Thèse de doctorat : Génétique Moléculaire : Nancy 1 : 2007. / Titre provenant de l'écran-titre. Bibliogr.

Zugl.: Stuttgart, Univ., Diss., 2009

Bacterial secondary metabolite biosynthetic pathways are frequently encoded in gene clusters. Genomic sequence information allows the identification of likely biosynthetic clusters based on sequence homology to known proteins. Biochemical characterization of suspected biosynthetic enzymes affords the discovery of pathways which may never be identified by traditional screening approaches. In the work presented here, I, in some cases in collaboration with others, characterize the three intragenomic GTP cyclohydrolase II (GCH II) homologs from Streptomyces coelicolor A3(2) and show that one catalyzes a related but distinct reaction from the other two. The basis for the altered activity is investigated and speaks to the chemical mechanism of not only the unusual enzyme but also to all GCH II enzymes. Further, I investigate two other enzymes found in the same gene cluster as the unusual GCH II. Using biochemical techniques, I show that the product of the unusual GCH II is used as a substrate by a creatinine amidohydrolase homolog. Using structural biology, I show that the third enzyme, a 6-pyruvoyltetrahydropterin synthase (PTPS), can not catalyze the PTPS reaction but is capable of binding a pterin substrate. Finally, I propose that the cluster from S. coelicolor containing the unusual GCH II encodes enzymes for a novel pathway to produce a pterin.

<p>To ensure the sustainability of bioethanol production, major attention has been directed to develop feedstocks which provide an alternative to food-crop biomass. Lignocellulosic (LC) biomass, which is chiefly composed of industrial plant residues, is a carbon-rich reservoir that is presently attracting much attention. However LC material is highly recalcitrant to bioprocessing and requires a mixture of physical and enzymatic pretreatment in order to liberate fermentable sugars. Thermostable enzymes are extremely desirable for use in thermophilic fermentations due to their inherent stability. Hemicellulose, a core constituent of LC, requires a cascade of hemicellulases to stimulate the depolymerisation of its xylan backbone. &alpha / -L-arabinofuranosidase (AFase) increases the rate of lignocellulose biodegradation by cleaving arabinofuranosyl residues from xylan thereby increasing the accessibility of other hemicellulases. Twenty thermophilic Actinomycete isolates were screened for AFase activity using pnp-arabinofuranoside as the substrate. Three strains (ORS #1, NDS #4 and WBDS #9) displayed significant AFase activity and were identified as Streptomyces species with 16S rRNA gene sequence analysis. Genomic DNA was isolated from these strains and a cosmid library constructed in the shuttle vector pDF666. Subsequent functional and PCR-based screening revealed no positive clones.</p>

The root system is a vital and dynamic part of a plant throughout its lifetime. Its spatial distribution is the consequence of multiple interactions with the surrounding soil medium. In particular, the presence of pathogens in soil may influence the development of the plant, especially the below-ground part, in both its physiology and its structure. Studies of diseased plant roots may take different approaches and investigate disease effects at different levels. In this study, two groups of four potato (Solanum tuberosum L.) plants were grown in middle-sized plastic pots in a greenhouse, and their root systems, together with the soil medium (i.e., sieved and autoclaved homogeneous sand), were submitted to computed tomography (CT) scanning every two weeks until 10 weeks after planting. For the "diseased" group, sand was inoculated with Streptomyces scabies EF-35, the causal agent of potato common scab, at the time of planting. Disease effects on tissue density of roots and below-ground organs, space occupancy and complexity were assessed by analyzing the CT scanning data in the spatio-temporal approach. Fluctuations of tissue density over time were different, on average, between the two groups. They were characterized by an increase of density in Week 2 for the diseased group, reflecting a hardening of tissue, and a delayed decrease relative to the "healthy" group, suggesting a slower transfer of energy from the seed potato to growing roots. Space occupancy, which was studied via volumetric growth rates evaluated from CT scan data, and complexity, which was quantified by the fractal dimension estimated from skeletonized 3-D images constructed from CT scan data, also showed differences in the first part of the experiment. Original analytical procedures based on data transformation and curve fitting in histogram analysis of CT numbers were developed to obtain those results. In conclusion, the new approach presented here, which is based on the advanced processing of CT scanning data collected over time on developing plant root systems and below-ground organs, can be recommended for future phytopathological applications.

No description available.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2007. / Härtill 4 uppsatser.

Thesis (Ph. D.)--University of Washington, 1999. / Vita. Includes bibliographical references (leaves 148-159).

Stuttgart, Univ., Diss., 2002.

Darmstadt, Techn. Univ., Diss., 2002. / Dateien im PDF-Format