Theses and Dissertations at Montana State University (MSU)
Permanent URI for this collectionhttps://scholarworks.montana.edu/handle/1/733
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Item Identification of cellulolytic hot spring organisms through bioorthogonal labeling(Montana State University - Bozeman, College of Letters & Science, 2021) Reichart, Nicholas John; Chairperson, Graduate Committee: Roland Hatzenpichler; This is a manuscript style paper that includes co-authored chapters.Microbial physiology is the study of the metabolism and function of microorganisms. The recent expansion of genomic diversity has outpaced the description of physiology. To better understand microbial metabolisms and environmental processes, more detailed research is needed for both novel and undescribed microbes. While many new methods are being developed to describe in situ microbial activity, this dissertation implements bioorthogonal non-canonical amino acid tagging as a proxy to track metabolic activity of microbes under close to environment conditions. Using differential analyses on hot spring microbial communities, we were able to show that certain microbial taxa had preferential activity towards specific incubation amendments. Previous activity-based studies had shown that hot springs were a unique environment for discovering cellulolytic microbes that could be used in industrial processing of plant biomass. Herein, we used computational analysis to screen publicly available metagenomic datasets to identify the enzymatic potential of hot springs worldwide. The wide diversity of taxa and biomass degrading enzymes were investigated and hot springs were further highlighted as a system that could be used to find improvement for the industry of plant biomass degradation and processing. To build upon the cellulolytic potential found in hot spring metagenomic datasets, bioorthogonal non-canonical amino acid tagging coupled with fluorescence-activated cell sorting was applied to the biotechnological relevant field of plant biomass degradation to identify microbes involved in the cellulolytic process. Examination of the active microbes revealed difference in the community when supplemented with cellulose. Taken together, the work in this dissertation served to expand and apply the recent development of activity-based studies used to describe environmental microbial populations, with a focus on plant biomass degradation.Item The application of mass spectrometry-based 'omics technologies to investigate environmental interactions of microbial systems(Montana State University - Bozeman, College of Letters & Science, 2015) Tigges, Michelle Marie; Chairperson, Graduate Committee: Brian Bothner; Michelle Tigges and Heidi Smith were main authors, and Juliana D'Andrill, Al Parker, Brian Bothner and Christine Foreman were co-authors of the article, 'Lability of environmental dissolved organic matter drives dynamic microbial processing' submitted to the journal 'Proceedings of the National Academy of Sciences' which is contained within this thesis.Microorganisms interact with their surroundings and each other. However, these interactions are complex and difficult to understand. This research presents the utilization of simplified environmental microbial systems from the extremes of life to gain insight into the roles of microbes in diverse processes including biogeochemical cycling and viral infection. The complex mixture of organic compounds within aquatic systems, known as dissolved organic matter (DOM), is an integral component of the global carbon cycle. It is a carbon source for microbial activity and impacts biogeochemical and ecological processes. However, little is known about the release and bioconversion of these compounds. This thesis presents a liquid chromatography coupled mass spectrometry (LCMS) based exometabolomics approach to chemically characterize the interaction between DOM and the representative microbial species that transform it. This work illustrates for the first time the ability to measure the relative abundance of molecular constituents of DOM during heterotrophic bacterial processing. Processing was shown to be dynamic over time, even with only single organism interactions. A LCMS based proxy was established to predict the lability of DOM carbon sources, and the labile nature of the source was shown to be a significant factor in DOM processing by single organisms. Further, the temporal interaction of two ecologically relevant beta-Proteobacteria with DOM from the Cotton Glacier, Antarctica highlight the importance of understanding the diversity of single organism DOM interactions to interpret community level bacterial interactions. LCMS-based 'omics techniques can also be utilized to characterize the changes in protein expression associated with viral infection of hyperthermophilic archaea. Viral-host interactions in Sulfolobus archaeal systems are poorly understood, and exhibit a diversity of regulation patterns. LCMS-based shotgun proteomics was utilized to characterize the temporal response of Sulfolobus islandicus to infection by Sulfolobus islandicus rod-shaped virus (SIRV2). The strengths and weaknesses of label-free protein quantitation techniques were assessed, enabling the detection of the regulation of SIRV2 proteins over time and identification of key host responses to infection. Together these studies show the impact of LCMS based 'omics technologies in bringing new insights into environmental microbial interactions.Item Factors affecting utilization of aspartic acid by Leuconostoc Mesenteroides P-60(Montana State University - Bozeman, College of Letters & Science, 1950) Clark, Richard S.Item Investigation of biologically active metabolites from symbiotic microorganisms(Montana State University - Bozeman, College of Letters & Science, 1988) Stierle, Andrea AnneItem Free lipids of hot spring microbial mats of possible evolutionary significance(Montana State University - Bozeman, College of Letters & Science, 1988) Shiea, Jentaie