Scholarship & Research
Permanent URI for this communityhttps://scholarworks.montana.edu/handle/1/1
Browse
6 results
Search Results
Item Spectral, kinetic, and thermodynamic properties of Cu(I) and Cu(II) binding by methanobactin from Methylosinus trichosporium OB3b(2006-02) Choi, Dong W.; Zea, Corbin J.; Do, Young S.; Semrau, Jeremy D.; Antholine, William E.; Hargrove, Mark S.; Pohl, Nicola L.; Boyd, Eric S.; Geesey, Gill G.; Hartsel, Scott C.; Shafe, Peter H.; McEllistrem, Marcus T.; Kisting, Clint J.; Campbell, Damon; Rao, Vinay; de la Mora, Arlene; DiSpirito, Alan A.To examine the potential role of methanobactin (mb) as the extracellular component of a copper acquisition system in Methylosinus trichosporium OB3b, the metal binding properties of mb were examined. Spectral (UV-visible, fluorescence, and circular dichroism), kinetic, and thermodynamic data suggested copper coordination changes at different Cu(II):mb ratios. Mb appeared to initially bind Cu(II) as a homodimer with a comparatively high copper affinity at Cu(II):mb ratios below 0.2, with a binding constant (K) greater than that of EDTA (log K ) 18.8) and an approximate ΔG° of -47 kcal/mol. At Cu(II):mb ratios between 0.2 and 0.45, the K dropped to (2.6 ± 0.46) 108 with a ΔG° of -11.46 kcal/mol followed by another K of (1.40 ± 0.21) 106 and a ΔG° of -8.38 kcal/mol at Cu(II):mb ratios of 0.45-0.85. The kinetic and spectral changes also suggested Cu(II) was initially coordinated to the 4-thiocarbonyl-5-hydroxy imidazolate (THI) and possibly Tyr, followed by reduction to Cu(I), and then coordination of Cu(I) to 4-hydroxy-5-thiocarbonyl imidazolate (HTI) resulting in the final coordination of Cu(I) by THI and HTI. The rate constant (kobsI) of binding of Cu(II) to THI exceeded that of the stopped flow apparatus that was used, i.e., >640 s-1, whereas the coordination of copper to HTI showed a 6-8 ms lag time followed by a kobsII of 121 ± 9 s-1. Mb also solubilized and bound Cu(I) with a kobsI to THI of >640 s-1, but with a slower rate constant to HTI (kobsII ) 8.27 ± 0.16 s-1), and appeared to initially bind Cu(I) as a monomer.Item Mineralogy influences structure and diversity of bacterial communities associated with geological substrata in a pristine aquifer(2007-03) Boyd, Eric S.; Cummings, David E.; Geesey, Gill G.Our understanding of mineralogical influences on subsurface microbial community structure and diversity has been difficult to assess due to difficulties in isolating this variable from others in the subsurface environment. In this study, biofilm coupons were used to isolate specific geological substrata from the surrounding geological matrix during colonization by microorganisms suspended in the surrounding groundwater for an 8-week period. Upon retrieval, the structure and diversity of the microbial community associated with each type of substratum was evaluated using 16S rDNA-based terminal-restriction fragment length polymorphism (T-RFLP). Phylogenetic affiliations of the populations associated with each type of substratum were established based on sequence analysis of near full-length 16S rDNA obtained through construction of a clone library. Hematite, quartz, and saprolite each harbored a community dominated by members of the division Proteobacteria (>67% of community). However, the different substrata selected for different subdivisions of bacteria within the Proteobacteria. After accounting for the influence exerted by substratum type on recovery of DNA from the attached populations, both phylogenetic data and Jaccard and Bray-Curtis similarity indices derived from terminal-restriction fragment (T-RF) profiles suggested a strong mineralogical influence on the structure and composition of the solid phase-associated community. The results suggest that mineralogical heterogeneity influences microbial community structure and diversity in pristine aquifers.Item Identification and characterization of a novel member of the radical AdoMet enzyme superfamily and implications for the biosynthesis of the Hmd hydrogenase active site cofactor(2009-11) McGlynn, Shawn E.; Boyd, Eric S.; Shepard, Eric M.; Lange, Rachel K.; Gerlach, Robin; Broderick, Joan B.; Peters, John W.The genetic context, phylogeny, and biochemistry of a gene flanking the H2-forming methylene-H4-methanopterin dehydrogenase gene (hmdA), here designated hmdB, indicate that it is a new member of the radical S-adenosylmethionine enzyme superfamily. In contrast to the characteristic CX3CX2C or CX2CX4C motif defining this family, HmdB contains a unique CX5CX2C motif.Item Diversity, abundance, and potential activity of nitrifying and nitrate-reducing microbial assemblages in a subglacial ecosystem(2011-05) Boyd, Eric S.; Lange, Rachel K.; Mitchell, Andrew C.; Havig, Jeff R.; Lafreniere, M. J.; Shock, Everett L.; Peters, John W.; Skidmore, Mark L.Subglacial sediments sampled from beneath Robertson Glacier (RG), Alberta, Canada, were shown to harbor diverse assemblages of potential nitrifiers, nitrate reducers, and diazotrophs, as assessed by amoA, narG, and nifH gene biomarker diversity. Although archaeal amoA genes were detected, they were less abundant and less diverse than bacterial amoA, suggesting that bacteria are the predominant nitrifiers in RG sediments. Maximum nitrification and nitrate reduction rates in microcosms incubated at 4°C were 280 and 18.5 nmol of N per g of dry weight sediment per day, respectively, indicating the potential for these processes to occur in situ. Geochemical analyses of subglacial sediment pore waters and bulk subglacial meltwaters revealed low concentrations of inorganic and organic nitrogen compounds. These data, when coupled with a C/N atomic ratio of dissolved organic matter in subglacial pore waters of ∼210, indicate that the sediment communities are N limited. This may reflect the combined biological activities of organic N mineralization, nitrification, and nitrate reduction. Despite evidence of N limitation and the detection of nifH, we were unable to detect biological nitrogen fixation activity in subglacial sediments. Collectively, the results presented here suggest a role for nitrification and nitrate reduction in sustaining microbial life in subglacial environments. Considering that ice currently covers 11% of the terrestrial landmass and has covered significantly greater portions of Earth at times in the past, the demonstration of nitrification and nitrate reduction in subglacial environments furthers our understanding of the potential for these environments to contribute to global biogeochemical cycles on glacial-interglacial timescales.Item Influence of bedrock mineral composition on microbial diversity in a subglacial environment(2013-06) Mitchell, Andrew C.; Lafrenière, M. J.; Skidmore, Mark L.; Boyd, Eric S.Microorganisms in subglacial environments drive the chemical weathering of bedrock; however, the influence of bedrock mineralogy on the composition and activity of microbial assemblages in such environments is poorly understood. Here, using a combination of in situ mineral incubation and DNA fingerprinting techniques, we demonstrate that pyrite is the dominant mineralogical control on subglacial bacterial community structure and composition. In addition, we show that the abundance of Fe in the incubated minerals influences the development of mineral-associated biomass. The ubiquitous nature of pyrite in many common bedrock types and high SO42– concentrations in most glacial meltwaters suggest that pyrite may be a dominant lithogenic control on microbial communities in many subglacial systems. Mineral-based energy may therefore serve a fundamental role in sustaining subglacial microbial populations and enabling their persistence over glacial-interglacial time scales.Item Hydrogenase activity of mineral-associated and suspended populations of Desulfovibrio desulfuricans Essex 6(2014-02) Reardon, Catherine L.; Magnuson, Timothy S.; Boyd, Eric S.; Leavitt, W. D.; Reed, D. W.; Geesey, Gill G.The interactions between sulfate-reducing microorganisms and iron oxides influence a number of important redox-sensitive biogeochemical processes including the formation of iron sulfides. Enzymes, such as hydrogenase which catalyze the reversible oxidation of molecular hydrogen, are known to mediate electron transfer to metals and may contribute to the formation and speciation of ferrous sulfides formed at the cell–mineral interface. In the present study, we compared the whole cell hydrogenase activity of Desulfovibrio desulfuricans strain Essex 6 growing as biofilms on hematite (hematite-associated) or as suspended populations using different metabolic pathways. Hematite-associated cells exhibited significantly greater hydrogenase activity than suspended populations during sulfate respiration but not during pyruvate fermentation. The enhanced activity of the hematite-associated, sulfate-grown cells appears to be dependent on iron availability rather than a general response to surface attachment since the activity of glass-associated cells did not differ from that of suspended populations. Hydrogenase activity of pyruvate-fermenting cells was stimulated by addition of iron as soluble Fe(II)Cl2 and, in the absence of added iron, both sulfate-reducing and pyruvate-fermenting cells displayed similar rates of hydrogenase activity. These data suggest that iron exerts a stronger influence on whole cell hydrogenase activity than either metabolic pathway or mode of growth. The location of hydrogenase to the cell envelope and the enhanced activity at the hematite surface in sulfate-reducing cells may influence the redox conditions that control the species of iron sulfides on the mineral surface.