degradation pathway

Identification of the tauropine dehydrogenase in one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)

Appendix

Nucleotide sequences

NMR spectra of tauropine