Microbal Degradation of Tauropine. An investigation

Praktikumsbericht / -arbeit , 2015 , 15 Seiten , Note: 1.0

Chemie - Biochemie

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.

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

• Hochschule: Universität Konstanz
• Note: 1.0
• Autor: Manuel Langer (Autor:in)
• Erscheinungsjahr: 2015
• Seiten: 15
• Katalognummer: V335477
• ISBN (eBook): 9783668255654
• ISBN (Buch): 9783668255661
• Sprache: Englisch
• Schlagworte: Tauropine, Microbal Degradation, microoorganism, dehydrogenase, metabolization