Ammonium Sulfate Concentration Research Articles

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).

A methodical optimization of melamine and ammonium sulphate ratio for the formation of graphitic carbon nitride nanosheets

A facile method for the preparation of graphitic carbon nitride nanosheets (GCN NS) was reported by adjusting the melamine and ammonium sulphate ratio via 1:1, 1:1.25, 1:1.5, and 1:2 respectively. The bulk sheet with an average thickness of 112 nm was obtained in the absence of ammonium sulphate. Meanwhile, the nanosheets with 34 nm thickness was formed with a higher concentration of ammonium sulphate. The gases like hydrogen, ammonia, ammonium bisulphate, sulfur oxide, etc were generated at high temperature (550 °C) accelerate the exfoliation rate and lead to the formation of nanosheets. A significant electronic and crystallographic changes was observed in UV and XRD analysis, while introducing a higher concentration of ammonium sulphate. The GCN NS obtained at a 1:2 ratio has a band gap energy value of 2.77 eV, which is a more suitable material for photo and electrocatalytic applications.

Rhodotorula paludigena CM33 cultivation process development for high β-carotene single cell protein production

Rhodotorula yeast has potential in terms of biotechnology for the production of various compounds for the environmental, industrial, and medical sectors. It is well known to generate carotenoid pigments that are beneficially distinguished. This investigation aimed to optimize the growth of Rhodotorula paludigena CM33 and its production of β-carotene. Response Surface Methodology was employed to optimize temperature, ammonium sulfate, and ferric chloride concentrations in a 5L-bioreactor batch culture. The optimal conditions of 30 °C, 6.2 g/L of ammonium sulfate, and 75.0 mg/L of ferric chloride provided biomass of 17.71 g/L, a specific growth rate of 0.22 h−1, and β-carotene concentration of 40.29 mg/L. Upon scaling up the cultivation process, the 22L-bioreactor displayed similar growth patterns to those of the 5L-bioreactor. However, the 22L-bioreactor exhibited higher product yields in a shorter time period. The maximum biomass production of 18.96 ± 1.31 g/L and the maximum β-carotene production of 42.81 ± 0.61 mg/L were achieved at 28 h. This scaling-up strategy resulted in improved production efficiency. Additionally, the nutritional value of the cells was evaluated, suggesting their potential as animal feed supplements. R. paludigena CM33 demonstrated a β-carotene single-cell protein content, making it a valuable source for aquaculture and livestock feed. These findings highlight the potential of this yeast strain for large-scale production of β-carotene and single-cell protein, offering nutritional benefits for animal feed.

Purification and Characterization of Alkaline Protease Isolated from Cotton (Gossypium hirsutum) Seeds

Proteases are widely utilized both in physiological and commercial fields such as medicine, food, detergent, and leather. Plant-originated proteases play a significant role in several biomedical fields due to their easy accessibility and activity. Pakistan is an agro-based country and can be an ideal place for the isolation of industrially important proteases from plant sources such as cotton, which is the main crop and frequently available and low cost. Purification of protease was carried out by fractionation with two-fold acetone, ethanol, methanol and various concentrations (40-80%) of ammonium sulphate. The precipitates formed were collected after centrifugation and dialyzed for 24 hours against universal buffer pH 7.0 and was centrifuged in a cooled refrigerated. The dialyzed sample was loaded on Sephadex G–100 gel column. The fractions of the samples were collected and their absorbance of protein was monitored at 280 nm. The homogeneity of the purified enzyme was checked by SDS gel electrophoresis The purified protease enzyme has optimum activity at 30°C and pH 8.0 when casein was used as substrate. The Km and Vmax values of purified cotton seed's alkaline protease activity was recorded as 0.03M and 17 μmol/minute respectively. Protease activity was increased by the addition of cysteine but inhibited by Iodoacetic acid and β-Mercaptoethanol and decreased with some metal ions. These characteristics of the purified enzyme allowed classifying it as a cysteine protease. In conclusion, this study suggests that the alkaline protease enzyme is the best choice for commercial use

Influence of Ammonium Sulfate and Ammonium Nitrate on the Properties of Ambient PM2.5 in Zhenjiang

Recently, the contribution of inorganic salts (nitrates in particular) to the mass concentration of particulate matter with an aerodynamic diameter of less than 2.5 μm (PM2.5) has been increasing across China. However, it is urgent to understand how the increased inorganic salts affect the crucial properties of PM2.5. Here, we conducted continuous field observations at Zhenjiang Ecology and Environment Protection Bureau from January 1 to December 31, 2021. The mass concentrations of ammonium sulfate[(NH4)2SO4] and ammonium nitrate (NH4NO3) were calculated using different methods. The contributions of (NH4)2SO4 and NH4NO3 to the extinction coefficient, hygroscopic growth, and acidity of PM2.5 were discussed in detail. Our results demonstrated that the mean mass concentrations of (NH4)2SO4 and NH4NO3 during the study period were (6.5±4.5) and (15.0±13.3) μg·m-3, which contributed (20.5±18.2)% and (34.5±18.4)% to the mass concentration of PM2.5, respectively. The total extinction coefficient of PM2.5 was (224.5±194.2) Mm-1, in which NH4NO3 was the largest contributor[(40.1±20.9)%] followed by (NH4)2SO4[(19.1±10.8)%]. (NH4)2SO4 and NH4NO3 were also the dominant contributors to the hygroscopic growth of PM2.5. In particular, NH4NO3contributed from (53.8±13.4)% to (61.6±14.6)% to the aerosol water content of PM2.5 under pollution conditions. Thus, NH4NO3 was a key air pollutant to be targeted for further improving the visibility and air quality in Zhenjiang in the future. However, the reduction in the precursors of NH4NO3 would lead to an increase in aerosol acidity, particularly in the spring and winter seasons. Our results help us understand the evolution of air quality and the related impacts and also provide important information on air quality improvement in Zhenjiang in the future.

In vitro α-Amylase and α-Glucosidase Inhibitory Activity of Dioscorin from Dioscorea alata

Dioscorea alata is a species of yam found in many countries in Southeast Asia. It contains many bioactive compounds consisting of steroidal saponins, phenolic compounds flavonoids, polysaccharides, and proteins. Eighty to eighty-five of the soluble proteins in the tuber of D. alata are dioscorin. Dioscorin (31 kDa) is the major storage protein of yam tuber, exhibiting biochemical and immunomodulatory properties, such as antioxidant activity, reducing blood pressure and angiotensin I converting enzyme (ACE) inhibitors activity. The aims of this research focused on extraction of the total soluble protein from D. alata tuber, purification of dioscorin, and study of in vitro inhibitory activity of α-amylase and α-glucosidase of dioscorin extracts. The results showed that a maximum yield of total soluble protein was obtained from the buffers extraction using Tris-HCl buffer at pH 9.0 resulting in the protein concentration of 15.10 mg/g with a protein extraction yield of 31.56 ± 2.36%. The crude protein was further recovered using ammonium sulfate precipitation and dialysis. SDS-PAGE analysis showed that dioscorin from crude extracts was precipitated at the concentration of ammonium sulfate of 60% saturation. The purified dioscorin was obtained from anion exchange chromatography (DEAE-Sephacel) and its molecular weight of 31 kDa was estimated on SDS-PAGE. Furthermore, the α-amylase inhibitory activities of crude extract (1 mg/mL), purified dioscorin obtain from gastric digestion and purified dioscorin obtained from intestinal digestion were at 14.44% 2.21% and 78.13%, respectively. Moreover, the results showed that the α-glucosidase inhibitory activity of the crude extract, and purified dioscorin obtained from gastric digestion were at 10.56% and 2.61%, respectively.

Kinetic isotope effect study of N-6 methyladenosine chemical demethylation in bicarbonate-activated peroxide system.

N-6 methyladenosine is the most abundant nucleic acid modification in eukaryotes and plays a crucial role in gene regulation. The AlkB family of alpha-ketoglutarate-dependent dioxygenases is responsible for nucleic acid demethylation. Recent studies have discovered that a chemical demethylation system using hydrogen peroxide and ammonium bicarbonate can effectively demethylate nucleic acids. The addition of ferrous ammonium sulfate boosts the oxidation rate by forming a Fenton reagent with hydrogen peroxide. However, the specific mechanism and key steps of this process remain unclear. In this study, we investigate the influence of ferrous ammonium sulfate concentration on the kinetic isotope effect (KIE) of the chemical demethylation system using LC-MS. As the concentration of ferrous ions increases, the observed KIE decreases from 1.377 ± 0.020 to 1.120 ± 0.016, indicating a combination of the primary isotope effect and inverse α-secondary isotope effect with the ion pairing effect. We propose that the initial hydrogen extraction is the rate-limiting step and observe a tight transition state structure in the formation of the hm6A process through the analysis of KIE trends. The concentration-dependent KIE provides a novel perspective on the mechanism of chemical demethylation and offers a chemical model for enzyme-catalyzed demethylation.

Effect of carbon and nitrogen concentrations on lipid accumulation and regulation of acetyl-CoA carboxylase in Papiliotrema laurentii.

Biodiesel is an interesting alternative to petroleum diesel as it is renewable, biodegradable, and has a low pollutant content. Yeast oils can be used for biodiesel production instead of edible oils, mitigating the use of arable land and water for biodiesel production. Maximum lipid accumulation is reached at 48h of cultivation by the oleaginous yeast Papiliotrema laurentii UFV-1. Nevertheless, the effects of carbon and nitrogen concentrations on lipid accumulation, as well as the regulation of lipid metabolism in this yeast are still not well-characterised. Therefore, this work evaluated the effects of carbon and nitrogen concentrations on the lipid accumulation in P. laurentti, the expression of the ACC gene, and the activity of the enzyme acetyl-CoA carboxylase (ACCase) in different carbon:nitrogen ratios (C:N) and glucose concentrations. The variation of ammonium sulfate concentration did not affect the growth and lipid accumulation in P. laurentii UFV-1. On the other hand, glucose concentration remarkably influenced biomass and lipid production by this yeast. Therefore, the carbon concentration is more important than the nitrogen concentration for lipid production by P. laurentii UFV-1. Importantly, the levels of both ACC gene expression and ACCase activity were maximum during the late-exponential growth phase and decreased after reaching the highest lipid contents, which was easier evidenced during the accumulation and maximum lipid levels. As such, the reduction of ACCase enzyme activity seems to be related to the decrease in the expression level of the ACC gene.

Curcuminoid extraction from rhizomes of Curcuma longa via aqueous two phases system

The extraction of curcuminoids [curcumin (C21H20O6, CUR), demethoxycurcumin (C20H18O5, DMC), and bisdemethoxycurcumin (C19H16O4, BDMC)] from Curcuma longa rhizomes was investigated using an aqueous two-phase system (ATPS) with ethanol/salt. The optimized conditions for the extraction process were determined by a central composite design to be an ethanol concentration of 13.6%, an ammonium sulfate concentration of 33.2%, and a temperature of 66.8 °C. Under these conditions, the highest yield of 97.9% was achieved, accompanied by a notable antioxidant activity of 84.38%. The experimental data exhibited good agreement with a second-order kinetic model, as confirmed by the evaluation of the coefficient of determination (R 2), normalized standard deviation, and average relative error.

A monolithic stationary phase with dendritic nanostructures for the separation of PEGylated proteins.

Separation of PEGylated protein mixtures into individual species is a challenging procedure, and many efforts have been focused on creating novel chromatographic supports for this purpose. In this study, a new monolithic stationary phase with hyperbranched nanostructures was chemically synthesized. For this, monoliths with a support matrix of poly (glycidyl methacrylate-co-ethylene dimethacrylate) and ethylenediamine chemistry were modified with third-generation dendrons with butyl-end groups. The new monolith was analyzed by infrared spectroscopy, confirming the dendron with butyl ligands and exhibited low mass transfer resistance as observed by breakthrough frontal analysis. This support was able to separate mono-PEG ribonuclease A from the PEGylation mixture, indicated by a single band (∼30kDa) in the electrophoretic analysis. Moreover, the separation of mono-PEGylated positional isomers was probably observed, as the protein with ∼30kDa was found in two separate peaks. Interestingly, the dendronized monolith allowed the separation of the reaction mixture into individual PEGylated species when using high ammonium sulfate concentrations (2M). A correlation between the PEGylation degree and the strength of the hydrophobic interactions on the monolith was observed. This chromatographic approach combines the natural branched architecture of dendrons and the higher capabilities of the monoliths enhancing the hydrophobic surface area, and therefore the interaction between the PEGylated proteins and ligands. Thus, the novel support represents a novel platform for the purification of PEGylated from non-PEGylated proteins with biotechnological applications.

Production of Industrial Applicable Enzymes from Rhizopus Oryzae through Response Surface Methodology

The agricultural wastes are used for production of vital enzymes like cellulase, protease and lipase through Solid State Fermentation (SSF), which reveal their industrial potential aspect. This study was done to find out the ability of Rhizopus oryzae to produce lipase, protease and cellulase through SSF. Wheat bran showed maximum enzyme activities as related to others agricultural wastes. The enzymes showed maximum activities (21.4±0.04) (4.6±0.02 IU/mL) and (1.1 ± 0.01 IU/mL) for cellulase, lipase and protease respectively at temperature 25ºC and incubation period 72 hours. Response surface methodology (RSM) used to optimize conditions likes temperature, pH, moisture contents, inoculum size and substrate concentration. RSM revealed maximum enzyme activities using 100% moisture contents, temperature 25ºC, pH 5, substrate concentrations 12g and inoculum size of 6 mL. The method of ammonia sulphate precipitation used for the purification and crystallization of enzymes like lipase, protease and cellulase. The precipitation of cellulase, lipase and protease performed with 80%, 70 % and 90% ammonium sulphate concentrations to increase yield. The characterization of lipase, protease, cellulase revealed that it worked best below acidic conditions (pH 5) and at moderate temperature (25ºC). The results revealed that the mold oryzae had potential to produce industrially important enzymes at acidic ph.

Investigating the performance of susceptibility-weighted images in Fricke gel dosimeter reading and optimizing the related imaging parameters

Fricke gel dosimeters are especially useful in small-field dosimetry and validation of treatment delivery in three-dimensional space with features such as tissue equivalence, non-toxicity, high spatial resolution, non-dependence on energy, and dose rate. The use of basic Magnetic Resonance Imaging (MRI) protocols (T1- and T2-Weighted) for reading Fricke gel dosimeters has always been considered the dominant method in many studies. However, the development and application of advanced MRI techniques for more accurate readings of Fricke gel dosimeters can be useful. Considering that in the main structure of Fricke gel, there are conversions of iron ions to each other, this study aimed to investigate the performance of Susceptibility-Weighted Imaging (SWI) and Quantitative Susceptibility Mapping (QSM) based on magnetic susceptibility in the reading of Fricke gel dosimeters and to optimize the related imaging parameters. For this purpose, a Fricke-Xylenol orange-gelatin was made at five concentrations of iron ammonium sulfate. To obtain gel dosimeter calibration curves, vials containing gel were subjected to irradiation at three different doses by a linear accelerator. The reading of gel dosimeters was performed using MRI imaging in three protocols, T1W, T2W, and SWI, and analyzed with a method unique to each one. Finally, the results obtained from the three protocols were compared with each other. The comparison of calibration curves in three imaging protocols shows that the sensitivity of calibration curves in SWI was about three times its value in T2W, and on the other hand, the reported sensitivity in T1W was very small compared to the other two protocols. The linearity factor was similar between SWI and T1W protocols and higher in T2W. Therefore, it is concluded that in addition to the relaxometry techniques that have been used as a conventional method for reading Fricke gel dosimeter, SWI imaging has high sensitivity and specificity for reading dosimeter gel based on iron.