FY2010 - Oak Ridge National Laboratory

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Director’s R&D Fund— Systems Biology and the Environment Results and Accomplishments We have identified current wood harvest and storage as a proxy for woody sequestration potential constrained, to first approximation, by land availability, forest productivity, cost of extraction, etc. Estimates of current global wood harvest are on order 1–1.5 Gt carbon per year. Approximately 30% of that estimate is waste burned or left to decay at harvest. Additional carbon is lost during conversion to wood products, with only 20–25% of delivered wood harvest sequestered in long-lived products or landfills. We estimate accordingly that current global wood product sequestration is on order 0.1–0.2 Gt carbon per year. Allowing for realistic intensification of production, efficiency, and management, potentials for woody biosequestration on order 0.1–1.0 Gt carbon per year are perhaps likely. In a complementary top-down analysis we find that an unconstrained theoretical global potential of 10.0 Gt carbon per year is reduced to 6 Gt carbon per year after accounting for agriculture and other land-use constraints. This potential is further reduced to 3.5 Gt carbon per year when conservation of forestland for biodiversity and other ecosystem services is considered. The allowance for other uses of wood produced on forested land reduces the potential for carbon sequestration and emissions mitigation further to an upper estimate of perhaps 2.5 Gt carbon per year. The bottom-up and top-down analyses together suggest a realizable potential for woody biomass sequestration in the range of 0.1 to 3.0 Gt carbon per year. 05833 Harnessing Nitrogen and Sulfur Cycles to Develop Microbial Consortia for Consolidated Bioprocessing David Graham Project Description Anaerobic digesters use microorganisms to convert complex polymers into smaller organic compounds, including biofuels. While most studies focus on carbon flux in these digesters, understanding nitrogen and sulfur fluxes will be essential for maintaining microbial biocatalysts and processing fermentation wastes. After primary fermentation, both elements will be enriched in the residue, and subsequent combustion will produce NO x and SO 2 gases. These gas emissions are regulated, so we want to sequester and deplete their precursors from waste. This project will characterize nitrogen and sulfur flux in cellulolytic systems and develop biochemical tools for monitoring enzymatic activity in complex mixtures. Microbial cocultures of cellulose-degrading bacteria and nitrate- or sulfur-reducing microorganisms will alter the nitrogen and sulfur cycles and create evolutionarily stable, mutualistic consortia. When one member of these consortia can be genetically altered, we can modify the system to increase cellulose degradation rates, reduce nitrogen and sulfur residues, and produce novel, high-value bioproducts. Mission Relevance This project will demonstrate engineered systems of microbial co-cultures for consolidated cellulose bioprocessing. The results, reagents, and tools developed here will serve as preliminary data for future grant proposals in biofuel, carbon cycling, waste management, and bioprocessing areas. This project anticipates future funding initiatives from the DOE Office of Biological and Environmental Research (BER), as it addresses two primary goals: develop biofuels as energy resources and determine the relationships between climate change and terrestrial processes. These activities could also provide the basis for Environmental Protection Agency (EPA) or U.S. Department of Agriculture (USDA) funding to reduce greenhouse gas (GHG) emissions from bioprocesses and alter the nutritional content of fermentation residue for animal feed products. Microbiological and biochemical expertise developed in 104
Director’s R&D Fund— Systems Biology and the Environment this project will be deployed in a BioEnergy Science Center (BESC)–funded enzymological task during FY 2011. Follow-on funding is also being pursued through a University of Tennessee–led collaborative proposal to the <strong>National</strong> Science Foundation (NSF). Results and Accomplishments Since its inception during December 2009, this project has progressed quickly towards meeting the firstyear goals. Defined, minimal growth media have been developed for the cellulose and hemicellulosedegrading bacteria Caldicellulosiruptor bescii, Caldicellulosiruptor obsidiansis, and Clostridium thermocellum. Expensive and complex nutrient supplements were replaced with individual vitamins, based on genome-enabled metabolic predictions. Specific nitrogen and sulfur source requirements have been identified for these organisms, substantially reducing levels of those nutrients added to the growth media. We are currently using these minimal, defined media to develop co-cultures of compatible thermophilic microorganisms with enhanced substrate range and metabolic versatility. These platforms will be used to expedite metabolic engineering for biomass deconstruction and waste product reduction. Polyclonal antibodies specific to Caldicellulosiruptor cells have been developed and validated, along with high-throughput, microplate-based fluorescence assays of glycosidase enzymatic activity, to rapidly characterize mixed microbial communities. -Glucosidase, endoglucanase, and xylosidase activities can now be measured rapidly with high sensitivity. Additional high-throughput assays permit the rapid analysis of biomass in complex matrices (as total protein) and total alcohol production during fermentation. These methods are now being used to evaluate the effects of nitrogen and sulfur concentrations on pre-treated poplar fermentation. New studies of co-cultures are applying these analytical methods to rapidly assess changes in biomass deconstruction and ethanol production. Information Shared Graham, D. E. 2010. “A new role for coenzyme F 420 in aflatoxin reduction by soil mycobacteria.” Mol. Microbiol. 78, 533–536. 105
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NEUTRON SCIENCES ..................
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NATIONAL SECURITY SCIENCE AND TECHN
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05887 Controlling the Catalytic Pro
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Introduction INTRODUCTION The Labor
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Index of Project Numbers 05501 ....
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FY2010 - Oak Ridge National Laboratory

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