Sandia Mechanics Challenge: Experimental Testing for Calibration and Impact Testing of a Structure with a Thread Fastener Joint
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Biological engineering of microorganisms to produce value-added chemicals is a promising route to sustainable manufacturing. However, overproduction of metabolic intermediates at high titer, rate, and yield from inexpensive substrates is challenging in non-model systems where limited information is available regarding metabolic flux and its control in production conditions. Integrated multi-omic analyses of engineered strains offers an in-depth look at metabolites and proteins directly involved in growth and production of target and non-target bioproducts. Here we applied multi-omic analyses to overproduction of the polymer precursor 3-hydroxypropionic acid (3HP) in the filamentous fungus Aspergillus pseudoterreus. A synthetic pathway consisting of aspartate decarboxylase, beta-alanine pyruvate transaminase, and 3HP dehydrogenase was designed and built for A. pseudoterreus. Strains with single- and multi-copy integration events were isolated and multi-omics analysis consisting of intracellular and extracellular metabolomics and targeted and global proteomics was used to interrogate the strains in shake-flask and bioreactor conditions. Production of a variety of co-products (organic acids and glycerol) and oxidative degradation of 3HP were identified as metabolic pathways competing with 3HP production. Intracellular accumulation of nitrogen as 2,4-diaminobutanoate was identified as an off-target nitrogen sink that may also limit flux through the engineered 3HP pathway. Elimination of the high-expression oxidative 3HP degradation pathway by deletion of a putative malonate semialdehyde dehydrogenase improved the yield of 3HP by 3.4× after 10 days in shake-flask culture. This is the first report of 3HP production in a filamentous fungus amenable to industrial scale biomanufacturing of organic acids at high titer and low pH.
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46th International Technical Conference on Clean Energy, Clearwater, FL, August 1-4, 2022
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Location: INL Materials and Fuels Complex (MFC); Zero Power Physics Reactor (ZPPR) Facility Purpose: Provides infrastructure to support DOE authorized experiments requiring a Hazards Category 2 facility with an elevated security posture
A radiation shielding system is needed to protect personnel and equipment from harsh neutron and photon fluxes during: Reactor operations Post shut down operations Disassembly Decommissioning Examples of RSS driving requirements Radiological Controls Operating dose rate: the dose rate during normal reactor operations shall be <0.5 mrem/hr at the exterior wall of DOME containment Outdoor radiation limit: shielding shall provide the dose to a facility worker outside of DOME to <0.05 mrem/hr at 50 ft from the exterior wall of the DOME Temperature Reactors may have operating temperatures up to 800C The RSS shall be capable of removing up to 100 KW of heat during normal operating conditions the concrete structure has an upper temperature limit of 100°C
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