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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 1,027 records · Page 57
All sourcesNASA NTRSDOE OSTI
DOE OSTI2021

Supplemental Defatted Microalgae Nannochloropsis oceanica Affected Apparent Retention and Ileal Digestibility of Nutrients in a Corn-Soybean Meal Based Diet for Broiler Chickens

Objective: This study was to assess impacts of including 10% defatted microalgae (Nannochloropsis oceanica, 45% crude protein and 3.8% ether extract) into a corn-soybean meal basal diet for broiler chickens on apparent retention and ileal digestibility of nutrients. Procedures: Day-old hatchling Cornish Giant cockerels were divided into two groups (6 cages/group, 6 chicks/cage) and fed either the basal (control) or the microalgae-supplemented (microalgae) diet for 6 weeks. Starting week 3, the two experimental diets were mixed with 0.2% chromium oxide as an indigestible marker. Total excreta of individual cages were collected daily for 3 consecutive days during week 6. At the end of week 6, chicks were euthanized to collect ileal digesta from 1 chick/cage. Energy and concentrations of dry matter, crude protein, ether extract, minerals, and amino acids in digesta, excreta, and diets were assayed. Apparent retentions and ileal digestibilities were calculated using the direct collection and indirect indicator methods. Data were analyzed by the Student’s t-test and the significant level of difference was at p < 0.05. Results: The microalgae diet had elevated (4.2% and 8.1% by the two methods, p < 0.05) apparent retention of ether extract, but similar apparent retention and ileal digestibility of dry matter and apparent metabolizable energy, compared with the control diet. Supplemental defatted microalgae decreased (p < 0.05) the apparent retention and ileal digestibility of dietary crude protein by 13% and 13%. The supplementation also decreased (p < 0.05) ileal digestibilities and apparent retentions of multiple amino acids ranging from 0.6% to 26%. Apparent retentions of sodium and total phosphorus were decreased (p < 0.05) by 13% and 71%, respectively, by the microalgae inclusion compared with the control. Conclusion: The inclusion of 10% defatted N. oceanica in the corn-soybean meal basal diet for broiler chickens led moderate increases in apparent retention and ileal digestibility of ether extract, but decreased those of crude protein, amino acids, sodium, and phosphorus.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2022

N and P constrain C in ecosystems under climate change: Role of nutrient redistribution, accumulation, and stoichiometry

Abstract We use the Multiple Element Limitation (MEL) model to examine responses of 12 ecosystems to elevated carbon dioxide (CO 2 ), warming, and 20% decreases or increases in precipitation. Ecosystems respond synergistically to elevated CO 2 , warming, and decreased precipitation combined because higher water‐use efficiency with elevated CO 2 and higher fertility with warming compensate for responses to drought. Response to elevated CO 2 , warming, and increased precipitation combined is additive. We analyze changes in ecosystem carbon (C) based on four nitrogen (N) and four phosphorus (P) attribution factors: (1) changes in total ecosystem N and P, (2) changes in N and P distribution between vegetation and soil, (3) changes in vegetation C:N and C:P ratios, and (4) changes in soil C:N and C:P ratios. In the combined CO 2 and climate change simulations, all ecosystems gain C. The contributions of these four attribution factors to changes in ecosystem C storage varies among ecosystems because of differences in the initial distributions of N and P between vegetation and soil and the openness of the ecosystem N and P cycles. The net transfer of N and P from soil to vegetation dominates the C response of forests. For tundra and grasslands, the C gain is also associated with increased soil C:N and C:P. In ecosystems with symbiotic N fixation, C gains resulted from N accumulation. Because of differences in N versus P cycle openness and the distribution of organic matter between vegetation and soil, changes in the N and P attribution factors do not always parallel one another. Differences among ecosystems in C‐nutrient interactions and the amount of woody biomass interact to shape ecosystem C sequestration under simulated global change. We suggest that future studies quantify the openness of the N and P cycles and changes in the distribution of C, N, and P among ecosystem components, which currently limit understanding of nutrient effects on C sequestration and responses to elevated CO 2 and climate change.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2018

Optimization of signal-to-noise ratio in the in vivo{sup 31}P magnetic resonance spectra of the human brain

The main problem in {sup 31}P magnetic resonance spectroscopy is a low signal-to-noise ratio (SNR) of spectra acquired with clinical magnetic resonance imaging (MRI) scanners. Using spin-spin phosphorus-proton ({sup 31}P-{sup 1}H) decoupling and heteronuclear Overhauser effect and taking into account the effect of the longitudinal relaxation time T{sub 1} on the SNR, the method for localization and excitation of the region of interest (Image Selected in vivo Spectroscopy pulse sequence) was optimized to increase the SNR in the {sup 31}P magnetic resonance spectra of the human brain to ~50% without increasing signal acquisition time.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2018

Carbothermic Reduction Kinetics of Phosphorous Vaporization from Tri-calcium Phosphate (TCP) Under Microwave Rapid Heating With/Without the Presence of Fe{sub 3}O{sub 4}

The kinetics of vapor phase dephosphorization from tri-calcium phosphate (TCP) by carbothermic reduction was studied with and without the presence of Fe{sub 3}O{sub 4}. Microwave heating was utilized to obtain large variations in the heating rate (HR). In the reduction of TCP alone, the phosphorous removal fraction (RF; equal to ∆P{sub 2}O{sub 5}/P{sub 2}O{sub 5}{sup 0}, where ∆P{sub 2}O{sub 5} is the weight change and P{sub 2}O{sub 5}{sup 0} is the P{sub 2}O{sub 5} weight before heating) decreased as the HR increased. In other words, a shorter residence time at a high temperature resulted in a smaller reduction fraction of TCP. An apparently third-order reaction was postulated to account for the kinetics using a fitting simulation based on the additive law of the reaction progress. On the other hand, the phosphorous removal (dephosphorization) rate (RR; equal to ∆P{sub 2}O{sub 3}/t{sub MW}, where t{sub MW} is the microwave heating time period) increased as the HR increased above 1200 °C. The reduction ratio of Fe{sub 3}O{sub 4} above 1100 °C is higher than 97 pct regardless of the heating rate. The reduction of TCP in the presence of Fe{sub 3}O{sub 4} showed that RF increased slightly with increasing HR despite a shorter residence time at a high temperature. The RR also increased with the HR even though RF decreased to half of the values observed in the cases without Fe{sub 3}O{sub 4} for temperatures above 1200 °C. The practicality and optimal operation conditions of phosphorus vapor removal were discussed.

36 MATERIALS SCIENCE↗
DOE OSTI2018

Effect of Direct Reduced Iron (DRI) on Dephosphorization of Molten Steel by Electric Arc Furnace Slag

We investigated the effect of direct reduced iron (DRI) addition on dephosphorization of molten steel by electric arc furnace (EAF) slag at 1823 K (1550 °C). Various phenomena such as CO gas evolution and slagmaking by gangue oxides in DRI were experimentally observed at each reaction step. Thermodynamic behaviors of phosphorus, oxygen, and carbon were strongly dependent on DRI content. Basicity, which is the thermodynamic driving force of dephosphorization, decreased with the increasing DRI content because SiO{sub 2} concentration in the slag was proportional to DRI addition. The excess free energy of P{sub 2}O{sub 5} increased with the increasing SiO{sub 2} content in slag. A higher DRI content made dephosphorization difficult by decreasing the basicity and stability of P{sub 2}O{sub 5} in the slag. Therefore, when using DRI in EAF process, it is very important to control the basicity of slag.

36 MATERIALS SCIENCE↗
DOE OSTI2011

Lipidomic changes in rat liver after long-term exposure to ethanol

Alcoholic liver disease (ALD) is a serious health problem with significant morbidity and mortality. In this study we examined the progression of ALD along with lipidomic changes in rats fed ethanol for 2 and 3 months to understand the mechanism, and identify possible biomarkers. Male Fischer 344 rats were fed 5% ethanol or caloric equivalent of maltose-dextrin in a Lieber-DeCarli diet. Animals were killed at the end of 2 and 3 months and plasma and livers were collected. Portions of the liver were fixed for histological and immunohistological studies. Plasma and the liver lipids were extracted and analyzed by nuclear magnetic resonance (NMR) spectroscopy. A time dependent fatty infiltration was observed in the livers of ethanol-fed rats. Mild inflammation and oxidative stress were observed in some ethanol-fed rats at 3 months. The multivariate and principal component analysis of proton and phosphorus NMR spectroscopy data of extracted lipids from the plasma and livers showed segregation of ethanol-fed groups from the pair-fed controls. Significant hepatic lipids that were increased by ethanol exposure included fatty acids and triglycerides, whereas phosphatidylcholine (PC) decreased. However, both free fatty acids and PC decreased in the plasma. In liver lipids unsaturation of fatty acyl chains increased, contrary to plasma, where it decreased. Our studies confirm that over-accumulation of lipids in ethanol-induced liver steatosis accompanied by mild inflammation on long duration of ethanol exposure. Identified metabolic profile using NMR lipidomics could be further explored to establish biomarker signatures representing the etiopathogenesis, progression and/or severity of ALD. - Highlights: > Long term exposure to ethanol was studied. > A nuclear magnetic resonance (NMR) spectroscopy based lipidomic approach was used. > We examined the clustering pattern of the NMR data with principal component analysis. > NMR data were compared with histology and immunohistochemistry data. > Biochemical parameters were compared with the observed NMR lipid data.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2012

High rate mesophilic, thermophilic, and temperature phased anaerobic digestion of waste activated sludge: A pilot scale study

Highlights: Black-Right-Pointing-Pointer High temperatures were tested in single and two-stage anaerobic digestion of waste activated sludge. Black-Right-Pointing-Pointer The increased temperature demonstrated the possibility of improving typical yields of the conventional mesophilic process. Black-Right-Pointing-Pointer The temperature phased anaerobic digestion process (65 + 55 Degree-Sign C) showed the best performances with yields of 0.49 m{sup 3}/kgVS{sub fed}. Black-Right-Pointing-Pointer Ammonia and phosphate released from solids destruction determined the precipitation of struvite in the reactor. - Abstract: The paper reports the findings of a two-year pilot scale experimental trial for the mesophilic (35 Degree-Sign C), thermophilic (55 Degree-Sign C) and temperature phased (65 + 55 Degree-Sign C) anaerobic digestion of waste activated sludge. During the mesophilic and thermophilic runs, the reactor operated at an organic loading rate of 2.2 kgVS/m{sup 3}d and a hydraulic retention time of 20 days. In the temperature phased run, the first reactor operated at an organic loading rate of 15 kgVS/m{sup 3}d and a hydraulic retention time of 2 days while the second reactor operated at an organic loading rate of 2.2 kgVS/m{sup 3}d and a hydraulic retention time of 18 days (20 days for the whole temperature phased system). The performance of the reactor improved with increases in temperature. The COD removal increased from 35% in mesophilic conditions, to 45% in thermophilic conditions, and 55% in the two stage temperature phased system. As a consequence, the specific biogas production increased from 0.33 to 0.45 and to 0.49 m{sup 3}/kgVS{sub fed} at 35, 55, and 65 + 55 Degree-Sign C, respectively. The extreme thermophilic reactor working at 65 Degree-Sign C showed a high hydrolytic capability and a specific yield of 0.33 gCOD (soluble) per gVS{sub fed}. The effluent of the extreme thermophilic reactor showed an average concentration of soluble COD and volatile fatty acids of 20 and 9 g/l, respectively. Acetic and propionic acids were the main compounds found in the acids mixture. Because of the improved digestion efficiency, organic nitrogen and phosphorus were solubilised in the bulk. Their concentration, however, did not increase as expected because of the formation of salts of hydroxyapatite and struvite inside the reactor.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2010

Hydrothermal Detoxization of Slate Containing Asbestos and the Possibility of Application for Fertilizer of its Products

Hydrothermal decomposition of slate (building materials) containing asbestos has been attempted by using a NH{sub 4}H{sub 2}PO{sub 4} solution. Firstly, the alteration of chrysotile as a starting material was investigated under hydrothermal conditions of 200 deg. C, 12 hrs of reaction time and with a phosphate solution. It was confirmed that the original fibrous form of chrysotile had been perfectly collapsed by the SEM observation. The chrysotile (asbestos) disappeared to form Mg-Ca-Silicate (Ca{sub 7}Mg{sub 2}P{sub 6}O{sup 24}) estimated by XRD. The composition and chemical form of reaction products (Mg-Ca-Silicate) was predicted to application as a fertilizer. Fertilizer effect of these resulted product on cultivations of Japanese radish (leaves), soybeans and tomatoes, was examined by using a special medium of mixed soil with a low content of N, P, K and a thermal-treated zeolite one. The fertilizer effects of the product were compared to commercial fertilizers such as N, N-K-P and P types. In order to estimate the fertilizer effect, the size of crops, number of fruits and number of leaves were measured everyday. As a result, these hydrothermal products of slate containing asbestos were as good as commercial fertilizers on the market. Fruits groups especially had a good crop using the hydrothermal slate product. These results show that the main components of hydrothermal treatments slate are calcium silicate and magnesium phosphate. Its decomposition reaction products may have the possibility of application for fertilization of crops which require nucleic acid--phosphorus.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2016

Reaction paths of phosphine dissociation on silicon (001)

Using density functional theory and guided by extensive scanning tunneling microscopy (STM) image data, we formulate a detailed mechanism for the dissociation of phosphine (PH{sub 3}) molecules on the Si(001) surface at room temperature. We distinguish between a main sequence of dissociation that involves PH{sub 2}+H, PH+2H, and P+3H as observable intermediates, and a secondary sequence that gives rise to PH+H, P+2H, and isolated phosphorus adatoms. The latter sequence arises because PH{sub 2} fragments are surprisingly mobile on Si(001) and can diffuse away from the third hydrogen atom that makes up the PH{sub 3} stoichiometry. Our calculated activation energies describe the competition between diffusion and dissociation pathways and hence provide a comprehensive model for the numerous adsorbate species observed in STM experiments.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2021

Li interaction-induced phase transition from black to blue phosphorene

A comprehensive first-principle calculation has been carried out and revealed that at sufficiently high Li concentration and certain well-defined configurations a phase transition from black to blue phosphorene can take place. Blue phosphorene, a newly predicted allotrope of phosphorus, possesses unique crystalline and electronic structure and is a promising candidate, not only for fundamental research but also for electronic and optoelectronic applications. Methods to growth high quality blue phosphorene layers are highly desirable but challenging. Here, a novel kinetic pathway to grow blue phosphorene layers from black phosphorene layers via Li intercalation is proposed based on first principle study. This study pointed out that Li atoms intercalated in black phosphorene could act as ‘catalyst’ in the ‘reactive region’ of the lone pair of P atoms, leading to a P-P bond breaking and subsequently, a local structural transformation from orthorhombic lattice to an assembly of parallel narrow nanoribbons with rhombohedra-like symmetry. During Li deintercalation, these nanoribbons are self-mended and form blue phosphorene layers. The interlayer distance was found 4.60 Å for double layer with AA stacking and 4.13 Å for multilayer with ABC stacking, respectively, indicating a monolayer blue phosphorene can be mechanically exfoliated. Furthermore, this study also points out the possibility of new phases in other systems, where intercalation can lead to an unexpected structural phase transition and even a discovery of novel materials.

36 MATERIALS SCIENCE↗
DOE OSTI2018

The Fluorescent Properties and Electronic Structure of Nd(111) Complexes with Carboxylic Acids

Mixed-ligand neodymium(III) complexes with carboxylic acids and nitrogen- and phosphorus-containing neutral ligands, which luminesce in the near-IR region, are studied by luminescence and X-ray photoelectron spectroscopies. It is found that coordination of neutral ligands through the nitrogen donor atom leads to an increase in the electron density at the neodymium(III) ion.

74 ATOMIC AND MOLECULAR PHYSICS↗
DOE OSTI2015

Uranium Biomineralization by Natural Microbial Phosphatase Activities in the Subsurface

In this project, inter-disciplinary research activities were conducted in collaboration among investigators at The University of Alabama (UA), Georgia Institute of Technology (GT), Lawrence Berkeley National Laboratory (LBNL), Brookhaven National Laboratory (BNL), the DOE Joint Genome Institute (JGI), and the Stanford Synchrotron Radiation Light source (SSRL) to: (i) confirm that phosphatase activities of subsurface bacteria in Area 2 and 3 from the Oak Ridge Field Research Center result in solid U-phosphate precipitation in aerobic and anaerobic conditions; (ii) investigate the eventual competition between uranium biomineralization via U-phosphate precipitation and uranium bioreduction; (iii) determine subsurface microbial community structure changes of Area 2 soils following organophosphate amendments; (iv) obtain the complete genome sequences of the Rahnella sp. Y9-602 and the type-strain Rahnella aquatilis ATCC 33071 isolated from these soils; (v) determine if polyphosphate accumulation and phytate hydrolysis can be used to promote U(VI) biomineralization in subsurface sediments; (vi) characterize the effect of uranium on phytate hydrolysis by a new microorganism isolated from uranium-contaminated sediments; (vii) utilize positron-emission tomography to label and track metabolically-active bacteria in soil columns, and (viii) study the stability of the uranium phosphate mineral product. Microarray analyses and mineral precipitation characterizations were conducted in collaboration with DOE SBR-funded investigators at LBNL. Thus, microbial phosphorus metabolism has been shown to have a contributing role to uranium immobilization in the subsurface.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2018

Leaf Area, Vegetation Biomass and Nutrient Content, Utqiagvik (Barrow), Alaska, 2012 - 2013

This dataset consists of measurements of vegetation harvested from Areas A to D of Intensive Site 1 at the Next-Generation Ecosystem Experiments (NGEE) Arctic site near Barrow, Alaska. The dataset includes i) values of leaf area index, biomass, carbon (C), nitrogen (N) and phosphorus (P) content of aboveground plant parts from 0.25 m x 0.25 m clip-plots at peak growing season and ii) fine-root biomass from 5.08-cm diameter soil cores taken throughout the active layer in the same location as the clip plots in late July-early August 2012, and iii) values of aboveground biomass and nitrogen (N) content measured from 0.1 m x 0.1 m clip-plots harvested at 2-week intervals throughout the 2013 growing season. This dataset includes five csv files and one pdf file.The Next-Generation Ecosystem Experiments: Arctic (NGEE Arctic), was a research effort to reduce uncertainty in Earth System Models by developing a predictive understanding of carbon-rich Arctic ecosystems and feedbacks to climate. NGEE Arctic was supported by the Department of Energy's Office of Biological and Environmental Research.The NGEE Arctic project had two field research sites: 1) located within the Arctic polygonal tundra coastal region on the Barrow Environmental Observatory (BEO) and the North Slope near Utqiagvik (Barrow), Alaska and 2) multiple areas on the discontinuous permafrost region of the Seward Peninsula north of Nome, Alaska.Through observations, experiments, and synthesis with existing datasets, NGEE Arctic provided an enhanced knowledge base for multi-scale modeling and contributed to improved process representation at global pan-Arctic scales within the Department of Energy's Earth system Model (the Energy Exascale Earth System Model, or E3SM), and specifically within the E3SM Land Model component (ELM).

%_nitrogen↗
DOE OSTI2011

A general solution-chemistry route to the synthesis LiMPO{sub 4} (M=Mn, Fe, and Co) nanocrystals with [010] orientation for lithium ion batteries

A general and efficient solvothermal strategy has been developed for the preparation of lithium transition metal phosphate microstructures (LiMnPO{sub 4}, LiFePO{sub 4}, and LiCoPO{sub 4}), employing ethanol as the solvent, LiI as the Li source, metal salts as the M sources, H{sub 3}PO{sub 4} as the phosphorus source, and poly(vinyl pyrrolidone) (PVP) as the carbon source and template. This route features low cost, environmental benign, and one-step process for the cathode material production of Li-ion batteries without any complicated experimental setups and sophisticated operations. The as-synthesized LiMPO{sub 4} microstructures exhibit unique, well-shaped and favorable structures, which are self-assembled from microplates or microrods. The b axis is the preferred crystal growth orientation of the products, resulting in a shorter lithium ion diffusion path. The LiFePO{sub 4} microstructures show an excellent cycling stability without capacity fading up to 50 cycles when they are used as a cathode material in lithium-ion batteries. - Graphical abstract: A general and efficient solvothermal strategy has been developed for the preparation of lithium transition metal phosphate microstructures under solvothermal conditions in the presence of PVP. Highlights: > A general and efficient solvothermal strategy has been developed for the preparation of LiMPO{sub 4} microstructures. > This route features low cost, environmental benign, and one-step process. > The LiMPO{sub 4} microstructures exhibit unique, well-shaped, and favorable structures. > The LiFePO{sub 4} microstructures show an excellent cycling stability up to 50 cycles as a cathode material of lithium-ion batteries.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2023

Experimental and theoretical investigations of rotating algae biofilm reactors (RABRs): Areal productivity, nutrient recovery, and energy efficiency

Microalgae biofilms have been demonstrated to recover nutrients from wastewater and serve as biomass feedstock for bioproducts. However, there is a need to develop a platform to quantitatively describe microalgae biofilm production, which can provide guidance and insights for improving biomass areal productivity and nutrient uptake efficiency. Here, this paper proposes a unified experimental and theoretical framework to investigate algae biofilm growth on a rotating algae biofilm reactor (RABR). Experimental laboratory setups are used to conduct controlled experiments on testing environmental and operational factors for RABRs. We propose a differential–integral equation‐based mathematical model for microalgae biofilm cultivation guided by laboratory experimental findings. The predictive mathematical model development is coordinated with laboratory experiments of biofilm areal productivity associated with ammonia and inorganic phosphorus uptake by RABRs. The unified experimental and theoretical tool is used to investigate the effects of RABR rotating velocity, duty cycle (DC), and light intensity on algae biofilm growth, areal productivity, nutrient uptake efficiency, and energy efficiency in wastewater treatment. Our framework indicates that maintaining a reasonable light intensity range improves biomass areal productivity and nutrient uptake efficiency. Our framework also indicates that faster RABR rotation benefits biomass areal productivity. However, maximizing the nutrient uptake efficiency requires a reasonably low RABR rotating speed. Energy efficiency is strongly correlated with RABR rotating speed and DC.

09 BIOMASS FUELS↗
DOE OSTI2022

Stronger fertilization effects on aboveground versus belowground plant properties across nine U.S. grasslands

Abstract Increased nutrient inputs due to anthropogenic activity are expected to increase primary productivity across terrestrial ecosystems, but changes in allocation aboveground versus belowground with nutrient addition have different implications for soil carbon (C) storage. Thus, given that roots are major contributors to soil C storage, understanding belowground net primary productivity (BNPP) and biomass responses to changes in nutrient availability is essential to predicting carbon–climate feedbacks in the context of interacting global environmental changes. To address this knowledge gap, we tested whether a decade of nitrogen (N) and phosphorus (P) fertilization consistently influenced aboveground and belowground biomass and productivity at nine grassland sites spanning a wide range of climatic and edaphic conditions in the continental United States. Fertilization effects were strong aboveground, with both N and P addition stimulating aboveground biomass at nearly all sites (by 30% and 36%, respectively, on average). P addition consistently increased root production (by 15% on average), whereas other belowground responses to fertilization were more variable, ranging from positive to negative across sites. Site‐specific responses to P were not predicted by the measured covariates. Atmospheric N deposition mediated the effect of N fertilization on root biomass and turnover. Specifically, atmospheric N deposition was positively correlated with root turnover rates, and this relationship was amplified with N addition. Nitrogen addition increased root biomass at sites with low N deposition but decreased it at sites with high N deposition. Overall, these results suggest that the effects of nutrient supply on belowground plant properties are context dependent, particularly with regard to background N supply rates, demonstrating that site conditions must be considered when predicting how grassland ecosystems will respond to increased nutrient loading from anthropogenic activity.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

High-yield recombinant bacterial expression of 13 C-, 15 N-labeled, serine-16 phosphorylated, murine amelogenin using a modified third generation genetic code expansion protocol

Amelogenin constitutes ~ 90% of the enamel matrix in the secretory stage of amelogenesis, a still poorly understood process that results in the formation of the hardest and most mineralized tissue in vertebrates – enamel. Most biophysical research with amelogenin uses recombinant protein expressed in Escherichia coli. In addition to providing copious amounts of protein, recombinant expression allows 13 C- and 15 N-labelling for detailed structural studies using NMR spectroscopy. However, native amelogenin is phosphorylated at one position, Ser-16 in murine amelogenin, and a lingering question overshadowing decades of enamel research with recombinant amelogenin is the effect of phosphorylation on its biophysical properties. Using a modified genetic code expansion protocol we have expressed and purified uniformly 13 C-, 15 N-labeled murine amelogenin (pS16M179) with ~95% of the protein being correctly phosphorylated. Homogeneous phosphorylation was achieved using commercially available, enriched, 13 C-, 15 N-labeled media and protein expression was induced with isopropyl β-D-1-thiogalactopyranoside at 310 K. Phosphoserine incorporation was verified from one-dimensional 31P NMR spectra, comparison of 1 H- 15 N HSQC spectra, Phos-tag SDS PAGE, and mass spectrometry. Phosphorus-31 NMR spectra for pS16M179 under conditions known to trigger amelogenin self-assembly into nanospheres confirm nanosphere models with buried N-termini. Lambda phosphatase treatment of these nanospheres results in the dephosphorylation of pS16M179, confirming that smaller oligomers and monomers with exposed N-termini are in equilibrium with nanospheres. Such 13 C-, 15 N-labeling of amelogenin with accurately encoded phosphoserine incorporation will accelerate biomineralization research to understand amelogenesis and stimulate the expanded use of genetic code expansion protocols to introduce phosphorylated amino acids into proteins.

31P-NMR↗
DOE OSTI2019

Long-Term Effects of Crude Oil Spillage on Selected Physicochemical Properties Including Heavy Metal Contents of Sandy Tropical Soil

Crude oil spillage effects on the environment often wane with time, making late remediation of affected soils look irrelevant. Physicochemical quality of a sandy soil under 9-year-old spillage was compared with that of adjacent unaffected site in southern Nigeria. Soil bulk density and equilibrated water content were higher in affected than unaffected site, but permeability did not change. The spillage increased soil pH, organic carbon, total nitrogen and available phosphorus by about 7%, 1700%, 133% and − 16%, respectively. It lowered divalent exchangeable bases/acidity but raised base saturation. It increased total petroleum hydrocarbon (PHC{sub t}) and micronutrients/heavy metals (Fe, Mn, Zn, Cd and Pb), all of which were below their critical limits in soils by regulatory bodies. Soil pH, organic carbon and PHC{sub t} correlated positively with all five micronutrients/heavy metals; total nitrogen did so with Zn and Pb. Nine-year period may be insufficient for spillage effects in sandy soils to cease to be evident. Such effects for PHC{sub t} and heavy metals, however, are deemed tolerable for ecological safety.

54 ENVIRONMENTAL SCIENCES↗