Extraction of functional natural products employing microwave-assisted aqueous two-phase system: application to anthocyanins extraction from mulberry fruits.

Aqueous two-phase extraction (ATPE) has been extensively utilized for the extraction and separation of tiny-molecule substances as a new system (system with short-chain ethanol and inorganic salts). In this study, an innovative method of extracting anthocyanins from mulberry was developed, employing microwave-assisted extraction with ethanol/ammonium sulfate as a biphasic extractant. Response surface methodology (RSM) was utilized to optimize anthocyanin extraction conditions: 39% ethanol (w/w), 13% ammonium sulfate (w/w), and liquid-to-solid ratio of 45:1, microwave duration 3 min, microwave temperature 32 °C, and microwave power 480 Watt (W). High-performance liquid chromatography (HPLC) analysis demonstrated no significant differences in the structure of mulberry anthocyanins before and after MAATPE treatment, furthermore. The extraction behavior of MAATPE was due to hydrogen bonding, according to Fourier transform infrared spectroscopy (FT-IR). Scanning electron microscopy analysis found that MAATPE damaged the cell structure via a microwave enhancement effect, which was more favorable to anthocyanin dissolution than standard extraction methods. The DPPH free radical scavenging rate of mulberry extracts at 0.5 mg/mL was higher than that of vitamin C (96.4 ± 0.76%), and the ABTS free radical scavenging rate (82.52 ± 2.13%) was close to that of vitamin C, indicating that MAATPE-derived mulberry extracts have good antioxidant activity.

Ammonium sulfate supplementation enhances erythromycin biosynthesis by augmenting intracellular metabolism and precursor supply in Saccharopolyspora erythraea.

In this study, the cellular metabolic mechanisms regarding ammonium sulfate supplementation on erythromycin production were investigated by employing targeted metabolomics and metabolic flux analysis. The results suggested that the addition of ammonium sulfate stimulates erythromycin biosynthesis. Targeted metabolomics analysis uncovered that the addition of ammonium sulfate during the late stage of fermentation resulted in an augmented intracellular amino acid metabolism pool, guaranteeing an ample supply of precursors for organic acids and coenzyme A-related compounds. Therefore, adequate precursors facilitated cellular maintenance and erythromycin biosynthesis. Subsequently, an optimal supplementation rate of 0.02 g/L/h was determined. The results exhibited that erythromycin titer (1311.1 µg/mL) and specific production rate (0.008 mmol/gDCW/h) were 101.3% and 41.0% higher than those of the process without ammonium sulfate supplementation, respectively. Moreover, the erythromycin A component proportion increased from 83.2% to 99.5%. Metabolic flux analysis revealed increased metabolic fluxes with the supplementation of three ammonium sulfate rates.

Shortening the sulfur cell cycle by a green approach for bio-production of extracellular metalloid-sulfide nanoparticles.

In the present study, a new approach was introduced regarding the extracellular synthesis of selenium sulfide micro/nano-particles using Saccharomyces cerevisiae in different ammonium sulfate supplementation and in the presence of sodium selenosulfate precursors (S1) and a blend of selenous acid and sodium sulfite (S2). In S1, only cell supernatant exposed to ammonium sulfate was able to reduce sodium selenosulfate. Whereas, in S2, cell supernatant in both pre-conditions of with or without ammonium sulfate (S2 + or S2-) were able to reduce selenous acid and sodium sulfite. Electron microscopy, also indicated that selenium sulfide NPs were successfully synthesized with average size of 288 and 332 nm for S2 + and S2- in SEM and 268 and 305 nm in TEM. Additionally, elemental mapping by energy-dispersive x-ray analysis confirmed the presence of sulfur/selenium elements in the particles in a proportion of 24.50 and 23.31 for S2- and S2 + , respectively. The mass spectrometry indicated the probability of Se2S2, SeS1.1, Se2, Se, SeS5, SeS3, Se3S5/Se5, Se3/SeS5, Se6, Se4/SeS7, Se2.57S5.43/Se2S2 and Se4S/Se2S6 molecules for S2 + and of Se, Se2, Se3S5/Se5, Se6 and Se4 species for S2-. In FTIR spectra, primary (i.e. 1090-1020 and 1650-1580 cm-1) and secondary (1580-1490 cm-1) amine bands duly confirmed the protein corona around the NPs.

Nutrient recovery and valorisation from pig slurry liquid fraction with membrane technologies.

Livestock slurry has been reported to be a potential secondary raw material as it contains macronutrients ­nitrogen, phosphorus and potassium-, which could be valorised as high-quality fertilizers if proper separation and concentration of valuable compounds is performed. In this work, pig slurry liquid fraction was assessed for nutrient recovery and valorisation as fertilizer. Some indicators were used to evaluate the performance of proposed train of technologies within the framework of circular economy. As ammonium and potassium species are highly soluble at the whole pH range, a study based on phosphate speciation at pH from 4 to 8 was assessed to improve the macronutrients recovery from the slurry, resulting in two different treatment trains at acidic and alkaline conditions. The acidic treatment system based on centrifugation, microfiltration and forward osmosis was applied to obtain a nutrient-rich liquid organic fertilizer containing 1.3 % N, 1.3 % P2O5 and 1.5 % K2O. The alkaline path of valorisation was composed by centrifugation and stripping by using membrane contactors to produce an organic solid fertilizer -7.7 % N, 8,0 % P2O5 and 2.3 % K2O-, ammonium sulphate solution -1.4 % N- and irrigation water. In terms of circularity indicators, 45.8 % of the initial water content and <50 % of contained nutrients were recovered - 28.3 % N, 43.5 % P2O5 and 46.6 % K2O - in the acidic treatment resulting in 68.68 g fertilizer per kg of treated slurry. 75.1 % of water was recovered as irrigation water and 80.6 % N, 99.9 % P2O5, 83.4 % K2O was valorised in the alkaline treatment, as 219.60 g fertilizer per kg of treated slurry. Treatment paths at acidic and alkaline conditions yield promising results for nutrients recovery and valorisation as the obtained products (nutrient rich organic fertilizer, solid soil amendment and ammonium sulphate solution) fulfil the European Regulation for fertilizers to be potentially used in crop fields.

Exo-polygalacturonase production enhancement by Piriformospora indica from sugar beet pulp under submerged fermentation using the response surface methodology.

This study proposed a novel and cost-effective approach to enhance and optimize the exo-polygalacturonase from P. indica, a root endophytic fungus. In the current investigation, the impact of ammonium sulfate, sugar beet pulp (SBP), and glucose as variables on induction of exo-polygalacturonase from P. indica was optimized using the central composite design (CCD) of response surface methodology (RSM) under submerged fermentation (SmF). Additionally, determination of the exo-polygalacturonase molecular weight and in situ analysis was performed. The optimal reaction conditions, which resulted in the highest enzyme activity, were observed in the following conditions: ammonium sulfate (4 g/L), SBP (20 g/L), and glucose (60 g/L). Under the optimized condition, the maximum enzyme activity reached 19.4 U/ml (127 U/mg), which increased by 5.84 times compared to non-optimized conditions. The exo-polygalacturonase molecular weight was estimated at 60 KDa. In line with the bioinformatic analysis, the exo-polygalacturonase sequence of P. indica showed similarity with Rhizoctonia solani's and Thanateporus cucumeris. These results indicated that SBP acts as a cheap and suitable inducer of exo-polygalacturonase production by P. indica in submerged cultivation. The outcome of this study will be useful for industries to decrease environmental pollution with cost-effective approaches.

A novel and sustainable approach for biotransformation of phosphogypsum to calcium carbonate using urease producing Lysinibacillus sphaericus strain GUMP2.

Phosphogypsum (CaSO4) is produced as a waste by-product during phosphoric acid production in the fertilizer industry. Only 15% of worldwide phosphogypsum production is recycled, while 85% is stored in the vicinity of factories as huge piles resulting in environmental and health hazards. An extensively studied biotransformation of phosphogypsum to calcium carbonate or calcite (CaCO3) using sulfate reducing bacteria (SRBs) is a prolonged process and results in the formation of extremely hazardous H2S gas. Here we report for the first time a novel approach for biotransformation of phosphogypsum to CaCO3 using urease producing Lysinibacillus sphaericus strain GUMP2. The strain could effectively transform phosphogypsum to crystalline, bead-shaped CaCO3 precipitates. In a batch reactor with the PG loading rate of 60 g/L, 100% biotransformation was observed within seven days. After calcite recovery, the ammonium sulfate formed in the supernatant was recovered by precipitation. Urease-producing L. sphaericus strain GUMP2 could be used to remove the hazardous phosphogypsum from the environment by converting it to the industrially useful CaCO3 and ammonium sulfate, a valuable agricultural fertilizer. This novel and sustainable approach could be a promising solution for the hazardous phosphogypsum in the phosphoric acid industries.

Synchronous Extraction, Antioxidant Activity Evaluation, and Composition Analysis of Carbohydrates and Polyphenols Present in Artichoke Bud.

This study investigated the optimization of ultrasonic-assisted aqueous two-phase synchronous extraction of carbohydrates and polyphenols present in artichoke bud, evaluated their antioxidant activities in vitro, and analyzed the composition of carbohydrates and polyphenols by high-performance liquid chromatography (HPLC). The powder mass, ultrasonic time, ammonium sulfate concentration, and alcohol-water ratio were considered the influencing factors based on the single-factor experiment results, and a dual-response surface model was designed to optimize the synchronous extraction process to extract carbohydrates and polyphenols. The antioxidant activity was evaluated by measuring the scavenging capacity of ABTS+· and DPPH· and the reducing capacity of Fe3+. The optimal process conditions in this study were as follows: the powder mass of 1.4 g, ammonium sulfate concentration of 0.34 g/mL, alcohol-water ratio of 0.4, and ultrasonic time of 43 min. The polyphenol content in artichoke bud was 5.32 ± 0.13 mg/g, and the polysaccharide content was 74.78 ± 0.11 mg/g. An experiment on in vitro antioxidant activity showed that both carbohydrates and polyphenols had strong antioxidant activities, and the antioxidant activity of polyphenols was stronger than that of carbohydrates. The HPLC analysis revealed that the carbohydrates in artichoke bud were mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, and arabinose, and the molar ratio was 10.77:25.22:2.37:15.74:125.39:48.62:34.70. The polyphenols comprised chlorogenic acid, 4-dicaffeoylquinic acid, caffeic acid, 1,3-dicaffeoylqunic acid, isochlorogenic acid B, isochlorogenic acid A, cynarin, and isochlorogenic acid C, and the contents were 0.503, 0.029, 0.022, 0.017, 0.008, 0.162, 1.621, 0.030 mg/g, respectively. This study also showed that the carbohydrates and polyphenols in artichoke bud could be important natural antioxidants, and the composition analysis of HPLC provided directions for their future research. Carbohydrates and polyphenols in artichoke buds can be separated and enriched using the optimized process technology, and it is an effective means of extracting ingredients from plants.

Influence of Nitrogen Fertilizer on the Antioxidative Potential of Basil Varieties (Ocimum basilicum L.).

Total phenolic content (TPC) in extracts of basil depended on the cultivar and type of fertilization used in cultivation. TPC was determined spectrophotometrically with the Folin−Ciocalteu reagent. The antioxidant activity of extracts was analyzed by scavenging of DPPH and ABTS radicals, on the basis of metal chelating ability (MetChel) and ferric reducing antioxidant power (FRAP). The greatest TPC was determined in the purple cultivars­141.35 and 165.44 mg gallic acid/g d.m. for fertilized with ammonium nitrate (NH4NO3) and ammonium sulfate ((NH4)2SO4), respectively. Their extracts had the greatest antioxidant capacity in the majority of the methods used. The results varied depending on the modelling system used. The amount of polyphenols in individual basil cultivars differed significantly (p < 0.05) depending on the fertilization used in the culture. Regarding TPC, DPPH, ABTS, FRAP, and MetChel variables, we observed a significant effect for the applied cultivation. In the case of MetChel factor, lower results of all investigated basil species were observed for cultivation with ammonium sulfate. PCA demonstrated in the present study shows that Sweet and Cinnamon Basil samples cultivated with ammonium nitrate create a separated group. We recommend cultivation with ammonium sulfate fertilizers for these varieties of basil. The high content of phenolic compounds demonstrated in Sweet and Cinnamon Basil cultivated with ammonium sulfate, and thus associated antioxidant activity, indicates that it can constitute a valuable source for bioactive compounds in a balanced diet.

Purification of damson plum polyphenol oxidase by affinity chromatography and investigation of metal effects on enzyme activity.

Polyphenol oxidase (PPO) was firstly purified from damson plum as a high antioxidant source. PPO was treated by 0-80% ammonium sulfate precipitation and dialysis. Characterization results were determined for catechol, 4-methyl catechol, pyrogallol and caffeic acid as 0.05 M/pH: 7.2/25 °C; 0.2 M/pH: 4.5/10 °C; 0.01 M/pH: 6.8/5 °C, and 0.2 M/pH: 8.5/10 °C, respectively. Vmax and KM values were calculated for same substrates as 17,219.97 U/(mL*min) and 11.67 mM; 7309.72 U/(mL*min) and 5 mM; 12,580.12 U/(mL*min) and 3.74 mM; 12,100.41 U/(mL*min) and 6.25 mM, respectively. Catechol gave the highest Vmax value among substrates. Affinity purification was performed by using Sepharose 4B-L-Tyrosine-p-aminobenzoic acid and Sepharose 6B-L-Tyrosine-p-aminobenzoic acid. Single bands were approximately observed at 50 kDa for each affinity sample in SDS-PAGE and Native-PAGE. 93.88 and 10.46 purification-folds were obtained for PPO by reference Sepharose-4B and original Sepharose-6B gels. Metal effects upon PPO activity were also investigated due to the importance of enzymatic browning in foods. Cu+2 activation and Fe+2 inhibition were observed with a final metal concentration of 1 mM at 219.66 and 43.18%, respectively. PPO purification from damson plum by affinity chromatography, its characterization, stability evaluation by statistically, and effects of metal ions on damson plum PPO have not been investigated in the literature.

Performance of conventional and enhanced-efficiency nitrogen fertilizers on potato tuber mineral composition and marketability.

BACKGROUND: Potato is an essential crop for global food security, and its cultivation requires a significant amount of readily available nitrogen (N) to ensure tuber quality. Therefore, managing N with enhanced-efficiency fertilizers becomes a potential strategy to meet the seasonal potato N demand. A 3 site-years (SYs) study was conducted to assess the marketable attributes and mineral composition of table-stock potato in response to N rates and fertilizers urea, ammonium sulfate and ammonium sulfate nitrate (ASN) with nitrification inhibitor dimethylpyrazole phosphate (DMPP). RESULTS: At 75% of recommended N rate (RNR), ammonium sulfate and ASN+DMPP ensured marketable tuber yields equivalent to that observed at 100% of RNR. Urea promoted greater tuber K and Mg concentrations than ammonium sulfate and ASN+DMPP. Although inconsistent across SYs, ASN+DMPP generally reduced starch and reducing sugars contents and increased pulp pH and protein content than other fertilizers. Increasing N rates from 50% up to 75% and 100% of RNR increased marketable tuber yields and protein content, whereas soluble solids increased from 50% to 100% of RNR. Conversely, increasing N rates from zero to 75% of RNR reduced tuber firmness, whereas N application reduced tuber P concentration, regardless of N rates. CONCLUSION: Although ASN+DMPP showed potential for increasing marketable tuber yield and protein content, potatoes receiving ammonium sulfate and ASN+DMPP lowered tuber K and Mg concentrations compared to those receiving urea. Overall, potato tuber quality improvements are N source-specific, demanding strategies under which these fertilizers can ensure/improve tuber nutritional composition along with size quality. © 2021 Society of Chemical Industry.

Phycocyanin, a super functional ingredient from algae; properties, purification characterization, and applications.

Phycocyanins (PCYs) are a group of luxuriant bioactive compounds found in blue-green algae with an estimated global market of about US$250 million within this decade. The multifarious markets of PCYs noted by form (e.g. powder or aqueous forms), by grade (e.g. analytical, cosmetic, or food grades), and by application (such as biomedical, diagnostics, beverages, foods, nutraceuticals and pharmaceuticals), show that the importance of PCYs cannot be undermined. In this comprehensive study, an overview on PCY, its structure, and health-promoting features are diligently discussed. Methods of purification including chromatography, ammonium sulfate precipitation and membrane filtration, as well as characterization and measurement of PCYs are described. PCYs could have many applications in food colorants, fluorescent markers, nanotechnology, nutraceutical and pharmaceutical industries. It is concluded that PCYs offer significant potentials, although more investigations regarding its purity and safety are encouraged.

Enhanced Phytase Production by Bacillus subtilis subsp. subtilis in Solid State Fermentation and its Utility in Improving Food Nutrition.

BACKGROUND: Phytic acid acts as anti-nutritional factor in food and feed ingredients for monogastric animals as they lack phytases. OBJECTIVE: Phytase production by Bacillus subtilis subsp. subtilis JJBS250 was studied in solid-state fermentation and its applicability in dephytinization of food. METHODS: Bacterial culture was grown in solid state fermentation using wheat bran and various culture conditions were optimized using 'One variable at a time' (OVAT) approach. Effects of different substrates (wheat bran, wheat straw, sugarcane bagasse), incubation time (24, 48, 72 and 96 h), incubation temperatures (25, 30, 35 and 40°C), pH (4.0, 5.0, 6.0, 7.0 and 8.0) and moisture content (1:1.5, 1:2.0, 1:2.5 and 1:3) were studied on phytase production. Bacterial phytase was used in dephytinization of food samples. RESULTS: Optimization of phytase production was studied in solid state fermentation (SSF) using 'One variable at a time' (OVAT) approach. Bacillus subtilis subsp. subtilis JJBS250 grew well in various agroresidues in SSF and secreted high enzyme titres using wheat bran at 30°C and pH 5.0 after incubation time of 48 h with substrate to moisture ratio of 1:3. Glucose and ammonium sulphate supplementation to wheat bran further enhanced phytase production in SSF. Optimization of phytase production resulted in 2.4-fold improvement in phytase production in solid state fermentation. The enzyme resulted in dephytinization of wheat and rice flours with concomitant release of inorganic phosphate, reducing sugar and soluble protein. CONCLUSION: Optimization resulted in 2.34-fold enhancement in phytase production by bacterial culture that showed dephytinization of food ingredients with concomitant release of nutritional components. Therefore, phytase of B. subtilis subsp. subtilis JJBS250 could find application in improving nutritional quality of food and feed of monogastric animals.

Extraction, Partial Purification and Characterization of Bromelain from Pineapple (Ananas Comosus) Crown, Core and Peel Waste

Abstract Ananas Comosus (also known as pineapple) is a part of Bromeliaceae family and it is consumed as food as well as folk medicine for the treatment of various diseases. It is reported that pineapple is a rich source of bromelain, a cysteine protease and it is considered as an important enzyme in different industries due to its significant therapeutic and industrial applications such as anticancer, anti-inflammatory and meat tenderizing. Bromelain is mostly present in fruit and stem of pineapple, but it is reported that crown, core, and peels, which constitute the waste of the pineapple plant, also contain bromelain but limited data is available. Therefore, the proposed study aimed at utilizing pineapple waste for the extraction and characterization of bromelain. Firstly, crude bromelain was extracted with phosphate buffer (pH 7), then it was subjected to partial purification using different fractions of ammonium sulphate (NH4)2SO4 such as 30, 40, 50 and 60% followed by desalting and concentration. Enzyme activity was calculated by using casein digesting unit (CDU) method. The results demonstrated that the crown bromelain showed highest purification of 4.34-fold at 30% (NH4)2SO4 saturation, whereas core and peel bromelain showed highest purification of 2.75 and 2.59-fold at 40% (NH4)2SO4 saturation. The molecular weight of crude and partially purified bromelain was determined by SDS-PAGE analysis and found to be 26 KDa. The pH and thermal stability of all the parts of pineapple showed maximum stability at pH 7 and at 35oC temperature.

Integration of Nitrogen and Biofertilizer on the Agronomic Efficiency and Quality of Shallot (Allium ascalonicum L.) Plant in the Rainy Season.

BACKGROUND AND OBJECTIVE: In the rainy season farmers don't interest to cultivate shallot because in addition to providing a high dosage of fertilizer they are also sensitive to pathogenic attacks so they are afraid of crop failure and cause low shallot production. This study aimed to knew effect of agronomic component and quality of shallot under different concentrations of biofertilizer and Ammonium Sulphate (AS) fertilizer dose in the rainy season. MATERIALS AND METHODS: The study was conducted in Cangkring, Srandakan, Bantul, Special Region of Yogyakarta Indonesia from August to October 2019. The study was arranged in RCBD factorial with three replications. The first factor was a various dose of ammonium sulphate (100, 200 and 300 kg ha-1). The second factor was various concentrations of biofertilizer (2, 3 and 4%), and control. The observed variables were the analysis of growth yield and quality component of shallot plant. The analyzed using analysis of variance at 5% of significance then continued by DMRT at 5% of significance. RESULTS: There was the interaction between the application of AS dosage and biofertilizer concentration on all of variable observations. There was a significant difference between treatment with control on all of the observation variables. CONCLUSION: The combination of AS fertilizer 200 kg ha-1 dose and 3% biofertilizer concentration increased agronomic efficiency, growth, bulbs yields, and quality of bulbs include provitamin A, oleoresin compounds.

The Quality of Ciders Depends on the Must Supplementation with Mineral Salts.

Providing yeast with the right amount of mineral salts before fermentation can contribute to improving the entire technological process, resulting in a better-quality final product. The aim of this study was to assess the impact of apple must supplementation with mineral salts ((NH4)2SO4, MgSO4, (NH4)3PO4)) on enological parameters, antioxidant activity, total polyphenol content, and the profile of volatile cider compounds fermented with various yeast strains. Rubin cultivar must was inoculated with wine, cider, and distillery or wild yeast strains. Various mineral salts and their mixtures were introduced into the must in doses from 0.167 g/L to 0.5 g/L. The control sample consisted of ciders with no added mineral salts. The basic enological parameters, antioxidant properties, total polyphenol content, and their profile, as well as the composition of volatile compounds, were assessed in ciders. Must supplementation with magnesium salts significantly influenced the use of the analyzed element by yeast cells and was dependent on the yeast strain. In supplemented samples, a decrease in alcohol concentration and total acidity, as well as an increase in the content of extract and total polyphenols, was observed compared to the controls. The addition of ammonium salts caused a decrease in the amount of higher alcohols and magnesium salts, as well as a decrease in the concentration of some esters in ciders.

Extraction, partial purification and characterization of proteases from the red seaweed Gracilaria edulis with similar cleavage sites on κ-casein as calf rennet.

Enzymes currently used in cheesemaking have various drawbacks, and there is a continual need to find new coagulants. This study describes the extraction and biochemical characterization of two proteases from the red alga Gracilaria edulis. The proteases were extracted with phosphate buffer and partially purified by ammonium sulphate precipitation and dialysis. The enzymes exhibited optimum caseinolytic activity at 60 °C and a pH range of 6-8. They showed a high ratio of milk-clotting over caseinolytic activity, indicating they had an excellent milk-clotting ability. The proteases were confirmed to be serine protease and metalloprotease with molecular weight (MW) of 44 and 108 kDa. They exhibited high hydrolytic activity on κ-caseins, cleaving κ-casein at four main sites, one of which being the same as that of calf rennet, which is the first reported for an algal protease. The findings demonstrated that the proteases could potentially be used as a milk coagulant in cheesemaking.

Effect of inorganic salt on partition of high-polarity parishins in two-phase solvent systems and separation by high-speed counter-current chromatography from Gastrodia elata Blume.

Parishins are high-polarity and major bioactive constituents in Gastrodia elata Blume. In this study, the effect of several inorganic salts on the partition of parishins in two-phase solvent systems was investigated. Adding ammonium sulfate, which has a higher solubility in water, was found to significantly promote the partition of parishins in the upper organic polar solvents. Based on the results, a two-phase solvent system composed of butyl alcohol/acetonitrile/near-saturated ammonium sulfate solution/water (1.5:0.5:1.2:1, v/v/v/v) was used for the purification of parishins by high-speed counter-current chromatography. Fractions obtained from high-speed counter-current chromatography were subjected to semi-preparative high-performance liquid chromatography to remove salt and impurities. As a result, parishin E (6.0 mg), parishin B (7.8 mg), parishin C (3.2 mg), gastrodin (15.3 mg), and parishin A (7.3 mg) were isolated from water extract of Gastrodia elata Blume (400 mg). These results demonstrated that adding inorganic salt that has high solubility in water to the two-phase solvent system in high-speed counter-current chromatography was a suitable approach for the purification of high-polarity compounds.

Simultaneous extraction and enrichment of polyphenol and lutein from marigold (Tagetes erecta L.) flower by an enzyme-assisted ethanol/ammonium sulfate system.

Enzyme-assisted aqueous two-phase extraction (EA-ATPE) using ethanol/ammonium sulfate system was investigated for total polyphenol (TP) and lutein from marigold flowers. The important factors were investigated by single factor experiment and response surface methodology combined with Box-Behnken design to optimize the operating parameters of EA-ATPE. The maximum yields of TP and lutein were 83.56 ± 0.69 mg g-1 and 5.59 ± 0.13 mg g-1, respectively. Compared with aqueous two-phase extraction and Soxhlet extraction (SE), the extraction yield of TP by EA-ATPE is 64.91% higher and the extract of EA-ATPE has better antioxidant activity. The pretreatment effect was also researched by scanning electron microscopy. Thus, EA-ATPE is an efficient method for extracting bioactive components from plants.

Protective Effects of ß-Carotene Against Ammonium Sulfate Toxicity: Biochemical and Histopathological Approach in Mice Model.

In this study, the protective role of ß-carotene against ammonium sulfate-induced toxicity has been evaluated in Mus musculus var. albino mice, along with biochemical and histopathological parameters. Some biochemical parameters such as aspartate transaminase (AST), alanine transaminase (ALT), blood urea nitrogen (BUN), creatinine and oxidative stress parameters, malondialdehyde (MDA), and glutathione (GSH) levels in kidney and liver tissues were investigated. The mice were randomly divided into six groups. Group I received intraperitoneal injections of 0.9% NaCl; group II received orally administered 250 mg kg-1 bw ß-carotene, group III received orally administered 500 mg kg-1 bw ß-carotene; group IV received 320 mg kg-1 bw ammonium sulfate; group V was given 250 mg kg-1 bw ß-carotene +320 mg kg-1 of bw ammonium sulfate; and group VI received orally administered 500 mg kg-1 of bw ß-carotene +320 mg kg-1 of bw ammonium sulfate. As a result, it was determined that the ammonium sulfate treatment causes significant changes in the biochemical and oxidative stress parameters and also in histological examinations. In group IV, significant increases in ALT, AST, BUN, MDA, and creatinine levels, and a significant decrease in GSH levels were observed compared with control group. In histopathological examinations, different pathological findings such as proteinaceous deposits, thickening of basement membrane, hyaline cast in kidney tissue and stellate cell, karyomegaly, and binucleated cells in liver tissue were observed. ß-carotene treatment in group V and VI ameliorated the elevated levels of liver enzymes and improved oxidative stress and histopathological findings, and so, it could be concluded that ß-carotene offered remarkable protection against ammonium sulfate-induced toxicity.

Purification and properties of Manganese Superoxide Dismutase (MnSOD) from Tamarix aphylla L.

Superoxide dismutase (SOD) of the Tamarix aphylla leaves were detected at optimum conditions that collected in April, May and June. Results indicated the specific activity in the crude extract reaching to 36.76 unit/ mg protein. Crude SOD was purified by several techniques, precipitation with ammonium sulfate (50-75) %, Ion exchange chromatography using DEAE-cellulose and two steps of size exclusion chromatography on sephacryl S-200 column. The obtained specific activity (310 unit/mg protein) and purification fold 7.91. The purified enzyme revealed one band by SDS-polyacrylamide gel electrophoresis with molecular mass 85.703 kDa. while 89.125 kDa by Sephacryl S-200. The optimal pH and temperature for enzyme activity were 7.5, and 50ºC respectively. EDTA, SDS and NaN3 reduced activity, contrariwise of H2O2 and KCN, pointed to the studied SOD is MnSOD. Michalis constant Km and maximum velocity Vmax values were 0.016 mM and 55.86 mM/min, respectively by using Pyrogallol as substrate. According to the results, we conclude Tamarix aphylla produce MnSOD which can have purified by serial purification techniques for better activity and characterized for further studies.