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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 487 records · Page 27
All sourcesNASA NTRSDOE OSTI
DOE OSTI2018

Experimental tests on commercial Sweet Product Residue (SPR) as a suitable feed for anaerobic bioenergy (H2 + CH4) production

Highlights: • Post-sale refuses was tested as feed for two-step-Anaerobic Digestion (TSAD). • Extrusion was assessed at pilot plant scale process to remove the packaging. • NaOH Pretreatment was evaluated in order to increase the refuses digestibility. • H{sub 2} + CH{sub 4} production via TSAD process was experimentally evaluated. • Energy Sustainability Index were used to evaluate the process: pretreatment + TSAD. - Abstract: Food stores can find themselves in the position of having to dispose of different types of products, such as snacks, confectionery, prepackaged food, drinks and others, Sweet Product Residue (SPR), which presents a great opportunity to produce energy through Anaerobic Digestion (AD), due to its high sugar, carbohidrate and fat contents. In order to valorise SPR, this paper takes into consideration the all necessary treatments; owing the fact that the refuses are constitute by an organic part and packaging (plastic, paper and their combinations), a pretreatment able to remove the latter is necessary. SPR refuse was initially subjected to novel pretreatment approach: extrusion at 200 atm to remove the packaging, and a Basic Pre-treatment (BP) then tested through a Two-Stage AD (TSAD) process, for H{sub 2} and CH{sub 4} productions. The experimental results were analysed considering three parameters: Efficiency (ξ), which takes into account the quantity of the energy produced as hydrogen plus methane that the bioreaction is able to extract; Efficacy (η), which takes into account the efficiency of the actual test, compared with that obtained from a reference test carried out with glucose; Energy Sustainability Index (ESI), which takes into account the total amount of energy produced as H{sub 2} plus CH{sub 4}, and the amount of energy consumed to pre-treat the refuse. The effectiveness of the extrusion process in removing the packaging was very high: about 80% of the organic part present in the SPR was recovered. The obtained results have pointed out that SPR is suitable for energy valorization process: ξ = 50%, η = 0.67 and ESI = 24.4, without the need of basic pre-treatment.

09 BIOMASS FUELS↗
DOE OSTI2012

Lab-Scale Demonstration of Plutonium Purification by Anion Exchange, Plutonium(IV) Oxalate Precipitation, and Calcination to Plutonium Oxide to Support the MOX Feed Mission

H-Canyon and HB-Line are tasked with the production of PuO 2 from a feed of plutonium metal. The PuO 2 will provide feed material for the MOX Fuel Fabrication Facility. After dissolution of the Pu metal in H-Canyon, the solution will be transferred to HB-Line for purification by anion exchange. Subsequent unit operations include Pu(IV) oxalate precipitation, filtration and calcination to form PuO 2 . This report details the results from SRNL anion exchange, precipitation, filtration, calcination, and characterization tests, as requested by HB-Line1 and described in the task plan. This study involved an 80-g batch of Pu and employed test conditions prototypical of HB-Line conditions, wherever feasible. In addition, this study integrated lessons learned from earlier anion exchange and precipitation and calcination studies. H-Area Engineering selected direct strike Pu(IV) oxalate precipitation to produce a more dense PuO 2 product than expected from Pu(III) oxalate precipitation. One benefit of the Pu(IV) approach is that it eliminates the need for reduction by ascorbic acid. The proposed HB-Line precipitation process involves a digestion time of 5 minutes after the time (44 min) required for oxalic acid addition. These were the conditions during HB-line production of neptunium oxide (NpO 2 ). In addition, a series of small Pu(IV) oxalate precipitation tests with different digestion times were conducted to better understand the effect of digestion time on particle size, filtration efficiency and other factors. To test the recommended process conditions, researchers performed two nearly-identical larger-scale precipitation and calcination tests. The calcined batches of PuO 2 were characterized for density, specific surface area (SSA), particle size, moisture content, and impurities. Because the 3013 Standard requires that the calcination (or stabilization) process eliminate organics, characterization of PuO 2 batches monitored the presence of oxalate by thermogravimetric analysis-mass spectrometry (TGA-MS). To use the TGA-MS for carbon or oxalate content, some method development will be required. However, the TGA-MS is already used for moisture measurements. Therefore, SRNL initiated method development for the TGA-MS to allow quantification of oxalate or total carbon. That work continues at this time and is not yet ready for use in this study. However, the collected test data can be reviewed later as those analysis tools are available.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗
DOE OSTI2021

Phenylpropanoid Biosynthesis Gene Expression Precedes Lignin Accumulation During Shoot Development in Lowland and Upland Switchgrass Genotypes

Efficient conversion of lignocellulosic biomass into biofuels is influenced by biomass composition and structure. Lignin and other cell wall phenylpropanoids, such as para -coumaric acid ( p CA) and ferulic acid (FA), reduce cell wall sugar accessibility and hamper biochemical fuel production. Toward identifying the timing and key parameters of cell wall recalcitrance across different switchgrass genotypes, this study measured cell wall composition and lignin biosynthesis gene expression in three switchgrass genotypes, A4 and AP13, representing the lowland ecotype, and VS16, representing the upland ecotype, at three developmental stages [Vegetative 3 (V3), Elongation 4 (E4), and Reproductive 3 (R3)] and three segments (S1–S3) of the E4 stage under greenhouse conditions. A decrease in cell wall digestibility and an increase in phenylpropanoids occur across development. Compared with AP13 and A4, VS16 has significantly less lignin and greater cell wall digestibility at the V3 and E4 stages; however, differences among genotypes diminish by the R3 stage. Gini correlation analysis across all genotypes revealed that lignin and p CA, but also pectin monosaccharide components, show the greatest negative correlations with digestibility. Lignin and p CA accumulation is delayed compared with expression of phenylpropanoid biosynthesis genes, while FA accumulation coincides with expression of these genes. The different cell wall component accumulation profiles and gene expression correlations may have implications for system biology approaches to identify additional gene products with cell wall component synthesis and regulation functions.

09 BIOMASS FUELS↗
DOE OSTI2015

Presence of Siloxanes in the Biogas of a Wastewater Treatment Plant Separation in Condensates and Influence of the Dose of Iron Chloride on its Elimination

The siloxanes present in the biogas produced during anaerobic digestion damage the mechanism of cogeneration equipment and, consequently, negatively affect the energy valorization process. For this reason, the detection and elimination of these silicon-derived chemical compounds are a priority in the management of cogeneration facilities. In this regard, the objectives of this paper are, firstly, to characterize the siloxanes in the biogas and, secondly, to qualitatively evaluate the influence of the dose of iron chloride on its elimination. The research was performed at the Rincón de León Wastewater Treatment Plant (Alicante, Spain). The outflow biogas of the digesters and of the pressurized gasometers was sampled and analyzed. The results obtained made it possible to demonstrate, firstly, the absence of linear siloxanes and that, of the cyclic siloxanes, the predominant type was decamethylcyclopentasiloxane, and, secondly, that the addition of iron chloride in the digesters significantly reduces the siloxane content in the biogas. Additionally, it was demonstrated that the process of compression of the biogas, with the elimination of condensates, also produces significant reductions in the concentration of siloxanes in the biogas.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2021

Qualitative and quantitative profiles of jejunal oligosaccharides and cecal short–chain fatty acids in broiler chickens receiving different dietary levels of fiber, protein and exogenous enzymes

Dietary supplemental carbohydrases are able to degrade non-starch polysaccharides and generate oligosaccharides in the gastrointestinal tract. This study was conducted to investigate the influence of dietary fiber and protein levels on growth performance, nutrient utilization, digesta oligosaccharides profile and cecal short-chain fatty acid (SCFA) profile in broilers receiving diets supplemented with xylanase or protease individually or in combination. Enzyme supplementation had no effect on growth performance. There was significant (P < 0.05) fiber × protein × xylanase interaction for ileal nitrogen digestibility and significant (P < 0.01) protein × xylanase × protease interaction for nitrogen-corrected apparent metabolizable energy. Birds fed high-fiber diets had higher (P < 0.05) jejunal oligosaccharides and cecal SCFA concentrations. Xylanase and protease combination produced the greatest pentose (Pent) levels in low fiber–adequate protein diets but lowest levels in highfiber–low protein diets. There was significant (P < 0.05) fiber × xylanase × protease interaction explained by the digesta concentrations of (Pent) 3 , (Pent) 4 and (Pent) 5 being greatest (P < 0.5) in protease-only supplemented high-fiber diets but lowest in protease-only supplemented low-fiber diets. Furthermore, these results suggest that, of all the factors investigated, dietary fiber level had the greatest effect on modulating digesta concentration of oligosaccharides and cecal SCFA. Evidence points to the fact that there is considerable capacity for generating pentose oligosaccharides in the digestive tract of broilers receiving diets rich in fibrous feedstuffs, and that this may have a beneficial effect on microbial profile in the digestive tract.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2015

Dissolution of aerosol particles collected from nuclear facility plutonium production process

Here, a simple, robust analytical chemistry method has been developed to dissolve plutonium containing particles in a complex matrix. The aerosol particles collected on Marple cascade impactor substrates were shown to be dissolved completely with an acid mixture of 12 M HNO 3 and 0.1 M HF. A pressurized closed vessel acid digestion technique was utilized to heat the samples at 130 °C for 16 h to facilitate the digestion. The dissolution efficiency for plutonium particles was 99 %. The resulting particle digestate solution was suitable for trace elemental analysis and isotope composition determination, as well as radiochemistry measurements.

38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEA↗
DOE OSTI2018

Structural and dynamical characteristics of tropomyosin epitopes as the major allergens in shrimp

Highlights: • The epitope region in tropomyosin was more stable than the non-epitope region. • The difference between epitope and non-epitope regions was largest at low pHs. • Lower content of Ala cluster is involved in higher stability of the epitope region. Ingestion of marine invertebrates often causes food allergy, where the major allergens have been reported to be derived from tropomyosin (TM). Intact or the digestive fragments of food allergens generally show resistance to digestion, which is usually attributable to the structural stability (or rigidity). The difference in the structural and dynamical characteristics between the epitope and the non-epitope regions in TM has not yet been well understood. In the present study, molecular dynamics simulation was performed at constant pHs for shrimp TM. By analyzing the main-chain dihedral angle fluctuations and local α-helix contents, we found that the epitope regions are more stable than the non-epitope counterparts, providing a possible physical reason for the resistance to digestion in the epitopes regions. The difference of the structural stability between the epitope and the non-epitope regions was largest at low pHs, even though pH dependence of the structural stability in itself was not significant in both regions. The lower content of the Ala cluster in the epitope region is considered to cause the higher stability of the epitope region.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Yap1/Taz are essential for the liver development in zebrafish

Highlights: • Taz is significantly up-regulated in zebrafish yap1 mutants. • yap1{sup −/−}; taz{sup +/−} embryos are defective in digestive organs development. • Mutations in yap1{sup −/−}; taz{sup +/−} impair cell proliferation in liver and movement of LPM. • Taz can substituteYap1's function in liver development in zebrafish. Hippo pathway regulates cell proliferation and differentiation. Yes-associated protein (Yap) and transcriptional coactivator with PDZ-binding motif (Taz) are effectors of Hippo pathway. The function of Yap/Taz in embryonic liver development has yet to be reported. Here yap1 and taz were found expressed in liver and other digestive organs in zebrafish embryos, and knockout of yap1 or taz did not lead to visible defects during embryogenesis. Interestingly, Taz was significantly increased in yap1 mutants, which may account for their normal development, albeit losing Yap1. However, yap1{sup −/−}; taz{sup +/-} embryos exhibited smaller digestive organs, and more than half of them showed bilateral livers and pancreas and non-looped intestines. Further analysis revealed that the disrupted gene function in yap1{sup −/−}; taz{sup +/-} embryos did not disturb liver bud formation, but instead impaired cell proliferation in liver and movement of the neighboring lateral plate mesoderm (LPM). Overexpression of wild type yap1 or taz could rescue the defective liver phenotypes in yap1{sup −/−}; taz{sup +/-} embryos, indicating that Yap1 cooperate with Taz to regulate the liver development. In addition, Yap1 was found to function in a Tead-dependent manner in the liver development. Our results suggest that Yap1/Taz regulate LPM movement and promote cell proliferation to ensure proper liver development in zebrafish.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2025

Evaluating size exclusion chromatography for nucleic acid removal in Klebsiella pneumoniae cell surface polysaccharide purification

Cell surface-associated polysaccharides in Klebsiella are major virulence determinants and crucial targets for developing vaccines. Traditionally, the purification of these cell surface polysaccharides from Klebsiella pneumoniae involves a multi-step process comprising phenol extraction, nuclease digestion, ultracentrifugation, and repeated ethanol extractions. In this study, we evaluated size exclusion chromatography for effectively eliminating nucleic acid contamination while purifying high molecular weight cell surface-associated polysaccharides. Post-initial extraction, the nucleic acid content remains significantly elevated, and kinetic analysis reveals that DNase I and RNase A digestion is neither economically viable nor effective for removing these contaminants. Employing an appropriate size exclusion resin removes over 99 % of nucleic acid contamination, as confirmed by nucleic acid content analysis and agarose gel electrophoresis. Purity and structural analysis using 1H 1D-NMR and 2D-NMR demonstrate that the cell surface-associated polysaccharide purified with this study is highly homogeneous and identified as antigenic O-polysaccharide. This approach streamlines the purification process by removing the need for nuclease digestion and additional ethanol precipitation steps.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Protein-based forensic identification using genetically variant peptides in human bone

Bone tissue contains organic material that is useful for forensic investigations and may contain preserved endogenous protein that can persist in the environment for extended periods of time over a range of conditions. Single amino acid polymorphisms in these proteins reflect genetic information since they result from non-synonymous single nucleotide polymorphisms (SNPs) in DNA. Detection of genetically variant peptides (GVPs) — those peptides that contain amino acid polymorphisms — in digests of bone proteins allows for the corresponding SNP alleles to be inferred. Resulting genetic profiles can be used to calculate statistical measures of association between a bone sample and an individual. In this study proteomic analysis on rib cortical bone samples from 10 recently deceased individuals demonstrates this concept. A straight-forward acidic demineralization protocol yielded proteins that were digested with trypsin. Tryptic digests were analyzed by liquid chromatography mass spectrometry. A total of 1736 different proteins were identified across all resulting datasets. On average, individual samples contained 454 ± 121 ($\bar{x}$ ± σ) proteins. Thirty-five genetically variant peptides were identified from 15 observed proteins. Overall, 134 SNP inferences were made based on proteomically detected GVPs, which were confirmed by sequencing of subject DNA. Inferred individual SNP genetic profiles ranged in random match probability (RMP) from 1/6 to 1/42,472 when calculated with European population frequencies in the 1000 Genomes Project, Phase 3. Similarly, RMPs based on African population frequencies were calculated for each SNP genetic profile and likelihood ratios (LR) were obtained by dividing each European RMP by the corresponding African RMP. In conclusion, resulting LR values ranged from 1.4 to 825 with a median value of 16. GVP markers offer a basis for the identification of compromised skeletal remains independent of the presence of DNA template.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2016

Synthesis and catalytic activity of the metastable phase of gold phosphide

Recently, transition metal phosphides have found new applications as catalysts for the hydrogen evolution reaction that has generated an impetus to synthesize these materials at the nanoscale. In this work, Au{sub 2}P{sub 3} was synthesized utilizing the high temperature decomposition of tri-n-octylphosphine as a source of elemental phosphorous. Gold nanorods were used as morphological templates with the aim of controlling the shape and size of the resulting gold phosphide particles. We demonstrate that the surface capping ligand of the gold nanoparticle precursors can influence the purity and extent to which the gold phosphide phase will form. Gold nanorods functionalized with 1-dodecanethiol undergo digestive ripening to produce discrete spherical particles that exhibit reduced reactivity towards phosphorous, resulting in low yields of the gold phosphide. In contrast, gold phosphide was obtained as a phase pure product when cetyltrimethylammonium bromide functionalized gold nanorods are used instead. The Au{sub 2}P{sub 3} nanoparticles exhibited higher activity than polycrystalline gold towards the hydrogen evolution reaction. - Graphical abstract: Au{sub 2}P{sub 3} was synthesized utilizing the high temperature decomposition of tri-n-octylphosphine as a source of elemental phosphorous and gold nanoparticles as reactants. We demonstrate that the surface capping ligand of the gold nanoparticle precursors influence the purity and extent to which the Au{sub 2}P{sub 3} phase will form. Gold nanorods functionalized with 1-dodecanethiol undergo digestive ripening to produce discrete spherical particles that exhibit reduced reactivity towards phosphorous, resulting in low yields of the gold phosphide. In contrast, gold phosphide was obtained as a phase pure product when cetyltrimethylammonium bromide functionalized gold nanoparticles are used instead. The Au{sub 2}P{sub 3} nanoparticles exhibited higher activity than polycrystalline gold towards the hydrogen evolution reaction. - Highlights: • The surface chemistry of gold affects the synthetic yields of Au{sub 2}P{sub 3}. • Imaging of Au{sub 2}P{sub 3} with transmission electron microscopy results in decomposition. • Au{sub 2}P{sub 3} nanoparticles exhibit activity towards the hydrogen evolution reaction.

36 MATERIALS SCIENCE↗
DOE OSTI2014

Evaluation of gold and silver leaching from printed circuit board of cellphones

Highlights: • Printed circuit boards (PCB) of mobile phones have large amounts of metals with high economic value such as gold and silver. • Dissolution of gold was done with a cyanide-based reagent and silver with nitric acid. • Leaching of PCB with Na{sub 2}S{sub 2}O{sub 3} and (NH{sub 4}){sub 2}S{sub 2}O{sub 3} to examine the feasibility of using these reagents was done. - Abstract: Electronic waste has been increasing proportionally with the technology. So, nowadays, it is necessary to consider the useful life, recycling, and final disposal of these equipment. Metals, such as Au, Ag, Cu, Sn and Ni can be found in the printed circuit boards (PCB). According to this, the aims of this work is to characterize the PCBs of mobile phones with aqua regia; obtaining “reference” values of leaching, to gold and silver, with cyanide and nitric acid, respectively; and study the process of leaching of these metals in alternative leaching with sodium thiosulfate and ammonium thiosulfate. The metals were characterized by digesting the sample with aqua regia for 1 and 2 h at 60 °C and 80 °C. The leaching of Au with a commercial reagent (cyanide) and the Ag with HNO{sub 3}were made. The leaching of Au and Ag with alternative reagents: Na{sub 2}S{sub 2}O{sub 3,} and (NH{sub 4}){sub 2}S{sub 2}O{sub 3} in 0.1 M concentration with the addition of CuSO{sub 4}, NH{sub 4}OH, and H{sub 2}O{sub 2}, was also studied. The results show that the digestion with aqua regia was efficient to characterize the metals present in the PCBs of mobile phones. However, the best method to solubilize silver was by digesting the sample with nitric acid. The leaching process using sodium thiosulfate was more efficient when an additional concentration of 0.015 and 0.030 M of the CuSO{sub 4} was added.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2018

Environmental assessment of alternative treatment schemes for energy and nutrient recovery from livestock manure

Highlights: • Energy and nutrient recovery from manure was environmentally evaluated. • Direct digestate application is not always feasible. • AD+centrifugation+N-removal was the most environmentally favourable scenario. • N-removal through HRAS+PNA considerably improved the environmental performance. • UF membrane+acidification were needed to improve struvite quantity and quality. - Abstract: The application of livestock manure on agricultural land is being restricted due to its significant content of phosphorus (P) and nitrogen (N), leading to eutrophication. At the same time, the growing demand for N and P mineral fertilizers is increasing their production costs and causing the depletion of natural phosphate rock deposits. In the present work, seven technologically feasible treatment schemes for energy (biogas) and nutrient recovery (e.g., struvite precipitation) and/or removal (e.g., partial nitritation/anammox) were evaluated from an environmental perspective. In general, while approaches based solely on energy recovery and use of digestate as fertilizer are commonly limited by community regulations, strategies pursuing the generation of high-quality struvite are not environmentally sound alternatives. In contrast, schemes that include further solid/liquid separation of the digestate improved the environmental profile, and their combination with an additional N-removal stage would lead to the most environmental-friendly framework. However, the preferred scenario was identified to be highly dependent on the particular conditions of each site, integrating environmental, social and economic criteria.

09 BIOMASS FUELS↗
DOE OSTI2018

Carrier-Assisted Single-Tube Processing Approach for Targeted Proteomics Analysis of Low Numbers of Mammalian Cells

Heterogeneity in composition is inherent in all cell populations, even those containing a single cell type. Single-cell proteomics characterization of cell heterogeneity is currently achieved by antibody-based technologies, which are limited by the availability of highquality antibodies. In this paper we report a simple, easily implemented, mass spectrometry (MS)- based targeted proteomics approach, termed cLC-SRM (carrier-assisted liquid chromatography coupled to selected reaction monitoring), for reliable multiplexed quantification of proteins in low numbers of mammalian cells. We combine a new single-tube digestion protocol to process low numbers of cells with minimal loss together with sensitive LC-SRM for protein quantification. This single-tube protocol builds upon trifluoroethanol digestion and further minimizes sample losses by tube pretreatment and the addition of carrier proteins. We also optimized the denaturing temperature and trypsin concentration to significantly improve digestion efficiency. cLC-SRM was demonstrated to have sufficient sensitivity for reproducible detection of most epidermal growth factor receptor (EGFR) pathway proteins expressed at levels =30 000 and =3000 copies per cell for 10 and 100 mammalian cells, respectively. Thus, cLC-SRM enables reliable quantification of low to moderately abundant proteins in less than 100 cells and could be broadly useful for multiplexed quantification of important proteins in small subpopulations of cells or in size-limited clinical samples. Further improvements of this method could eventually enable targeted single-cell proteomics when combined with either SRM or other emerging ultrasensitive MS detection.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2019

Strategies to Valorize the Hydrothermal Liquefaction-Derived Aqueous Phase into Fuels and Chemicals

Valorization options for the aqueous fraction produced by the hydrothermal liquefaction of terrestrial feedstocks were developed. The processes focused on carboxylic acids, which are abundant in the aqueous fraction. A clean up process for the aqueous phase based on activated carbon absorption followed by liquid-liquid extraction was demonstrated that significantly reduces color bodies and inorganic contaminants while simultaneously concentrating desirable organic constituents. Conversion of the cleaned aqueous phase was then demonstrated for both a ketonization reaction and steam reforming. Condensed phase ketonization over LaxZryOz catalyst was found to exhibit both excellent hydrothermal stability and ketonization activity. Necessity for feedstock cleanup was established and stable condensed phase ketonization operation was demonstrated for ~100 hours with HTL-process derived material. Steam reforming was also demonstrated by employing a dual-bed catalyst configuration whereby carboxylic acids first undergo ketonization over CeO2 in the vapor phase followed by steam reforming of the ketone intermediates over Co/CeO2. The dual-bed approach reduced coke deposition and thus catalyst stability improved. Techno-economic analysis was used to assess economic viability for producing fuel from the bio-oil fraction and either propene (formed via hydrogenation and dehydration of acetone, the primary ketonization product) or hydrogen from the aqueous stream. The ketonization and steam reforming cases were separately modeled and benchmarked to a process utilizing anaerobic digestion. Resulting minimum fuel selling prices of $3.7, $4.2, and $4.3/GGE were obtained for the production of olefins via ketonization, reduction and dehydration; steam reforming; and anaerobic digestion processes, respectively. Both the steam reforming and anaerobic digestion processes benefited by avoiding the need for natural gas by producing the hydrogen on-site required for biocrude hydrotreatment. However, the condensed ketonization process had the lowest minimum fuel selling price because of the co-product olefins generated for sale.

Biomass, hydrothermal liquefaction, techno economi↗
DOE OSTI2013

Sources of Technical Variability in Quantitative LC-MS Proteomics: Human Brain Tissue Sample Analysis.

To design a robust quantitative proteomics study, an understanding of both the inherent heterogeneity of the biological samples being studied as well as the technical variability of the proteomics methods and platform is needed. Additionally, accurately identifying the technical steps associated with the largest variability would provide valuable information for the improvement and design of future processing pipelines. We present an experimental strategy that allows for a detailed examination of the variability of the quantitative LC-MS proteomics measurements. By replicating analyses at different stages of processing, various technical components can be estimated and their individual contribution to technical variability can be dissected. This design can be easily adapted to other quantitative proteomics pipelines. Herein, we applied this methodology to our label-free workflow for the processing of human brain tissue. For this application, the pipeline was divided into four critical components: Tissue dissection and homogenization (extraction), protein denaturation followed by trypsin digestion and SPE clean-up (digestion), short-term run-to-run instrumental response fluctuation (instrumental variance), and long-term drift of the quantitative response of the LC-MS/MS platform over the 2 week period of continuous analysis (instrumental stability). From this analysis, we found the following contributions to variability: extraction (72%) >> instrumental variance (16%) > instrumental stability (8.4%) > digestion (3.1%). Furthermore, the stability of the platform and its’ suitability for discovery proteomics studies is demonstrated.

Environmental Molecular Sciences Laboratory↗
DOE OSTI2019

Cellulose hydrolysis by Clostridium thermocellum is agnostic to substrate structural properties in contrast to fungal cellulases

The native recalcitrance of lignocellulosic biomass hinders its effective deconstruction for biological conversion to fuel ethanol. However, once cellulose is physically available to enzymes/microbes, i.e., macro-accessible, cellulose micro-accessibility, i.e., the accessibility as influenced by cellulose properties, further affects cellulose conversion. Here, we performed a comparative study of the effect of cellulose micro-accessibility on cellulose conversion by two biological approaches of potential commercial interest: consolidated bioprocessing (CBP) using Clostridium thermocellum and cell-free saccharification mediated by fungal enzymes. Commercially available cellulosic substrates, Avicel® PH-101, Sigmacell Cellulose Type 50, cotton linters, Whatman™ 1 milled filter paper, and α-cellulose were employed to constitute different cellulose micro-accessibilities. Physiochemical characterization was performed on these substrates to determine key morphological and chemical differences. Biological conversion of these substrates showed that C. was unaffected overall by cellulose structural properties, i.e., micro-accessibility, and achieved similar solids solubilization and metabolite production from these structurally different materials. However, fungal enzymes digested these substrates to different extents. Specifically, glucan conversion of these substrates diminished in the following order: milled filter paper > Avicel > Sigmacell and α-cellulose > cotton linters. Here, we propose that C. thermocellum digestion of lignocellulosic biomass is primarily controlled by the physical availability of cellulose in the lignocellulosic matrix and largely unaffected by cellulose properties once cellulose is made macro-accessible. Finally, in contrast, fungal enzymes require cellulose to be physically accessible, i.e., macro-accessible, as well as have properties amenable to digestion, i.e., micro-accessible.

09 BIOMASS FUELS↗