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.
Inhaltsverzeichnis (Table of Contents)
- Introduction
- Opines
- 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 tauropine
- 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)
Zielsetzung und Themenschwerpunkte (Objectives and Key Themes)
This study aims to investigate the microbial metabolism of tauropine, a molecule previously well-understood in marine invertebrates but not in microorganisms. The research focuses on identifying the enzymes involved in tauropine degradation, characterizing the degradation pathway, and exploring the potential for antibacterial and antifungal activity in associated strains.
- Microbial degradation pathway of tauropine
- Identification and characterization of tauropine dehydrogenase in microorganisms
- Downstream metabolic processes following tauropine degradation
- Antibacterial and antifungal activity screening of isolated strains
- Comparison of microbial tauropine metabolism with that in marine invertebrates
Zusammenfassung der Kapitel (Chapter Summaries)
Introduction: This chapter introduces tauropine, a C5-amino sulfonate belonging to the opine group, and its known role in the anaerobic metabolism of marine invertebrates. It highlights the established understanding of tauropine degradation in these organisms, emphasizing the role of tauropine dehydrogenase. The chapter then contrasts this with the lack of knowledge regarding tauropine metabolism in microorganisms, posing the central research question of the study: how is tauropine metabolized by bacteria? The introduction sets the stage by highlighting the significance of this gap in knowledge and outlines the main objectives of the research.
Opines: This section provides background information on opines, a class of molecules formed by reductive condensation. It discusses the historical context of opine discovery, focusing on D-octopine and its role as a functional analog of lactate. The section expands on the role of opines in anaerobic metabolism in lower invertebrates, illustrating their importance in maintaining cellular redox balance. It establishes the broader context of tauropine within this class of molecules, setting the stage for a detailed investigation into its microbial degradation.
Results and Discussion: This section presents the findings of the experimental investigations into the microbial degradation of tauropine. It details the proposed dissimilation pathway, the detection of metabolites, and the identification of enzymes involved. The results likely include data from various experiments, such as SDS-PAGE analysis, enzyme activity assays, and 16S rDNA sequencing. The significance of the findings is discussed in relation to the initial research questions, potentially revealing similarities and differences compared to the well-established invertebrate pathways. The synthesis and isolation of tauropine and potential degrading strains are also discussed, providing crucial experimental detail and contextualizing the main findings.
Methods: This chapter outlines the experimental procedures used in the study. It details the synthesis, purification, and identification of tauropine; the isolation and cultivation of tauropine-degrading bacterial strains; the 16S rDNA analysis for identification; antibacterial and antifungal activity screening methods; growth curve analysis; cell-free extract preparation; and enzymatic activity tests. This chapter provides the necessary methodological transparency to assess the rigor and reproducibility of the study's findings.
Schlüsselwörter (Keywords)
Tauropine, microbial degradation, opines, tauropine dehydrogenase, anaerobic metabolism, marine invertebrates, bacteria, Ralstonia solanacearum, 16S rDNA, SDS-PAGE, enzyme activity, antibacterial activity, antifungal activity.
Frequently Asked Questions: Microbial Metabolism of Tauropine
What is the main topic of this study?
This study investigates the microbial metabolism of tauropine, a molecule previously well-understood in marine invertebrates but not in microorganisms. The research focuses on identifying the enzymes involved in tauropine degradation, characterizing the degradation pathway, and exploring the potential for antibacterial and antifungal activity in associated strains.
What are the key objectives of this research?
The key objectives include: identifying the microbial degradation pathway of tauropine; identifying and characterizing tauropine dehydrogenase in microorganisms; understanding downstream metabolic processes following tauropine degradation; screening isolated strains for antibacterial and antifungal activity; and comparing microbial tauropine metabolism with that in marine invertebrates.
What is tauropine?
Tauropine is a C5-amino sulfonate belonging to the opine group. It's known to play a role in the anaerobic metabolism of marine invertebrates.
What are opines?
Opines are a class of molecules formed by reductive condensation. The study provides background on opines, focusing on their role in anaerobic metabolism in lower invertebrates and their importance in maintaining cellular redox balance.
What are the key findings of the study (in general terms)?
The "Results and Discussion" section details the proposed dissimilation pathway of tauropine, the detection of metabolites, and the identification of enzymes involved in its degradation. The results likely include data from SDS-PAGE analysis, enzyme activity assays, and 16S rDNA sequencing. The findings are discussed in relation to the initial research questions, comparing microbial and invertebrate pathways.
What methods were used in this study?
The methods included the synthesis, purification, and identification of tauropine; isolation and cultivation of tauropine-degrading bacterial strains; 16S rDNA analysis; antibacterial and antifungal activity screening; growth curve analysis; cell-free extract preparation; and enzymatic activity tests using SDS-PAGE.
What specific enzymes were investigated?
The study focuses on identifying and characterizing tauropine dehydrogenase, a key enzyme in the degradation pathway.
Were any specific bacterial strains investigated?
The study mentions Ralstonia solanacearum in relation to screening for antibacterial and antifungal activity.
What is the significance of the 16S rDNA analysis?
The 16S rDNA analysis was used for the identification of the model organisms (tauropine-degrading strains).
What is the significance of SDS-PAGE?
SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) was used to identify the tauropine dehydrogenase.
What are the chapter summaries?
The document provides chapter summaries for the Introduction (introducing tauropine and the research question), Opines (background on opines and their role in metabolism), Results and Discussion (presenting the findings of the experimental investigation), and Methods (detailing the experimental procedures).
What are the keywords associated with this research?
Keywords include: Tauropine, microbial degradation, opines, tauropine dehydrogenase, anaerobic metabolism, marine invertebrates, bacteria, Ralstonia solanacearum, 16S rDNA, SDS-PAGE, enzyme activity, antibacterial activity, antifungal activity.
- Quote paper
- Manuel Langer (Author), 2015, Microbal Degradation of Tauropine. An investigation, Munich, GRIN Verlag, https://www.hausarbeiten.de/document/335477
