Extreme environments created through natural and anthropogenic processes harbor microbes with diverse physiologies capable of catalyzing chemical reactions which are environmentally beneficial on local and global scales. This work focused on two unique environments, the Gulf of Mexico (GoM) submarine mud volcano systems and the subsurface soils at the Department of Energy s (DOE) Field Research Center (FRC) located in the Oak Ridge National Laboratory Reservation (Oak Ridge, TN). In addition to the physical and chemical extremes present within mud volcano sediments and FRC subsurface soils, these environments are sources of greenhouse gases as well as metal/radionuclide contaminants, respectively. Within the previously uncharacterized mud volcano cold seep sediments, culture-independent analyses of microbial community structure via DNA and RNA clone libraries indicated Gammaproteobacteria and anaerobic methane oxidizing Archaea as the dominant methane oxidizing taxa. Culture-dependent studies of FRC subsurface Arthrobacter and Bacillus isolates demonstrated extensive lateral gene transfer of the PIB-type ATPase metal resistance genes. Additionally, FRC Bacillus and Rahnella isolates demonstrated U(VI) sequestration capabilities as up to 95% soluble U(VI) was immobilization via biogenic phosphate mineral production resulting from constitutive nonspecific phosphohydrolase activity. Findings from these studies identify the prokaryotic diversity within aquatic and terrestrial sediments that contribute to the geochemical cycling of carbon, metals, and radionuclides.

Includes bibliography / xiii, 126 leaves : ill ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, 1989

Thesis (M.S.)--Oregon State University, 2007. / Printout. Includes bibliographical references (leaves 81-83). Also available on the World Wide Web.

Thesis (M.S.)--Duquesne University, 2006. / Title from document title page. Abstract included in electronic submission form. Includes bibliographical references (p. v-xi) and index.

Master of Science

This thesis is focussed on the biosynthesis of three toxins: fluoroacetate, 4-fluoro-L-theronine and β-ethynyl-L-serine which are biosynthesised by the soil bacteria Streptomyces cattleya. The two fluorinated metabolites originate from a common biosynthetic pathway and the thesis describes studies carried out on an aldose-ketose isomerase enzyme of the pathway. The biosynthetic origin of β-ethynyl-L-serine is not known. A total synthesis of this acetylenic amino acid is descibed along with the development of a new analytical method for identifying the metabolite and for future isotope-labelling based biosynthetic studies. Chapter 1 presents the background of this research. It is focussed on the biosynthesis of fluoroacetate and 4-fluoro-L-threonine by S. cattleya and it also introduces alkyne-containing natural products and their biosynthesis. Chapter 2 describes the work carried out on crystallisation of the aldose-ketose isomerase of the fluorometabolite pathway in S. cattleya. Crystals of the isomerase were obtained and they were diffracted by X-ray, however a structure could not be solved. Chapter 3 contains site-directed mutagenesis studies of the isomerase from S. cattleya. Chapter 4 describes an enantioselective total synthesis of β-ethynyl-L-serine. A robust analytical technique based on derivatisation using 'Click' chemistry and LC-MS was developed for the detection of this amino acid directly from the fermentation broth. Chapter 5 details the experimental procedures for compounds synthesised in this thesis and the biological procedures for gene cloning and protein purification.