Microbiology Commons^™

Novel Microbial Guilds Implicated In N2o Reduction, Guang He

Doctoral Dissertations

N₂O is a long-recognized greenhouse gas (GHG) with potential in global warming and ozone depletion. Terrestrial ecosystems are a major source of N₂O due to imbalanced N₂O production and consumption. Soil pH is a chief modulating factor controlling net N₂O emissions, and N₂O consumption has been considered negligible under acidic conditions (pH <6). In this dissertation, we obtained solids-free cultures reducing N₂O at pH 4.5. Furthermore, a co-culture (designated culture EV) comprising two interacting bacterial population was acquired via consecutive transfer in mineral salt medium. Integrated phenotypic, metagenomic and metabolomic analysis dictated that the Serratia population excreted certain …

Identification Of The Type Eleven Secretion System (T11ss) And Characterization Of T11ss-Dependent Effector Proteins, Alex S. Grossman

Doctoral Dissertations

Host-associated microbes live in dangerous environments as a result of host immune killing, nutrient provisioning, and physiological conditions. Bacteria have evolved a host of surface and secreted proteins to help interact with this host environment and overcome nutrient limitation. The studies included within this dissertation describe the identification of a novel bacterial secretion system which has evolved to transport these symbiosis mediating proteins. This system, termed the type eleven secretion system (T11SS), is present throughout the Gram negative phylum Proteobacteria, including many human pathogens such as Neisseria meningitidis, Acinetobacter baumanii, Haemophilus haemolyticus, and Proteus vulgaris. Furthermore, …

Metabolism In The Colonocyte: Where Bacteria And Diet Meet, Bohye Park

Doctoral Dissertations

The host-microbiome interaction and the functions of microbial-derived metabolites, including butyrate, are important in colon health. Butyrate is the preferred energy source in colonocytes and has multiple physiological functions in the colon. A metabolic shift in colonocytes toward increased glucose utilization from butyrate oxidation is followed by several critical genetic modifications in cancerous colonocytes. Moreover, it has been suggested that the gut-microbiota composition is influenced by environmental factors, such as diets and the host's physiological status. Therefore, an understanding of the role of colonocyte metabolism toward impacting the host-microbiota commensal relationship would be an important step in understanding the functional …

Determining The Ecological And Physiological Factors Contributing To The Competitive Success Of Prochlorococcus In The Oligotrophic Ocean, Benjamin C. Calfee

Doctoral Dissertations

Prochlorococcus is a genus of extremely successful marine cyanobacteria. This success is realized through its pervasive biogeographical range and presence in almost all open ocean environments where it usually it the dominant phytoplankton. Limited capabilities of culturing and genetic manipulation of this organism have resulted in assumptions about this success overwhelmingly based on field observations. These studies have assumed adaptations for resource uptake and utilization in nutrient limited environments to cause dominance of Prochlorococcus over other photosynthetic microbes. In an attempt to definitively explain this through laboratory culture, we developed a culturing system to assay questions of nutrient limitation effects …

Perturbing Fatty Acid Metabolism In Enterococcus Faecalis Disrupts Responses To Exogenous Fatty Acids And The Antibiotic Daptomycin, Rachel D. Johnston

Doctoral Dissertations

Enterococcus faecalis is an opportunistic pathogen. In the host, it is exposed to fatty acids which impact cellular physiology and induce tolerance to the antibiotic daptomycin. To determine the requirements for induction of daptomycin tolerance, I examined the impacts of blocking de novo fatty acid synthesis or protein synthesis and removing cell wall. I observed that removal of the cell wall induced daptomycin tolerance, indicating that peptidoglycan is necessary for daptomycin to function. As specific exogenous fatty acids induce protection against daptomycin in E. faecalis, I also opted to examine whether incorporation of these free fatty acids was necessary …

Regulation, Mechanism Of Action, And Function Of A Small Toxin Protein In Ehec, Bikash Bogati

Doctoral Dissertations

The zor-orz locus identified in the chromosome of Escherichia coli O157:H7 EDL933 is a type I toxin-antitoxin system that consists of two homologous gene pairs zorO-orzO and zorP-orzP. The zor genes encode for small toxin proteins and the orz genes encode for small RNAs. Previously it was shown that overproduction of ZorO results in cell growth stasis whereas co-expression of orzO rescues cellular growth. Within, I demonstrate that in addition to growth inhibition, ZorO overproduction results in membrane depolarization and ATP depletion but does not impact the gross morphology of E. coli. In vivo translation and subsequent impacts …

Probing Structure, Function And Dynamics In Bacterial Primary And Secondary Transporter-Associated Binding Proteins, Shantanu Shukla

Doctoral Dissertations

Substrate binding proteins (SBPs) are ubiquitous in all life forms and have evolved to perform diverse physiological functions, such as in membrane transport, gene regulation, neurotransmission, and quorum sensing. It is quite astounding to observe such functional diversity among the SBPs even when they are restricted by their fold space. Therefore, the SBPs are an excellent set of proteins that can reveal how proteins evolution novel function in a structurally conserved/constrained fold. This study attempts to understand the phenomenon of affinity and specificity evolution in SBPs by combining a set of biochemical, biophysical, and structural studies on the SBPs involved …

Molecular Classification And Uv Tolerance Of Pigmented Antarctic Extremophiles, Aaron M. Perry

Chancellor’s Honors Program Projects

No abstract provided.

Tetrameric Photosystem I: From Initial Discovery And Characterization In Chroococcidiopsis Sp. Ts-821 To Exploration Of Its Distribution And Understanding Of Its Significance In Cyanobacteria, Meng Li

Doctoral Dissertations

Photosystem I (PSI) forms trimeric complexes in most characterized cyanobacteria. We had reported the tetrameric form of PSI in the unicellular cyanobacterium, Chroococcidiopsis sp. TS-821 (TS-821). Using Cryo-EM, a 3D model of the PSI tetramer structure at 11.5 [Angstrom] resolution was obtained and a 2D map within the membrane plane of at 6.1 [Angstrom]. In contrast to the three-fold symmetry in trimeric PSI crystal structure from T. elongatus, two different inter-monomer interactions involving PsaLs are found in the PSI tetramer. Phylogenetic analysis based on PsaL protein sequences shows that TS-821 is closely related to heterocyst-forming cyanobacteria. Additionally, this tetrameric …

The Effects Of Exogenous Fatty Acids On Enterococcus Faecalis Og1rf, Holly Elizabeth Johnson Saito

Doctoral Dissertations

Enterococcus faecalis is a commensal of the mammalian intestine and an opportunistic pathogen that infects various locations in the human host. Specifically, the organism can infect surgical wounds, the urinary tract, and is an agent of endocarditis. Regardless of niche, this organism has access to fatty acid sources in the host, which may influence its ability to survive and cause infection; yet, the effects of exogenous fatty acids were unknown in E. faecalis. We have shown that complex sources of fatty acids such as bile or serum can significantly alter the membrane content and protect E. faecalis from acute …

Biophysical Studies Of Cell Division Protein Localization Mechanisms In Escherichia Coli, Matthew Wayne Bailey

Doctoral Dissertations

How nanometer-scale proteins position accurately within micron-scale bacteria has intrigued both biologists and physicists alike. A critical process requiring precise protein localization is cell division. In most bacteria, cell division starts with the self-assembly of the FtsZ proteins into filaments that form a ring-like structure encircling the cell at its middle, the Z-ring. The Z-ring is a scaffold for additional proteins that synthesize the lateral cell wall which separates the two daughter cells. If division planes are misplaced relative to bacterial chromosomes, also called nucleoids, daughter cells with incomplete genetic material can be produced. In Escherichia coli, research carried out …

Environmentally Driven Orchestration Of Metabolisms By Prochlorococcus Spp., Martin James Szul

Doctoral Dissertations

In the oligotrophic waters of the world’s open oceans physical factors such as pH, salinity, and temperature are generally stable. The nutrient limited conditions as well as the low environmental variability endemic to these ecosystems select for specialists that gain fitness advantages through minimalism, efficiency, and thrift. These physical characteristics are thought to reduce nutrient demand while allowing for constant metabolic activity and growth, but the mechanisms that promote these fitness advantages are currently unknown. To better understand how these physiologies improve selective fitness for the dominant phytoplankton, we observed metabolic parameters under environmental conditions typical to these waters. In …

Physiological Characterization Of Prochlorococcus Under Abiotic Stressors Temperature And Hydrogen Peroxide, Lanying Ma

Doctoral Dissertations

Cyanobacteria of the genus Prochlorococcus are the smallest and most abundant phytoplankters in the ocean. Temperature is a major influence on Prochlorococcus abundance and distribution in the ocean, but the physiological basis for this relationship is not well understood. In other microbes, lipid and fatty acid composition have been shown to be influenced by temperature, and temperature has also been proposed as a relevant factor for setting the elemental allocation in marine phytoplankton. In this study, we found that percentage of fatty acids unsaturation was negatively related with temperature in some Prochlorococcus strains, but this was not universal. Temperature had …

The Role Of Phosphatidylserine And Phosphatidylethanolamine In Candida Albicans Virulence, Sarah Elizabeth Davis

Doctoral Dissertations

In hospitalized patients with neutropenia, Candida albicans is the fourth leading cause of systemic bloodstream infections, which have a mortality rate of approximately 30 %. The phosphatidylserine synthase enzyme of C. albicans, Cho1p, appears to be a good drug target as a mutant lacking this enzyme (the cho1Δ/Δ [null mutant]) is avirulent in animal models of Candida infections and this enzyme is not conserved in humans. We discovered that the loss of phosphatidylserine (PS) synthesis affects C. albicans' expression of the Als3p adhesin, a virulence protein, and loss of PS synthesis also compromises the cell wall, causing increased …

Elucidating The Impact Of Roseophage On Roseobacter Metabolism And Marine Nutrient Cycles, Nana Yaw Darko Ankrah

Doctoral Dissertations

As the most abundant biological entities in marine environments, viruses are an important component of marine food webs. The activity of viruses contributes significantly to the mortality of marine microorganisms, ultimately influencing biological function and chemical composition of aquatic systems by impacting species composition and flow of carbon, nitrogen and other nutrients. Despite the growing recognition that viral activity contributes to marine biogeochemical cycles, the extent to which virus infection reshapes host metabolism and the effect of this alteration on the composition of host lysate remains poorly understood. Additionally, the degree to which natural bacterioplankton communities metabolise the released lysate …

Comparative Genomics Of Microbial Chemoreceptor Sequence, Structure, And Function, Aaron Daniel Fleetwood

Doctoral Dissertations

Microbial chemotaxis receptors (chemoreceptors) are complex proteins that sense the external environment and signal for flagella-mediated motility, serving as the GPS of the cell. In order to sense a myriad of physicochemical signals and adapt to diverse environmental niches, sensory regions of chemoreceptors are frenetically duplicated, mutated, or lost. Conversely, the chemoreceptor signaling region is a highly conserved protein domain. Extreme conservation of this domain is necessary because it determines very specific helical secondary, tertiary, and quaternary structures of the protein while simultaneously choreographing a network of interactions with the adaptor protein CheW and the histidine kinase CheA. This dichotomous …

Novobiocin As An Allosteric Modulator Of Ste2p, Jeffrey K. Rymer, Melinda Hauser, Jeffrey M. Becker

EURēCA: Exhibition of Undergraduate Research and Creative Achievement

G protein-coupled receptors (GPCRs) are the target of 30-50% of all prescribed drugs for human medicine and are therefore the subject of intense study by the scientific community. It has been recognized recently that compounds called allosteric modulators can regulate GPCR activity by binding a GPCR at sites not occupied by the normal receptor-activating molecule. Such allosteric compounds are desirable drug candidates as they may produce fewer toxic side-effects than standard drugs that target GPCRs. The purpose of this study was to determine the interaction of different allosteric modulators with Ste2p, a model GPCR expressed in the yeast Saccharomyces cerevisiae. …

Development And Application Of Liquid Chromatography-Tandem Mass Spectrometry Methods To The Understanding Of Metabolism And Cell-Cell Signaling In Several Biological Systems, Jessica Renee Gooding

Doctoral Dissertations

Liquid chromatography tandem mass spectrometry has become a powerful tool for investigating biological systems. Herein we describe the development of both isotope dilution mass spectrometry methods and targeted metabolomics methods for the study of metabolic and cell-cell signaling applications.

A putative yeast enzyme was characterized by discovery metabolite profiling, kinetic flux profiling, transcriptomics and structural biology. These experiments demonstrated that the enzyme shb17 was a sedoheptulose bisphosphatase that provides a thermodynamically dedicated step towards riboneogenesis, leading to the redefinition of the canonical pentose phosphate pathway.

An extension of metabolic profiling and kinetic flux profiling methods was developed for a set …

Genomic And Molecular Analysis Of The Exopolysaccharide Production In The Bacterium Thauera Aminoaromatica Mz1t, Ke Jiang

Doctoral Dissertations

Thauera aminoaromatica MZ1T is an exopolysaccharide (EPS)-producing Gram negative bacterium isolated from the wastewater treatment plant of a major industrial chemical manufacturer as the causal agent for poor sludge dewatering. It shares common features with other known Thauera spp. (i.e. Thauera aromatica, and Thauera selenatis), being capable of degrading aromatic compounds anaerobically and using acetate and succinate as carbon sources. It is unique among the Thauera spp. in its production of abundant EPS which results in viscous bulking and poor sludge dewaterability. In this respect, it is similar to Azoarcus sp. EbN1 and BH72. Thaueran is the proposed …

The ‘Helper’ Phenotype: A Symbiotic Interaction Between Prochlorococcus And Hydrogen Peroxide Scavenging Microorganisms, James Jeffrey Morris

Doctoral Dissertations

The unicellular cyanobacterium Prochlorococcus is the numerically dominant photosynthetic organism throughout the temperate and tropical open oceans, but it is difficult to grow in pure cultures. We developed a system for rendering spontaneous streptomycin-resistant mutants of Prochlorococcus axenic by diluting them to extinction in the presence of “helper” heterotrophic bacteria, allowing them to grow to high cell concentrations, and then killing the helpers with streptomycin. Using axenic strains obtained in this fashion, we demonstrated that Prochlorococcus experiences a number of growth defects in dilute axenic culture, including reduced growth rate, inability to form colonies on solid media, and higher incidence …

Use Of Proteomics Tools To Investigate Protein Expression In Azospirillum Brasilense, Gurusahai K. Khalsa-Moyers

Doctoral Dissertations

Mass spectrometry based proteomics has emerged as a powerful methodology for investigating protein expression. “Bottom up” techniques in which proteins are first digested, and resulting peptides separated via multi-dimensional chromatography then analyzed via mass spectrometry provide a wide depth of coverage of expressed proteomes. This technique has been successfully and extensively used to survey protein expression (expression proteomics) and also to investigate proteins and their associated interacting partners in order to ascertain function of unknown proteins (functional proteomics). Azospirillum brasilense is a free-living diazotrophic soil bacteria, with world-wide significance as a plant-growth promoting bacteria. Living within the rhizosphere of cereal …