Ammonium Sulfate Concentration Research Articles

Time-course analysis of the transcriptome of Arabidopsis thaliana leaves under high-concentration ammonium sulfate treatment

ABSTRACT Ammonium fertilization is of great interest for future agriculture, as unlike nitrate, ammonium use efficiency does not decrease in C3 plants, even in environments with elevated CO2 concentrations. However, excess ammonium often results in toxic effects such as growth suppression and chlorosis, i.e. ammonium toxicity. The addition of nitrate, a major source of nitrogen commonly found in soils, has been shown to alleviate these toxic effects. Understanding the mechanisms of ammonium toxicity in the presence of nitrate is crucial for the effective use of ammonium fertilizers in crop cultivation. To elucidate these responses, a time-course analysis of the transcriptome was performed on A. thaliana leaves treated with high concentrations of ammonium sulfate in the presence of sufficient nitrate. The expression of nitrate-inducible genes tended to be downregulated by the treatment. The expression of genes relating to abscisic acid (ABA), jasmonic acid (JA), salicylic acid (SA), and membrane trafficking was upregulated, whereas that of photosynthesis-, auxin-, and cytokinin-related genes involved in growth and development was downregulated. The induction of many osmotic stress-responsive genes and nitric oxide (NO)-inducible genes suggests the involvement of osmotic stress and NO signaling in the response. This study provides a novel and comprehensive overview of transcriptional changes occurring in response to high ammonium sulfate concentrations with sufficient nitrate, and sheds light on potential pathways involved in ammonium toxicity under these conditions.

Optimizing pectin lyase production using the one-factor-at-a-time method and response surface methodology.

Pectinases are commonly industrially synthesized by molds. This study aimed to optimize pectin lyase synthesis by a bacterium, Pseudomonas fluorescens, using both the one-factor-at-a-time (OFAT) method and response surface methodology. First, on optimization of pectin lyase fermentation by the OFAT method, the effects of pectin, peptone, yeast extract, (NH4)2SO4, pH, and salts were investigated. The highest pectin lyase activity was determined to be 28.63U/mL at pH 8, 30°C, with 1% (w/v) pectin and 0.14% (w/v) (NH4)2SO4 concentration at the 90th hour. The effect of substrate inhibition on the microbial growth was also investigated, and the results showed that the process can be described by noncompetitive inhibition model. The values of kinetic constants were determined as µm=0.175h-1, KS=6.931g/L, and, KI=6.932g/L by nonlinear regression analysis. It was reported that pectin lyase enzymes exhibited peak activity at 50°C and pH 8. Finally, response surface methodology (RSM) was utilized to optimize pH, concentrations of ammonium sulfate, and pectin, which were chosen as independent variables. The interactions between these variables were also examined. According to RSM, the optimum values of the parameters to achieve a maximum pectin lyase activity of 35.62U/mL were determined to be pH 7.97, 1.25% (w/v) pectin concentration, and 0.25% (w/v) (NH4)2SO4 concentration.

Partial biochemical and immunological characterization of an isolated 52.1 kDa pregnancy-associated glycoprotein charge variant

Pregnancy-associated glycoproteins (PAGs) are synthesized in the placental cells of ruminants and are detectable in blood, milk, and urine. Many of these proteins have been obtained and characterized from placental extracts by precipitation with 80 % ammonium sulfate. The possibility of purifying PAGs by precipitation with other concentrations of ammonium sulfate remains unexplored. We aimed to study PAG proteins obtained from extracts of ovine placenta at 100 days of gestation through precipitation with 40 % ammonium sulfate (Extract 40). The main protein complex (130 kDa) was obtained after Extract 40 precipitation. Under reducing SDS-PAGE conditions, the 130 kDa complex dissociated in two PAG proteins with apparent molecular weights of 52.1 kDa and 26.1 kDa. The 130 kDa protein appeared to be a molecular complex consisting of two copies of the 52.1 kDa protein linked to one copy of the 26.1 kDa protein, presumably by disulfide bonds. Furthermore, the 52.1 kDa protein consisted of at least three isoforms with distinct isoelectric points. Amino acid microsequencing of the 52.1 kDa protein revealed a chimeric structure containing amino acid sequences of PAG1, PAG4, PAG6, and PAG1-like proteins. This procedure recovered a novel 130 kDa protein complex composed of 26.1 kDa and two 52.1 kDa PAGs. To the best of our knowledge, this has not been previously reported as heterologous polymeric molecules.

Leaching Thermodynamics of Low-Grade Copper Oxide Ore from [(NH4)2SO4]-NH3-H2O Solution.

This paper describes a highly alkaline low-grade copper oxide ore. Copper can be selectively leached out while other metals are retained. A thermodynamic model of the CuO-(NH4)2SO4-NH3-H2O system was established for the leaching of tenorite (CuO) under conditions of mass and charge conservation. MATLAB's fitting functions, along with the diff and solve functions, were used to calculate the optimal ammonia concentration and total copper ion concentration of tenorite under different ammonium sulfate concentrations. The effects of various ammonia-ammonium salt solutions (ammonium sulfate, ammonium carbonate, ammonium chloride) on the copper leaching rate were investigated. Results show that under the conditions of an ammonia concentration of 1.2 mol/L, an ammonia-ammonium ratio of 2:1, a liquid-solid ratio of 3:1, a temperature of 25 °C, and a leaching time of 4 h, the copper leaching rate from the ammonium sulfate and ammonium chloride solutions reaches 70%, which is slightly higher than that of ammonium carbonate. Therefore, an ammonia-ammonium sulfate system is selected for leaching low-grade copper oxide due to its lower corrosion to equipment compared to the chlorination system. The impact of this study on industrial applications includes the potential to find more sustainable and cost-effective methods for resource recovery. The industry can reduce its dependence on resources and mitigate its environmental impact. Readers engaged in low-grade oxidized copper research will benefit from this study.

Towards circular economy of wasted printed circuit boards of mobile phones fuelled by machine learning and robust mathematical optimization framework

Estimating the operating conditions using conventional process analysis techniques for the maximum metal extraction from the wasted printed circuit boards (WPCB) can provide sub-optimal solutions leading to the low yield of the process. In this paper, we present a closed-loop methodological framework built on machine learning and robust mathematical optimization technique, that offers the mathematical rigour, to determine the optimum operating conditions for the maximum Cu and Ni recovery from the WPCB. Alkali leaching based novel metals recovery process from the WPCB is designed, and the experiments are conducted to collect the data on the percentage recovery of Cu and Ni against the operating levels of the process input variables (ammonia concentration (NH3 conc. (g/L)), ammonium sulfate concentration ((NH4)2SO4 conc. (g/L)), H2O2 concentration (H2O2 conc. (M)), time (h), liquid to solid ratio (L/S ratio, (mL/g)), temperature (Temp. (°C)), and stirring speed (rpm)). The experimental data is deployed to construct the functional mapping between the nonlinear output variables of metals recovery process with the hyperdimensional input space through artificial neural network (ANN) based modelling algorithm – a powerful universal function approximator. Well-predictive ANN models for Cu and Ni recovery are developed having co-efficient of determination (R2) value more than 0.90. Partial derivative-based sensitivity analysis is then carried out to establish the order of the significance of the input variables that is backed by the domain knowledge, thus promotes the interpretability of the trained ANN models. The hybridization of ANN with NLP (nonlinear programming) framework is implemented for the determination of optimized operating conditions to extract maximum Cu and Ni under separate and combined model of metal extraction. The robustness of the determined solutions is verified, the determined optimized solutions for the metal recovery are validated in the lab, and the maximum metal recovery, i.e., 100 % Cu and 90 % Ni is extracted from the WPCB. This research demonstrates the effective utilization of ANN model-based robust optimization approach for the metal recovery from the WPCB that supports the circular economy for the metal extraction industry.

Study on the role of microbial metabolites in in-situ noncontact bioleaching of ion-adsorption rare earth ore

Ion adsorption rare earth ore nearly satisfy global market demand for heavy rare earth elements (HREEs). Bio-leaching has important potential for the clean and efficient extraction of ion-adsorption rare earth ore. However, the complexities of in-situ mining restrict the use of contact/direct bio-leaching, and non-contact/indirect bio-leaching would be the best choice. This study explore the potential of fermentation broths prepared by Yarrowia lipolytica (ATCC 30162) for the bio-leaching of ion-adsorption rare earth ore, and three typical metabolites (potassium citrate (K3Cit), sodium citrate (Na3Cit) and ammonium citrate ((NH4)3Cit) of Yarrowia lipolytica were further evaluated in simulated bioleaching (non-contact bioleaching) of ion-adsorption rare earth ore, including leaching behavior, seepage rule and rare earth elements (REEs) morphological transformation. The column leaching experiments shown that direct leaching of REEs using fermentation broths results in incomplete leaching of REEs due to the influence of impurities. Using the purified and prepared metabolites as lixiviant, REEs can be effectively extracted (leaching efficiency >90%) at cation concentration was only 10 % of the commonly used ammonium sulfate concentration (45 mM). Cation type had less effect on leaching efficiency. During the ion-adsorption rare earth ore leaching process, rare earth ions form a variety of complex chelates with citrate, thus transferring rare earth elements from the mineral surface to the leachate. Experimental results showed that pH and concentration together determined the type and form of rare earth chelates, which in turn affect the leaching behavior of REEs and solution seepage rule. This study helps to provide a theoretical basis for the regulation and enhancement of ion-adsorption rare earth ore non-contact bioleaching process.

Real-time dynamic measurement of scattering characteristics of optically-levitated fly ash interacting with water-soluble ammonium sulfate aerosol

This paper proposes a coupled laser levitation and scattering measuring system to realize real-time dynamic measurement of scattering characteristics of individual fly ash particles interacting with water-soluble ammonium sulfate aerosols of different concentrations. The scattering coefficients, absorption coefficients, and phase functions of aerosols were calculated. The disparities in scattering characteristics interacting fly ash with ammonium sulfate, and the impact of typical influencing parameters on the scattering characteristics were discussed in detail. The findings highlighted that the scattering characteristics of aerosols followed with those of Mie particles, with an increase in the particle size leading to an enhancement in the forward scattering intensity. Notably, dynamic measurement demonstrated that the scattering intensity with high ammonium sulfate concentrations (≥1 mol/L) showed an upward trend, exerting a more pronounced impact than low-concentration counterparts. Furthermore, the particle size has a stronger effect on the scattering characteristics than the complex refractive index induced by varying concentrations.

Dispersion polymerization of AM-AMPS-DAM-DOH and its sand control property

Sand production in oil wells is a common challenge during the oil extraction process. Water-soluble polymers have attracted attention due to their low toxicity, cost-effectiveness, ease of direct injection, and excellent thermal stability. However, the dry polymer injection process commonly used in offshore oil fields is complex. Therefore, this paper employs acrylamide (AM), 2-acrylamide-2-methylpropane sulfonic acid (AMPS), a dihydroxy monomer (DOH), and a functional monomer (DAM) containing ketone groups and self-crosslinking properties as monomers. A polymer latex type sand control agent was prepared through dispersion polymerization for direct application on offshore platforms. This paper utilizes flocculation time and maximum sand free flow rate as inspection indicators and utilizes both the one-factor-at-a-time method and the surface response method to optimize the synthesis conditions of the sand control agent. The optimal synthesis conditions are obtained: total monomer concentration of 9.15 wt%, DOH concentration at 10 wt% of the total monomer concentration, ammonium sulfate (AS) concentration of 27.02 wt%, dispersant concentration of 2.13 wt%, initiator concentration at 0.25 wt% of the total monomer concentration, reaction time of 12 h, and reaction temperature of 40 °C. Under the conditions of concentration ≥2000 mg/L and curing time ≥4 h, the sand control agent is suitable for sand stabilization under formation conditions of temperatures between 50 and 80 °C, pH levels ranging from 3 to 9, salinity <30000 mg/L, and sand particle size of ≥100 μm. The research results of this paper can serve as a valuable reference for the control of oilfield sand production.

Production, purification and characterization of endo-polygalacturonase using novel strain of Bacillus pumilus through RSM

Polygalacturonases (PG) are the enzymes that cause depolymerization of pectin. In current study, bacterial strain was isolated from rotten sample and then purified. It was screened for endo-polygalacturonase activity using PSAM media. It had shown positive response for endo-PG activity. Bacterial DNA was isolated and 16 S rRNA gene amplification was done using universal primer pair of 8 F and 1492 R. Bacterial strain was also amplified by 16 S rRNA gene for sequencing. BLAST of gene sequence on NCBI database had shown that bacterial isolate was identified as Bacillus pumilus. Endo-polygalacturonase enzyme was produced by submerged fermentation to optimize culture conditions by RSM in JMP-12 software. The optimized parameters had shown maximum endo-polygalacturonase activity of 153.2 U/mL/min after using 3 g of orange peels as substrate, 3 mL of inoculum size, 6.5 pH buffer, 40°C temperature and 3 days of incubation. After optimizing fermentation conditions, endo-polygalacturonase was precipitated using 70 % concentration of ammonium sulfate. This partially precipitated sample was dialyzed with dialysis tube and used to find activity of endo-polygalacturonase (1620.6 U/mL/min). This sample was applied to gel filtration chromatography for further purification. Endo-polygalacturonase activity increased many times 2231.44 U/mL/min after gel filtration. The Molecular weight of endo-polygalacturonase was 46 kDa after SDS PAGE characterization. High Vmax value and alkaliphilic nature of endo-polygalacturonase will make it good candidate for food and feed industry near future.

Efficient experimental method for purifying allergens from aqueous extracts

Studying the molecular and immunological basis of allergic diseases often requires purified native allergens. The methodologies for protein purification are usually difficult and may not be completely successful. The objective of this work was to describe a methodology to purify allergens from their natural source, while maintaining their native form. The purification strategy consists of a three-step protocol and was used for purifying five specific allergens, Ole e 1, Amb a 1, Alt a 1, Bet v 1 and Cup a 1.Total proteins were extracted in PBS (pH 7.2). Then, the target allergens were pre-purified and enriched by salting-out using increasing concentrations of ammonium sulfate. The allergens were further purified by anion exchange chromatography. Purification of Amb a 1 required an extra step of cation exchange chromatography. The detection of the allergens in the fractions obtained were screened by SDS-PAGE, and Western blot when needed. Further characterization of purified Amb a 1 was performed by mass spectrometry. Ole e 1, Alt a 1, Bet v 1 and Cup a 1 were obtained at > 90 % purity. Amb a 1 was obtained at > 85 % purity.Overall, we propose an easy-to-perform purification approach that allows obtaining highly pure allergens. Since it does not involve neither chaotropic nor organic reagents, we anticipate that the structural and biological functions of the purified molecule remain intact. This method provides a basis for native allergen purification that can be tailored according to specific needs.

Conversion of Sweet Whey to Bioethanol: A Bioremediation Alternative for Dairy Industry

In many countries, whey from the dairy industry is an abundant waste that generates an important environmental impact. Alternative processes to use the whey and minimize the environmental impact are needed. This work considered six formulations with different ammonium sulfate and L-phenylalanine (L-Phe) concentrations to produce bioethanol in sweet whey fermentation by Kluyveromyces marxianus. The results showed a maximum bioethanol concentration equal to 25.13 ± 0.37 g L−1 (p &lt; 0.05) for formulation F6, with 1 g L−1 of L-Phe and 1.350 g L−1 of ammonium sulfate (96 h). For these conditions, the chemical oxygen demand removal percentage (CODR%) was 67%. The maximum CODR% obtained was 97.5% for formulation F3 (1 g L−1 of L-Phe) at 96 h; however, a significant decrease in bioethanol concentration (14.33 ± 2.58 g L−1) was observed. On the other hand, for formulation, F3, at 48 h of fermentation time, a bioethanol concentration of 23.71 ± 1.26 g L−1 was observed, with 76.5% CODR%. Based on these results, we suggest that the best conditions to obtain a significant bioethanol concentration and CODR% value are those used on the configuration F3 at 48 h.

Cellulases production optimization and inductors evaluation in pretreated plantain rachis by Penicillium oxalicum

AbstractPretreated lignocellulosic residues are suitable substrates for cellulases production by filamentous fungi. In the current work, plantain rachis was pretreated with sequential acid and alkali and then used as the main carbon source for cellulases production. First, a full 23 factorial design and response surface methodology (RSM), based on central composite rotatable design (CCRD), were carried out to cellulases production media optimization from plantain rachis by Penicillium oxalicum. The cellulases production was evaluated in flasks and bioreactor scale; in parallel, the addition of possible cellulases inductors was evaluated in flasks: molasses, beer bran, oat bran, and wheat bran. Results from statistical analyses with a level of confidence of 95% demonstrated that the concentration of ammonium sulfate must be kept at 1.625 g/L. The optimum urea and yeast extract concentrations were 0.560 g/L and 0.250 g/L, respectively. Cellulases volumetric productivities were higher in instrumented bioreactor than in flasks: 78.03% for exoglucanase, 10.87% for endoglucanase, 1.58% for β‐glucosidase, and 44.36% for FPU. Therefore, P. oxalicum was able to produce cellulases from plantain rachis in flasks and bioreactor, and molasses was the additional inductor that presented an increment in cellulases activities: endoglucanase 15%, exoglucanase 81%, and β‐glucosidase 55%.

Nitrification inhibitors impose distinct effects on comammox bacteria and canonical ammonia oxidizers under high N fertilization regimes

The response of complete ammonia-oxidizing (comammox) bacteria to variable nitrogen (N) inputs and nitrification inhibitors (NIs) remains poorly understood. We conducted two experiments, in a soil where both clade A and B comammox Nitrospira were present, to identify the type (ammonium sulphate vs. urea) and the level of N-fertilization (0–500 mg-N-kg‐1d.w. soil) that are conducive to the growth of comammox, compared to canonical ammonia-oxidizing microorganisms (AOM), and subsequently to assess their response to three commercial (DCD, nitrapyrin, DMPP) and a new potent (ethoxyquin) NI under these favourable N fertilization regimes. The highest concentration of ammonium sulphate stimulated comammox compared to canonical AOM which showed a dose-dependent increase by both N sources. DMPP primarily, and DCD, imposed a more persistent inhibition on nitrification than nitrapyrin and ethoxyquin, a result which did not concur with the soil persistence of NIs (DMPP was the least persistent among the commercial NIs) but was mostly driven by their selective inhibitory activity on ammonia-oxidizing bacteria (AOB) that prevailed in the studied soil. The inhibition of AOB by DMPP and DCD was complemented by the proliferation of comammox and ammonia-oxidizing archaea (AOA). NIs induced significant changes on AOB and comammox communities. Comammox Nitrospira clade B were enriched by DCD and DMPP but depleted by nitrapyrin and ethoxyquin. We showed that (i) releasing comammox Nitrospira and AOA from AOB competition allows them to proliferate efficiently, even under high inorganic ammonium conditions; (ii) comammox clade B are more responsive than clade A to high inorganic N input and NIs.

New experimental findings on the separation of cobalt and nickel from an ammonia-ammonium-based leach liquor using ammonium-saponified Cyanex 272

There is a need to improve cobalt and nickel recovery from industrial waste streams such as secondary leach solutions and rechargeable battery leach solutions due to the increasing economic value of these critical metals. Ammonia is an important lixiviant due to its limited environmental impact compared to acid based lixiviants, however, the subsequent selective extraction of cobalt and nickel from the leach liquor is a major challenge. This study presents new experimental findings on the treatment of a Base metal Refinery (BMR) secondary leach liquor that contains high concentrations of ammonia and ammonium sulphate, and low concentrations of cobalt (∼442 mg/L) and nickel (∼1624 mg/L) using partially saponified Cyanex 272. New knowledge that can be applied in the treatment of ammonia based leachates from various industrial wastes that contain residual cobalt and nickel is presented herein. The investigation was conducted through the determination of the optimum extraction temperature, percentage saponification, and organic to aqueous ratios. Partial saponification of Cyanex 272 increased ammonia-metal complex stability at the interphase and this limited cobalt extraction between 30 °C and 50 °C. However, a high cobalt extraction percentage, and a high separation factor of 94.07 %, and 1189.76 respectively were achieved at 60 °C, 20 % saponification, and 0.5 O/A loadings in a single contact. A second-stage batch extraction produced a nickel aqueous stream with no cobalt. Stripping of the loaded organic phase with dilute sulphuric acid produced a pure cobalt aqueous stream. Further, the extractant was successfully regenerated post stripping. Overall, partial ammonium-saponification increased the extent of cobalt extraction, and selectivity. The novel process conditions reported herein can be used to design sustainable cobalt-nickel separation processes.

Ultrasound Assisted Aqueous Two‐Phase Extraction of Polysaccharides from Ophiopogon japonicus: Process Optimization, Structure Characterization, and Antioxidant Activity

AbstractUltrasound assisted aqueous two‐phase extraction (UATPE) is used to extract polysaccharides from Ophiopogon japonicus (OJPs). Experimental factors in UATPE process are optimized by response surface methodology. The optimal extraction conditions are obtained as follows: the ethanol concentration of 34%, ammonium sulfate concentration of 23%, solid‐to‐liquid ratio of 1:17 g/mL, and ultrasound power of 480 W, and the yield of OJPs is 9.41% ± 0.12%, and then the crude OJPs are further purified by different column chromatography to obtain a homogenous fraction (OJPs‐2‐SG), and identified its structure via different methods. OJPs‐2‐SG with a molecular weight of 1.25 × 105 Da is comprised of mannose (Man), galactose (Gal), glucose (Glc), and rhamnose (Rha) with a molar ratio of 5.15: 26.39: 68.72:8.41. Moreover, OJPs‐2‐SG contained α and β glycosidic bonds, and didn't showed a triple helix structure. Furthermore, OJPs‐2‐SG described irregular structures, and cylindrical and conical block like features. The IC50 values of scavenging 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH), hydroxyl (OH), and superoxide (O2‐) radicals for OJPs‐2‐SG achieved 3.72 mg/mL, 3.89 mg/mL, and 8.71 mg/mL, respectively. These results provide important references for the extraction and application of polysaccharides.

Optimization of a Rare Earth and Aluminum Leaching Process from Weathered Crust Elution-Deposited Rare Earth Ore with Surfactant CTAB

Ammonium sulfate is typically employed as a leaching agent in the in situ leaching of weathered crust elution-deposited rare earth ore. However, it is associated with challenges such as low efficiency in mass transfer for rare earth (RE) leaching, high usage of the leaching agent, and prolonged leaching duration. To address the issues mentioned above, the surfactant cetyltrimethyl ammonium bromide (CTAB) was compounded with 2% ammonium sulfate to form a leaching agent in this paper. The effects of CTAB concentration, temperature, pH, and leaching agent flow rate on the rare earth (RE) and aluminum (Al) leaching mass transfer process from RE ore were investigated using chromatographic plate theory. The results revealed that CTAB addition improved the RE mass transfer process while moderately inhibiting the Al mass transfer efficiency. Increasing the temperature and pH of the leaching solution led to higher theoretical plate numbers for RE and Al leaching, lowered theoretical plate height (HETP), and enhanced leaching mass transfer efficiency. However, under high temperature and alkaline conditions, the mass transfer efficiency begins to decrease, indicating that high temperature and alkaline conditions are not conducive to the synergistic enhancement of RE and Al leaching by CTAB. Considering that clay minerals have good pH buffering properties, adjusting the pH of the leaching solution during rare earth ore leaching operations was deemed unnecessary. The optimal mass transfer conditions for leaching RE and Al were identified as 2% ammonium sulfate concentration, 0.00103 mol/L CTAB concentration, pH range of 5.2–5.5 for the leaching solution, 0.6 mL/min leaching solution flow rate, and room temperature. The rare earth leaching mass transfer effect could be enhanced during summer operations.

Prediction of Milk Powder Raman Spectra and Ammonium Sulfate Adulteration Concentration Based on Neural Network Model

This article aims to explore the changes in Raman spectra of milk powder when different concentrations of ammonium sulfate are adulterated. By conducting neural network tests on milk powder samples with different ammonium sulfate concentrations, we studied the impact of ammonium sulfate adulteration on milk powder. The research results show that moderate concentrations of ammonium sulfate adulteration can improve the solubility and flowability of milk powder, but high concentrations of adulteration may reduce the protein content of milk powder and have negative effects on its sensory quality and safety, causing significant adverse effects on human health. Therefore, the adulteration concentration of ammonium sulfate should be strictly controlled in the production and processing of milk powder to ensure its quality and safety.

PH-dependent reaction triggering in PmHMGR crystals for time-resolved crystallography

Serial crystallography and time-resolved data collection can readily be employed to investigate the catalytic mechanism of Pseudomonas mevalonii 3-hydroxy-3-methylglutaryl (HMG)-coenzyme-A (CoA) reductase (PmHMGR) by changing the environmental conditions in the crystal and so manipulating the reaction rate. This enzyme uses a complex mechanism to convert mevalonate to HMG-CoA using the co-substrate CoA and cofactor NAD+. The multi-step reaction mechanism involves an exchange of bound NAD+ and large conformational changes by a 50-residue subdomain. The enzymatic reaction can be run in both forward and reverse directions in solution and is catalytically active in the crystal for multiple reaction steps. Initially, the enzyme was found to be inactive in the crystal starting with bound mevalonate, CoA, and NAD+. To observe the reaction from this direction, we examined the effects of crystallization buffer constituents and pH on enzyme turnover, discovering a strong inhibition in the crystallization buffer and a controllable increase in enzyme turnover as a function of pH. The inhibition is dependent on ionic concentration of the crystallization precipitant ammonium sulfate but independent of its ionic composition. Crystallographic studies show that the observed inhibition only affects the oxidation of mevalonate but not the subsequent reactions of the intermediate mevaldehyde. Calculations of the pKa values for the enzyme active site residues suggest that the effect of pH on turnover is due to the changing protonation state of His381. We have now exploited the changes in ionic inhibition in combination with the pH-dependent increase in turnover as a novel approach for triggering the PmHMGR reaction in crystals and capturing information about its intermediate states along the reaction pathway.

Ultrasound-assisted Aqueous Two-Phase Extraction of Flavonoids from Scutellariae Radix and Evaluation of their Bioactivities In Vitro

Background: Scutellariae Radix, one of the most widely used herbs in Traditional Chinese Medicine, exhibits various biological activities due to its chemical components, which stand out for a number of flavonoids. In this study, Ultrasound-assisted aqueous two-phase extraction (UAATPE) was employed for the first time to obtain a high extraction rate and high purity of flavonoids from Scutellariae Radix. Methods: The Box-Behnken response surface method (RSM) was utilized to optimize the extraction conditions with the application of the new aqueous two-phase system (ATPS) composed of ethanol and ammonium sulfate. The major influence factors, including ethanol concentration, ammonium sulfate concentration, liquid-to-solid ratio, sonication time, and extraction temperature, were investigated by the single-factor experiment. The compositional characterization of flavonoids was characterized with HPLC-UV. Scanning electron microscopy (SEM) was applied to research the surface morphology of raw material. Furthermore, the bioactivities of the extract obtained by UA-ATPE were studied in vitro. Results: The optimal extraction conditions were as follows: the ethanol content was 26.12% (w/w), the ammonium sulfate content was 20.02% (w/w), the liquid-to-solid ratio was 40 mL/g, the sonication time was 5 min with the ultrasonic power of 250 W, and the operating process was performed at room temperature. Compared with the traditional extraction methods, UA-ATPE exhibited higher extraction efficiency and better extraction selectivity. The DPPH and ABTS radical scavenging tests showed that enriched products possessed strong antioxidant activity. Conclusion: The study confirmed that the developed method of UA-ATPE could be used as an efficient, eco-friendly, and low-consumption method for the extraction and purification of flavonoids from Scutellariae Radix.

Source apportionment of PM2.5 in Montréal, Canada, and health risk assessment for potentially toxic elements

Abstract. Source apportionment of PM2.5 was performed using positive matrix factorization (PMF) based on detailed chemical composition data from 24 h filter samples collected over a 3-month period (August–November 2020) at an urban site in Montréal, a Canadian city with a population of approximately 4×106 people. This source apportionment study, which examined the main contributing sources to PM2.5 using a larger suite of organic molecular markers than other Canadian studies, is the first of its sort in Canada. A focus of this study was on quantifying previously unresolved sources of PM2.5 through the inclusion in the PMF analysis of additional organic molecular markers beyond those measured typically by the Canadian government's National Air Pollution Surveillance Program (NAPS). The organic species included in the PMF model were comprised of six n-alkanes, two fatty acids, one dicarboxylic acid, two biogenic secondary organic aerosol (SOA) tracers, and hopane. Secondary inorganic aerosols (SIAs) and SOAs were the dominant components and constituted 39 % of the measured PM2.5 mass, while the local primary anthropogenic sources, namely traffic exhaust, road dust, industrial, and cooking emissions, contributed 23 %. The chemical transport model GEOS-Chem revealed that ammonium sulfate concentrations in Montréal are strongly influenced by both local sources in Quebec and transboundary input from the United States, with the transboundary input exceeding the local emissions for SOA. Co and Cr(VI) presented an elevated cancer risk, highlighting that more attention should be given to these trace metals, which were associated with industrial emissions by the PMF analysis. Furthermore, the results showed that industrial emissions were minor contributors to the total PM2.5 mass concentration but were the largest contributors to Co and Cr(VI) concentrations. Thus, the health hazards associated with this source cannot be entirely established by the PM2.5 mass concentration alone. This study highlights that, when evaluating air quality in Montréal and other urban regions, the prioritization of sources for mitigation strategies will diverge if one considers total PM2.5 mass concentration or the concentration of individual particulate-bound contaminants. Furthermore, the large transboundary contribution from the United States to total PM2.5 levels suggests that future municipal, provincial, and federal monitoring and regulations would be more effective if they focus on specific high-risk contaminants (e.g., Co and Cr(VI) rather than total PM2.5).