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The determination of inorganic phosphate in the presence of labile phosphate
... The reaction mixture was incubated at 37 C for 30 min and stopped by the addition of 0.1 ml of 50% TCA. Samples were then assayed for inorganic phosphate using the method of Lowry and Lopez (1946) as modified by Phillips and Hayes (1977). 29,30 The colour was read at 800 nm in a spectrophotometer. ...
... Samples were then assayed for inorganic phosphate using the method of Lowry and Lopez (1946) as modified by Phillips and Hayes (1977). 29,30 The colour was read at 800 nm in a spectrophotometer. Mg 2þ -ATPase activity was measured in presence of 1 mM ouabain, a specific inhibitor of Na þ , K þ -ATPase (McIlwain, 1963). ...
... Phillips and Hayes (1977). 29,30 Mg 2þ -ATPase activity was measured in the presence of 0.5 mM EGTA and this value was subtracted from total ATPase activity to get Ca 2þ -ATPase activity. Enzyme activity was expressed as m moles of inorganic phosphate formed/mg protein/hr ...
Epilepsy is a chronic central nervous system disorder that occurs not only with the imbalance of glutamatergic neurons and inhibitory gamma-aminobutyric acid (γ-GABA) neurons, but also with abnormal Central cholinergic neuronal regulation. Since long term usage of antiepileptic drugs cause high incidence of pharmacoresistance and untoward side effects, attention has been paid in recent years to screen bioactive compounds from natural medicinal plants for the treatment of several neurological disorders including Epilepsy. Keeping in view of relative importance of natural medicinal plants, the present study is mainly focused to characterize the anti-convulsant effect of Bacopa monnieri (BM), an Indian herb which is being extensively used in Ayurvedic treatments related to neurological complications. The present study is designed to assess the neurotoxicity of Pentylene tetrazole (PTZ), an epileptic compound with particular reference to Cholinergic system and ATPases in different brain regions of rat to explore the possible antiepileptic effect of different extracts of BM in comparison with Diazepam (DZ) (Reference control). The activity levels of Acetyl cholinesterase (AChE) and ATPases were decreased in different regions of brain during PTZ induced epilepsy which were increased in epileptic rats pretreated with different extracts of Bacopa monnieri except EAE and AE. In addition Acetylcholine (ACh), levels were increased during PTZ induced epilepsy when compared with normal control and levels were reversed on pretreatment with different extracts of BM. Recoveries of these parameters suggest that the bioactive factors present in the extracts offer neuroprotection by interrupting the pathological cascade that occurs during epileptogenesis.
... Further, majority of phosphate-estimating protocols, including the official methods followed by the American Oil Chemists' Society (AOCS) employs bulky glassware, large sample sizes and preparation volumes (Bell and Doisy 1920;Fiske and Subbarow 1925;Han 1995;Taylor and Miller 1914). It has so far been authentically demonstrated by several dynamic researchers that (i) ammonium molybdate is the key for determination of phosphate; (ii) molybdate forms phosphomolybdic acid on reaction with phosphate under acidic conditions; (iii) molybdenum reacts with phosphate in the ratio of 12:1; (iv) oxidised state of phosphomolybdic acid is yellow, while it turns blue on reduction; and (v) a reducing agent is invariably required to reduce oxidised phosphomolybdic acid into reduced phosphomolybdic acid (Baginski et al. 1975;Bell and Doisy 1920;Berenblum and Chain 1938;Briggs 1922;Fiske and Subbarow 1925;Ganesh et al. 2012;Lowry and Lopez 1946;Murphy and Riley 1962;Nagul et al. 2015a, b;Taylor and Miller 1914;Worsfold et al. 2016;Wu 1920). ...
... One of the major drawbacks of the prevailing spectrophotometric phosphate quantification methods has been the failure to curb reduction of molybdic acid without hampering the reduction of phosphomolybdic acid by the reductants used till date (Berenblum and Chain 1938;Han 1995;Lowry and Lopez 1946;Nagul et al. 2015a, b). We initially made attempts to identify and optimise the conditions that could resolve this serious issue. ...
... We preferred to use ascorbic acid (Chen et al. 1956;Paliwal 1967, 1969;Murphy and Riley 1962) over other commonly used reductants such as stannous chloride (Levine et al. 1955;Nagul et al. 2015a), hydrazine sulphate (Boltz and Mellon 1947;Katewa and Katyare 2003;Mendham et al. 2002) and aminonaphtholsulfonic acid (Fiske and Subbarow 1925;Griswold et al. 1951), as this reductant is eco-friendly, natural bio-molecule that is invariably used as an antioxidant by all living beings, right from prokaryotes to eukaryotes with no known negative impact on cellular metabolism or the health of living beings (including humans). As both molybdic acid as well as phosphomolybdic acid (which is yellow in oxidised state) turn blue on reduction, the socalled molybdenum blue would most probably be composed of both reduced molybdic acid as well as reduced phosphomolybdic acid (Baginski et al. 1975;Crouch and Malmstadt 1967;Davies et al. 1973;Lowry and Lopez 1946). The development of molybdenum blue complex by the reductant and its stability relies on (i) H + concentration, (ii) concentration of ammonium molybdate and (iii) concentration of ascorbic acid besides duration of heating or incubation at 100°C. ...
Owing to its essentiality for cellular metabolism, phosphate (PO4³⁻) plays a pivotal role in ecosystem dynamics. Frequent testing of phosphate levels is necessary to monitor ecosystem health. Present investigations were aimed to identify the key factors that are essential for proper quantification of PO4³⁻. Primarily, H⁺ levels played a critical role in the development of molybdenum blue complex by ammonium molybdate and PO4³⁻ with ascorbic acid as a reductant. Molybdenum blue complex formed in the presence of 8 to 12 mmol of H⁺ in 3 ml reaction mixture remained stable even after 72 h. Of different concentrations of ammonium molybdate and ascorbic acid tested, best molybdenum blue complex was formed when their concentrations were 24.3 and 5.68 μmol, respectively. More or less similar intensity of molybdenum blue complex (due to reduction of phosphomolybdic acid and not molybdic acid) was formed in the presence of H⁺ at levels ranging from 8 to 10 mmol in 3 ml reaction mixture. Our findings unequivocally demonstrated that (i) the reaction mixture containing 3% ammonium molybdate, 0.1% ascorbic acid and 5 M H2SO4 in the ratio of 1:1:1 is ideal for PO4³⁻ quantification; (ii) antimony (Sb) significantly curbs the formation of molybdenum blue under these ideal conditions; (iii) this fine-tuned protocol for PO4³⁻ quantification could be extended without any problem for determining the level of PO4³⁻ both in plant as well as soil samples; and (iv) Azotobacter possesses potential to enhance levels of total PO4³⁻ in leaves and grains and soluble/active PO4³⁻ in rhizosphere soils of wheat.
... For digestibility, the ADC of nutrients was calculated based on ratio of amount of chromium oxide to nutrients in the diet and feces. Phosphorus was analyzed using a UV-VIS spectrophotometer (UV-2550, Shimadzu) as described by Lowry and Lopez (1946) [27]. Chromic oxide concentration was determined after acid digestion [28] using a UV-VIS spectrophotometer. ...
... For digestibility, the ADC of nutrients was calculated based on ratio of amount of chromium oxide to nutrients in the diet and feces. Phosphorus was analyzed using a UV-VIS spectrophotometer (UV-2550, Shimadzu) as described by Lowry and Lopez (1946) [27]. Chromic oxide concentration was determined after acid digestion [28] using a UV-VIS spectrophotometer. ...
This study investigated the effect of the inclusion of an enzyme complex-treated rice protein concentrate (RPC) in an extruded diet of juvenile rainbow trout (Oncorhynchus mykiss). A mixture of RPC, corn gluten and soybean meal was pretreated with an enzyme complex before extrusion processing of the diets. An enzyme complex-pretreated RPC (5, 10 and 15%) was formulated with 20% fish meal. A diet without RPC was used as a control. A total of 240 rainbow trouts with an average body weight of 6.04 g were placed in 12 rectangular glass aquaria and fed one of the experimental diets at apparent satiation for 12 weeks. There were no significant differences in growth, feed intake and survival among the groups. Slightly inferior weight gain and specific growth rate were observed in the RPC15E group compared to those in the other groups. The protein and ash contents of the whole body of the final fish were not significantly different among all groups, but the lipid content was significantly lower than that of the control when the dietary RPC level was >10%. Protein digestibility was negatively affected by increased RPC levels in diet. Significantly lower phosphorus digestibility was observed in fish fed the diet containing 5% enzyme-treated RPC than the control. Although there was no significant difference in lipid digestibility in all groups, significantly lower lipid retention was observed in fish fed a diet formulated with more than 10% RPC. These results suggest that the inclusion of RPC in the diet affects lipid retention and the content of rainbow trout. It also decreased protein digestibility. In conclusion, rainbow trout can be fed an extruded diet formulated with 20% fishmeal and 10% enzyme-treated RPC without negative effects on fish growth.
... Як контроль використовували дистильовану воду. Вміст амонію визначали з використанням реактиву Неслера [7], вміст фосфатівметодом Лоурі -Лопеса [8], вміст нітратівіз реактивом Грісса [9]. Вміст гумінових кислот визначали методом УФ/Візспектроскопії на спектрометрі UVmіnі-1240 Shimadzu (Японія) за калібрувальним графіком (за стандартними розчинами). ...
... Наступним етапом були порівняльні дослідження впливу одержаних органічних добрив (до та після обробки ЕГУ) на морфометричні показники рослин і вміст пігментів фотосинтезу (рис. [5][6][7][8]. ...
Проблематика. Розвиток агропромислового комплексу, інтенсифікація рослинництва спричиняють забруднення ґрунтів залишками добрив, гербіцидів. Тому актуальним завданням є зменшення кількості використовуваних мінеральних добрив, застосування сучасних екологічно безпечних агротехнологій. Серед перспективних способів отримання органічних добрив та обробки ґрунту заслуговує на увагу метод електрогідравлічного удару (ЕГУ). Такий підхід сприяє збагаченню ґрунтів азотом, фосфором, покращенню їх структури та родючості.
Мета. Оцінка впливу ЕГУ на якісний і кількісний склад органічних екстрактів (гною великої рогатої худоби, копролітів дощових черв’яків, курячого посліду) та впливу отриманих добрив на морфометричні показники проростків кукурудзи і вміст пігментів фотосинтезу.
Методика реалізації. Обробка органічних субстратів ЕГУ проводилась на імпульсному генераторі потужністю 5кВт. Визначення вмісту амонію у добривах – з реактивом Неслера, фосфатів – методом Лоурі–Лопеса, нітратів – з реактивом Грісса, гумінових кислот – УФ/Віз-спектроскопією. Морфометричні показники, вміст пігментів фотосинтезу проростків кукурудзи визначали на 7-му добу.
Результати. Обробка ЕГУ сприяла збільшенню вмісту фосфатів, нітратів (15–60 %), амонію (8–14 %) і гумінових кислот (50–58 %) у добривах. Як наслідок, за дії оброблених добрив із курячого посліду і копролітів підвищувалися морфометричні показники проростків порівняно з варіантами без обробки ЕГУ (маса коренів – на 160–200 %). За впливу усіх органічних добрив після дії ЕГУ підвищувався також вміст пігментів фотосинтезу в рослинах: вміст каротиноїдів зростав на 8,7 % за використання курячого посліду і на 10 % для копролітів.
Висновки. Обробка органічних відходів із гною великої рогатої худоби, копролітів дощових черв’яків та курячого посліду методом ЕГУ є ефективним підходом до одержання екологічно безпечних добрив, які характеризуються високим вмістом фосфатів, нітратів і амонію, що є сприятливим фактором при вирощуванні рослин, зокрема на ранніх стадіях. За дії органічних добрив з екстрактів з курячого посліду і копролітів істотно підвищувалися морфометричні показники проростків кукурудзи щодо контролю. Результати досліджень свідчать про перспективи продовження досліджень ефективності методу ЕГУ в різних галузях, зокрема для очищення і збагачення ґрунтів, обеззараження промислових стоків тощо.
... The percentage of the Zn-ions dissolved in the solution was determined by centrifuging at 3200 g for 20 min and the zinc content in the filtrate suspension was measured using atomic absorption (Nov AA 400, Analytic Jena, Germany). The soluble zinc was calculated by dividing the measured zinc (as Zn) per the total used ZnO-NPs (1 g) and multiplying by 100 (Lowry and Lopez, 1946). General experimental procedures: The marine water rotifer, B. plicatilis, Müller cysts obtained from the Shrimp Research Institute in Bushehr, Iran, and incubated in standard seawater at 20 ppt. ...
... ZnO NPs uptake by rotifers: Freeze dried samples were weighed and wet-digested with nitric acid (65% HNO3 Suprapur®, Merck, Germany) using MLS-1200 MEGA Microwave for 30 minutes under cold water and were then diluted with distilled water to reach the required volume. The zinc content was obtained using an atomic absorption device (Nov AA 400, Analytic Jena, Germainy) (Lowry and Lopez, 1946). Statistical analysis: After ensuring the normalization of the data by Shapiro-Wilk test, the homogeneity of variances was investigated via Levene's test. ...
In the present study, the effects of ZnO nanoparticles (NPs) on marine rotifer, Brachionus plicatilis, was investigated in three separate experiments. Firstly, the sensitivity and reproductive characteristics of B. plicatilis were studied at concentrations of 0, 0.1, 0.5, 1, 3, 5 and 10 mg L-1 of ZnO-NPs for 10 days. Based on the results, the total number of rotifers (TNR) significantly decreased at 5 and 10 mg L-1 of ZnO-NPs. In addition, the specific growth rate (SGR) of animals was negative at two of the concentrations of ZnO-NPs. In the second experiment, the TNR at 4 concentrations of ZnO-NPs (0, 10, 13, 17, and 19 mg L-1) during 72 h were tested and the 24-72 h LC50 of ZnO-NPs was calculated. After three days, the entire population of rotifers was generally lost at 19 mg L-1 of ZnO NPs. The LC50 of ZnO-NPs in animals at 24, 48, and 72 h intervals was registered as 18.2±1.34, 12.43±0.08, and 9.63±0.26 mg L-1 , respectively. Finally, the zinc accumulation in rotifers was measured at different concentrations (0, 0.1, 0.5, and 1.3 mg L-1) of ZnO-NPs and maximum zinc (123 μg g-1 of rotifer DW) uptake by rotifers was observed in treatment 3 mg L-1 of ZnO-NPs. In sum, it can be concluded that the B. plicatilis can be used as a biological model for studying marine water contaminants with nanoparticles, especially ZnO-NPs.
... This P i reacts with the ammonium molybdate to form a phosphomolybdic acid complex which is reduced in the presence of ascorbic acid into a blue molybdous compound that absorbs at 700 nm. This reaction requires an acidic pH [20]. Therefore, ammonium molybdate and ascorbic acid were prepared in H 2 SO 4 at different concentrations to determine the most suitable one for the dosage simultaneously of high and low concentrations of phosphate. ...
... Standard solutions are prepared by serial dilutions of a stock solution of phosphate in tris-HCl (50 mM, pH 8.5) buffer. The P i dosage is determined according to the method of Lowry et al. [20] with minor modifications. Briefly, 100 µL of a 1% solution of ammonium molybdate prepared in H 2 SO 4 is mixed with 1000 µL P i solutions leading to the formation of a phosphomolybdic acid complex. ...
Bacterial resistance has become a worrying problem for human health, especially since certain bacterial strains of Escherichia coli (E. coli) can cause very serious infections. Thus, the search for novel natural inhibitors with new bacterial targets would be crucial to overcome resistance to antibiotics. Here, we evaluate the inhibitory effects of Apis mellifera bee venom (BV-Am) and of its two main components -melittin and phospholipase A2 (PLA2)- on E. coli F1F0-ATPase enzyme, a crucial molecular target for the survival of these bacteria. Thus, we optimized a spectrophotometric method to evaluate the enzymatic activity by quantifying the released phosphate from ATP hydrolysis catalyzed by E. coli F1F0-ATPase. The protocol developed for inhibition assays of this enzyme was validated by two reference inhibitors, thymoquinone (IC50 = 57.5 μM) and quercetin (IC50 = 30 μM). Results showed that BV-Am has a dose-dependent inhibitory effect on E. coli F1F0-ATPase with 50% inhibition at 18.43 ± 0.92 μg/mL. Melittin inhibits this enzyme with IC50 = 9.03 ± 0.27 μM, emphasizing a more inhibitory effect than the two previous reference inhibitors adopted. Likewise, PLA2 inhibits E. coli F1F0-ATPase with a dose-dependent effect (50% inhibition at 2.11 ± 0.11 μg/mL) and its combination with melittin enhanced the inhibition extent of this enzyme. Crude venom and mainly melittin and PLA2, inhibit E. coli F1F0-ATPase and could be considered as important candidates for combating resistant bacteria.
... The ash was treated in sequence with a few ml of nitric and hydrochloric acid, each time expelling the acid on the hot plate. The ash was dissolved in 5 % TCA and filtered; P was measured by the method of Lowry & Lopez (1946). This represented the P bound in the nucleic acids associated with the 'true protein', called protoplasmic P and denoted by PROP. ...
... The TCA extract and washings were collected and made up to 100 ml with cold 5 % TCA, then analysed for free inorganic orthoposphate by the method of Lowry & Lopez (1946). This fraction represents inorganic P, abbreviated to INP. ...
Sunflower plants were alternated between complete nutrient sol. and sol. devoid of either P or N. Successive harvests of top growth were analysed for P and N fractions to follow the time course of depletion and restoration. Under adequate P the TCA-insoluble protoplasmic substance contained 25 mol P/1000 mol N. Unrestrained incorporation of P needed the presence of at least 25 mol inorganic P/1000 mol protoplasmic N; P was adequate with at least 50 mol of total P/1000 mol N converted to proteins and nucleic acids. This ratio agreed with the ratio total P:total N at which max. growth was attained in published work of both field and pot experiments, presumably because most of the N was protein bound. Because of the possibility of higher contents of TCA-soluble N, it is better that total P be related to the TCA-insoluble N. (Abstract retrieved from CAB Abstracts by CABI’s permission)
... After completing digestion process, the mixture was cooled at room temperature and filtered and transferred to a conical flask and 10 ml HNO3 solution (0.1 N) was poured on it. Finally, the digested solution was read using a novAA® 400 P Atomic Absorption (Analytic Jena) with Lowry and Lopez (Lowry and Lopez, 1946). At the end of the experimental trial (50 DAH), 3 larvae were sampled of each replicate (n = 9 larvae per treatment). ...
Selenium (Se) is an essential trace element that has vital effects on growth, antioxidant capacity and early development in marine fish larvae. However, traditional livefoods such as rotifer and artemia lack of Se. In this regard, a 50-day feeding experiment was conducted in order to examine the effects of Rotifer and Artemia enrichment with selenium nanoparticles (SeNPs) on growth, antioxidant capacity and fatty acid profile of Arabian yellowfin seabream (Acanthopagrus arabicus) larvae. Four levels of SeNPs were used to prepare the experimental emulsions,including control (0), 0.5, 1.0 and 5 mg SeNPs L − 1. Larvae were stocked at density of 50 larvae L − 1 into twelve 250-L cylindrical tanks. Tanks supplied with seawater (23 ± 1 ºC; 40.0 ± 1.0 g L − 1). Green water was used from day first until 20 days after hatch (DAH). Rotifers (15 ml − 1) were offered to larvae from day 2-21 DAH and Artemia metanauplii (1-3 Artemia ml − 1) were offered to larvae from day 18-45 DAH. Fish were sampled at 0, 7, 10, 15, 22, 30, 40, 45, 50 DAH. The Se concentration in the livefoods and larval whole body linearly increased with the increment of SeNPs level in the experimental emulsions. Larvae wet weight showed quadratic response and increased with SeNPs up to 1 mg mg SeNPs/L, then decreased in SeNPs (5.0) group. The activity of antioxidant enzymes including catalase, superoxide dismutase glutathione peroxidase, and total antioxidant capacity increased but malondialdehyde level decreased in larvae fed the SeNPs-enriched livefoods and showed both linear and quadratic responses. Lysozyme activity in the whole body of larvae fed SeNPs-enriched livefoods had both linear and quadratic responses and in the most of the sampling times was higher than the control group. Docosahexaenoic acid (DHA) level in the livefoods and larval whole body increased with SeNPs. In conclusion, livefoods enrichment with SeNPs at 1 mg L − 1 is recommended to elevate the antioxidant capacity and non-specific immune response in A. arabicus larvae.
... This would provide a significant improvement in differentiating blood cells. The 5'NT activity was measured in serum according to Fiske and Subbarow's method (Lowry & Lopez, 1946). The principle of this method is based on the estimation of the number of inorganic phosphate micromole resulting from the reduction of 5'-AMP in the interaction medium using Fiske and Subbarow's detector (Goldberg, 1973;Pratibha et al., 2004). ...
One of the hydrolytic enzymes that catalyzes the hydrolysis of 5' -nucleotides into ribonucleosides and phosphate is the 5'nucleotidase, which is found in several organs. Oxidative stress shows depletion of antioxidants, such as glutathione (GSH), and the increased concentration of product of lipid peroxidation, such as MDA. The study comprised a total of 54 subjects including patients with beta thalassemia (n=27) and healthy volunteers (n=27) matched by age and gender. Hematology (Hb and % PCV), 5'-nucleotidase, Ferritin, GSH and MDA were measured using different methods and devices. The 5'nucleotidase, Ferritin, and MDA levels were significantly increased (p<0.001) in beta-thalassemia patients compared to the controls. As opposed to that, the Hb, % PCV and GSH levels were significantly decreased (p<0.001) in patients with beta-thalassemia. There was a negative significant correlation (r = -0.146, -0.245, p<0.05) between Hb and 5'- NT enzyme and PCV% with 5'- NT enzyme, respectively. While Hb had a high significant correlation (r = 0.956, p<0.05) with PCV%. In addition, 5'- NT enzyme had a positive significant correlation with GSH, MDA and Ferritin (r = 0.584, 0.442, 0.735, p<0.05), respectively. The present findings indicated an increase in the level of effectiveness of the 5'nucleotidase enzyme, ferritin and GSH in β thalassemia patients while decreasing in MDA.
... The use of 4-methylumbelliferyl phosphate (Figure 2(c)) and a-naphthyl phosphate as substrates have allowed for the development of fluorogenic tests with extremely high sensitivity 6,42 . The product of enzymatic hydrolysis of a-naphthyl phosphate, i.e. a-naphthol forms diazo complex with fast blue RR indicator to give reddish coloured complex, indicating the activity of the phosphatase 43,44 (Figure 1 [45][46][47] . ...
Acid phosphatases (EC 3.1.3.2) are the enzymes that catalyse transphosphorylation reactions and promotes the hydrolysis of numerous orthophosphate esters in acidic media, as a crucial element for the metabolism of phosphate in tissues. Inorganic phosphate (Pi) utilisation and scavenging, as well as the turnover of Pi-rich sources found in plant vacuoles, are major processes in which intracellular and secretory acid phosphatases function. Therefore, a thorough understanding of these enzymes’ structural characteristics, specificity, and physiochemical properties is required to comprehend the function of acid phosphatases in plant energy metabolism. Furthermore, acid phosphatases are gaining increasing importance in industrial biotechnology due to their involvement in transphosphorylation processes and their ability to reduce phosphate levels in food products. Hence, this review aims to provide a comprehensive overview of the purification methods employed for isolating acid phosphatases from diverse plant sources, as well as their structural and functional properties. Additionally, the review explores the potential applications of these enzymes in various fields.
... The solution was filtered, and the filtrate was diluted tenfold by adding Tris-HCl buffer (pH 8.0). Phosphorus content was determined using the molybdenum blue method 25 . ...
Phosphorus is a key plant nutrient linked to plant growth during the early stages of primary succession in volcanic soils. Available phosphorus is thought to include soil and atmospheric phosphorus, but it is not well understood. Here, we focused on deposition as a potential phosphorus source. We evaluated the contribution of deposition to phosphorus uptake and growth in Fallopia japonica, a key pioneer species of primary succession. When we experimented with growing F. japonica under field conditions, F. japonica not covered by a roof absorbed more phosphorus than that covered by the roof, suggesting the influence of total (dry + wet) deposition. Furthermore, we tested the effects of deposition by treating F. japonica seedlings with wet deposition or distilled water in six volcanic soils. Plants that received the wet deposition treatment exhibited higher phosphorus contents and growth rates than those treated with distilled water. The phosphorus from wet deposition and the phosphorus from soil contributed nearly equally to F. japonica development. Our findings suggest that F. japonica grows during primary succession and builds up the phosphorus cycle by absorbing a trace amount of phosphorus from deposition and volcanic soils.
... SqwKL was assayed for its ability to catalyze ATP-driven ligation of sulfoacetate and CoA, which is the reverse of its proposed physiological reaction. Incubation of purified SqwKL with sulfoacetate, CoA, Mg 21 , and ATP led to the release of phosphate, as detected by a phosphomolybdate colorimetric assay ( Fig. 3A) (29). No reaction was observed in negative controls, in which either SqwKL or one of the three substrates was omitted. ...
Sulfoquinovose (SQ, 6-deoxy-6-sulfo-glucose) constitutes the polar head group of plant sulfolipids and is one of the most abundantly produced organosulfur compounds in nature. Degradation of SQ by bacterial communities contributes to sulfur recycling in many environments. Bacteria have evolved at least four mechanisms for glycolytic degradation of SQ, termed sulfoglycolysis, producing C3 sulfonate (dihydroxypropanesulfonate and sulfolactate) and C2 sulfonate (isethionate) by-products. These sulfonates are further degraded by other bacteria, leading to the mineralization of the sulfonate sulfur. The C2 sulfonate sulfoacetate is widespread in the environment and is also thought to be a product of sulfoglycolysis, although the mechanistic details are yet unknown. Here, we describe a gene cluster in an Acholeplasma sp., from a metagenome derived from deeply circulating subsurface aquifer fluids (GenBank accession no. QZKD01000037), encoding a variant of the recently discovered sulfoglycolytic transketolase (sulfo-TK) pathway that produces sulfoacetate instead of isethionate as a by-product. We report the biochemical characterization of a coenzyme A (CoA)-acylating sulfoacetaldehyde dehydrogenase (SqwD) and an ADP-forming sulfoacetate-CoA ligase (SqwKL), which collectively catalyze the oxidation of the transketolase product sulfoacetaldehyde into sulfoacetate, coupled with ATP formation. A bioinformatics study revealed the presence of this sulfo-TK variant in phylogenetically diverse bacteria, adding to the variety of mechanisms by which bacteria metabolize this ubiquitous sulfo-sugar. IMPORTANCE Many bacteria utilize environmentally widespread C2 sulfonate sulfoacetate as a sulfur source, and the disease-linked human gut sulfate- and sulfite-reducing bacteria can use it as a terminal electron receptor for anaerobic respiration generating toxic H2S. However, the mechanism of sulfoacetate formation is unknown, although it has been proposed that sulfoacetate originates from bacterial degradation of sulfoquinovose (SQ), the polar head group of sulfolipids present in all green plants. Here, we describe a variant of the recently discovered sulfoglycolytic transketolase (sulfo-TK) pathway. Unlike the regular sulfo-TK pathway that produces isethionate, our biochemical assays with recombinant proteins demonstrated that a CoA-acylating sulfoacetaldehyde dehydrogenase (SqwD) and an ADP-forming sulfoacetate-CoA ligase (SqwKL) in this variant pathway collectively catalyze the oxidation of the transketolase product sulfoacetaldehyde into sulfoacetate, coupled with ATP formation. A bioinformatics study revealed the presence of this sulfo-TK variant in phylogenetically diverse bacteria and interpreted the widespread existence of sulfoacetate.
... The optical absorbance of the solutions could be determined at ⊗ = 880 nm. 35 The adsorption efficiency Y (%) and capacity Q t (mg g −1 ) were calculated based on the following two formulas: ...
BACKGROUND
Zeolite NaP1 with high crystalline intensity and good shape has been synthesized by utilizing rice husk ash. Several factors related to the formation of zeolite, such as mixing ratios, aging time, and reaction time, were investigated. Furthermore, the potential phosphate adsorption of the optimal synthesized zeolite was evaluated.
RESULTS
Under the optimal conditions (SiO2/Al2O3 molar ratio = 4.0, NaOH concentration = 1.5 M, aging time = 12 h, and reaction time = 8 h), the surface area and pore diameter of the zeolite NaP1 obtained were 50 m² g⁻¹ and 40.77 Å, respectively. By varying the aging and reaction times, zeolite NaP1 with different surface areas could be created. Specifically, reducing the aging time and reaction time result in zeolite NaP1 with a lower surface area. The adsorption efficiency and adsorption capacity of the optimal synthesized zeolite NaP1 were 38.67% and 19.97 mg g⁻¹, respectively, at pH 6.0, with an initial phosphate concentration of 100 mg L⁻¹, an adsorbent dosage of 2 g L⁻¹, and a contact time of 120 min. The data are well described under the Temkin and Redlich‐Peterson models, with the correlation coefficients of 0.991 and 0.995, respectively. The maximum adsorption capacity of the Langmuir model reached 19.053 mg g⁻¹.
CONCLUSION
The main contribution of this work is the direct use of rice husk ash without pretreatment to generate high crystalline zeolite NaP1 in a shorter time. The phosphate ion removal efficiency by zeolite NaP1 may be further improved by functionalizing zeolite surfaces and expanding its other applications. © 2023 Society of Chemical Industry (SCI).
... The mixture was cooled and neutralized with 5 N NaOH solution in the presence of a universal indicator. Phosphorus content was determined by the phosphor-molybdenum method using ascorbic acid as a reducing agent [37]. The reaction mixture contained 0.4 mL of sample obtained after wet ashing, 2.6 mL of distilled water, 2 mL of solution containing 2.5% ammonium molybdate and 5 N H 2 SO 4 (1:1), and 1 mL of freshly prepared 1% ascorbic acid solution. ...
Plants’ mineral nutrition in acidic soils can be facilitated by phosphate solubilizing fungi inhabiting the root systems of these plants. We attempt to find dark septate endophyte (DSE) isolates in the roots of wild-heather plants, which are capable of improving plants’ phosphorus nutrition levels. Bright-field and confocal laser scanning microscopy were used for the visualization of endophytes. A model system of co-cultivation with Vaccinium macrocarpon Ait. was used to study a fungal isolate’s ability to supply plants with phosphorus. Fungal phytase activity and phosphorus content in plants were estimated spectrophotometrically. In V. vitis-idaea L. roots, we obtained a Phialocephala fortinii Wang, Wilcox DSE2 isolate with acid phytase activity (maximum 6.91 ± 0.17 U on 21st day of cultivation on potato-dextrose broth medium) and the ability to accumulate polyphosphates in hyphae cells. The ability of the isolate to increase both phosphorus accumulation and biomass in V. macrocarpon is also shown. The data obtained for the same isolate, as puzzle pieces put together, indicate the possible mediation of P. fortinii DSE2 isolate in the process of phosphorus intake from inorganic soil reserves to plants.
... Determination of LC 50 of potassium cyanide in Cat Fish (Clarias gariepinus) homogenate and the supernatant was used to determine the enzyme activity in treated cyanide fish tissues by determination of inorganic phosphate in the supernatant which is liberated during the hydrolysis of the substrate adenosine triphosphate at 37 o C. The method ofLowry and Lopez (1946) was used for inorganic phosphate assay. ...
The Present research studies the acute sub lethal toxicity of potassium cyanide (KCN) in the fresh water, Clarias gariepinus and the effects on its behaviour and the Na ⁺ - K ⁺ ATPase enzyme activity . Acute toxicity of potassium cyanide (free cyanide) to the freshwater fish was studied using static bioassay method over a period of 96 h. Different concentrations of the toxicant (KCN) were used and LC 50 value was found to be 361μg/L. Behavioural changes when exposed to lethal concentration of KCN showed increased feed intake, cannibalism, irregular swimming activity, rapid jerk movement, aggressiveness, loss of balance, opercula movement, surface behaviour, loss of equilibrium, change in body colour and convulsion. The effect of potassium on the Na ⁺ - K ⁺ ATPase of various physiological tissues which includes the gill, liver, muscles and intestinal over duration 0f 12 h-35 days were also studied. Cyanide intoxication resulted in marked changes in ATPase, shown by significant decrease in the enzyme activities. The result shows that ATPase enzyme together could be employed as a sensitive and useful biomarkers for cyanide pollution while the behavioural changes could be an early signal of toxicity.
... In Japan, a spectrophotometric molybdenum blue method for Pi determination is employed as the standard method [16], which is based on the method by Lowry and Lopez in 1946 [17]. By this method, Pi concentration can be determined between 0.1 and 3.0 mg/L Pi (3.2-96 μM) at a relative standard deviation (RSD) of 2-10%. ...
This chapter summarizes the developmental studies on environmental biosensors of enzymatic phosphate ion (Pi) biosensors for eutrophication and microbial biochemical oxygen demand (BOD) biosensors for organic pollution. In particular, an author focuses on the developmental studies that the author principally conducted, and describe the history and the insights into the future of these fields of environmental biosensors. In our developmental studies on the enzymatic Pi biosensors, we fabricated automatic instruments of a desktop-type and a submersible buoy-type, which was fabricated for remote biosensing of dam water. These instruments employed a luminol-chemiluminescence flow injection analysis (CL-FIA) system and enabled to have practical performances in precise Pi determination, operational stability, and accurate bioavailable Pi measurements. In the microbial BOD biosensor development, the author considered to apply the FIA concept enabling highly repeatable measurements to absorptiometric BOD measurements. Both precise temperature control and accurate time control to incubate measurement mixture of budding yeast cell suspension containing redox color indicator and sample enabled to obtain the highly repeatable results that led to highly sensitive BOD measurements. Looking back on our developmental studies, what the author was thinking at the time and the results obtained are described. Finally, the author discusses the developmental trends of these biosensor fields and new insights into the future perspectives.
... The digestion was carried out in a water bath at 80°C for 30 min. The concentration of Zn, Se, copper (Cu) and manganese (Mn) were measured in the digested samples using a novAA ® 400 PAtomic Absorption (Analytic Jena) by the method of Lowry and Lopez (1946). ...
... The concentration of minerals (Se and Zn) in samples were determined using a novAA ® 400 PAtomic Absorption (Analytic Jena, Germany) (Lowry & Lopez, 1946). In short, the frozen microalgae and rotifers were dried in an oven at 40°C for 24 h. ...
Rotifers are widely used in hatcheries to feed small‐sized aquatic larvae although one of their disadvantages is the lack of zinc and selenium 5‐ and 30‐fold lower than in copepods, respectively. To improve the rotifers quality, different concentrations of zinc and selenium (2, 4, 5 and 10 mg L−1 of each mineral) were added to the medium of the microalgae Isochrysis aff. galbana and Nannochloropsis oculata for 4 days, and then the microalgae were harvested and concentrated to feed the rotifers. N. oculata accumulated a greater amount of Zn and Se into cells than I. galbana. The cell size of algae given 0, 2, and 4 mg L−1 of minerals did not change in both microalgae, but enrichment of the microalgae with the 5 and 10 mg L−1 decreased the sizes and paled the colour of cells and increased cell division. The 2 mg L−1 was the best group for rotifers in terms of growth (population density, number of eggs, egg ratio, Specific growth rate, the maximum number and doubling time), and contained the second‐highest level of Zn (69.26 ± 0.60) and Se (103.5 ± 5.0) content within a safe limit. Thus, rotifers enriched with Se and Zn can be used as a mineral delivery method to cover the nutritional requirements of marine larvae.
... The Pi solution was filtered using a 0.22 mm PVDF syringe filter membrane prior to the colorimetric measurement. To these filtered solutions, ammonium molybdate-based reagent was added to produce a blue-colored solution which absorbance was measured at 880 nm wavelength [36]. ...
Background
The pressing demand to increase agricultural productivity amid the rapidly growing population has exponentially boosted fertilizers usage. Phosphate (Pi) runoff from fertilizers induces eutrophication in water sources and severely affects its surrounding ecosystems. To cope with Pi accumulation problem, this study reported the synthesis of an environmentally friendly magnetic adsorbent, namely Fe3O4/thiamine (thF).
Method
A one-step chemical oxidation and functionalization technique for thF synthesis was developed. X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), Fourier-transform infrared spectroscopy (FTIR), nitrogen (N2) sorption, and superconducting quantum interference device (SQUID) analysis were conducted to ensure the formation of Fe3O4, confirm the successful incorporation of thiamine, and gain insight into the factors influencing the adsorptivity of thF-363.
Significant Findings
The thF synthesized at 363 K (thF-363) produces an adsorbent with the highest Pi removal efficiency compared to other synthesis conditions. The thF-363 showed up to 1.51-fold higher adsorption capacity than the unmodified Fe3O4. The large surface area and occurrence of thiamine functional groups are the contributing factors in enhancing its adsorption capacity for Pi removal. The thF-363 did not adversely affect the growth of the model plant, Arabidopsis thaliana; demonstrating its suitability as an environmentally friendly adsorbents for Pi removal from eutrophicated water with the feasibility of magnetic separation from an aqueous system.
... The digestion was carried out in a water bath at 80°C for 30 min. The concentration of Zn, Se, copper (Cu) and manganese (Mn) were measured in the digested samples using a novAA ® 400 PAtomic Absorption (Analytic Jena) by the method of Lowry and Lopez (1946). ...
Rotifers are widely used as a live prey for finfish larvae, although their content in minerals such as selenium and zinc is lower than in fish. In the present study, mixed microalgae (i.e., Isochrysis aff. galbana and Nannochloropsis oculata) were enriched with different amounts of Se and Zn either alone (45, 90, and 135 mg L−1 of Zn, 60, 120, and 180 mg L−1 of Se) or mixed (22.5, 45, and 90 mg Zn L−1 and 30, 60, and 120 mg Se L−1). The content of minerals in the microalgae was analysed after 1 and 3 h enrichment and their effect on the growth and mineral composition of the rotifer. Based on the results, 1 h enrichment time and 90 and 120 mg L−1 of Zn and Se, respectively, resulted in the highest amount of Se and Zn in enriched microalgae. The maximum Zn content was detected in the rotifers fed 90 mg L−1 Zn and 90 mg L−1 Zn +120 mg L−1 of Se, but the maximum Se was observed in rotifers fed with 120 and 180 mg L−1 Se. Thus, Se‐ and Zn‐enriched rotifers can be used as a mineral delivery method to cover marine fish larvae nutritional requirements.
... Карбоангідразну активність вимірювали за допомогою скляного електрода за 2 °С в 14 мМ вероналовому буфері (рН 8,4), за наявності зразка починали реакцію додаванням води, насиченої СО 2 , за 0 °С та вимірювали зміни рН від 8,3 до 7,8; карбоангідразну активність подавали в мкмоль Н + на мг білка за хвилину з урахуванням буферної ємності середовища й зразків і визначали титруванням 0,1 М НС1 [14]; естеразну активність визначали за [15] та подавали в мкмоль п-нітрофенолу, що утворювався за хвилину після внесення субстрату, у розрахунку на 1 мг білка. АТФазну активність очищеного АТФ-синтазного комплексу визначали за кількістю утвореного фосфату, вимірюючи зміни його концентрації за методом Лоурі і Лопеса [16,17]. Сульфаніламадні інгібітори етоксизоламід (6-етокси-2-бензотіазолсульфонамід, Sigma) та ацетазоламід (N-[5-(аміносульфоніл)-1,3,4-тіадіазол-2-іл]ацетамід, Sigma) розчиняли в диметилсульфоксиді. ...
Aim. To isolate and purify protein complexes – ATP synthase and RuBisCO – from pea leaf chloroplasts and study the effect of a microbiological fertilizer “Extracon” and sulfonamide inhibitors acetazolamide and ethoxyzolamide on the enzymatic activity of these proteins.
Materials and methods. Chloroplasts were isolated from the leaves of two-week-old pea sprouts, protein complexes of purified thylakoid membranes were solubilized with digitonin (10 mg of digitonin per 1 mg of protein), the protein concentration was determined according to Lowry. Native electrophoresis with displacement of the charge of the soluble protein fraction from the chloroplast stroma, as well as membrane proteins, was carried out in the modified system of Anderson et al., Kolisnichenko et al. A modified Lemmley system was applied to the protein electrophoresis in the polyacrylamide gel in the presence of sodium dodecyl sulfate. The methods of Alain and Hintsik, as well as Gomorrah were used to determine the ATPase activity in the polyacrylamide gel. Visualization of the carbonic anhydrase activity in the polyacrylamide gel was performed by the method of Edwards and Petton.
Results and discussion. Using physicochemical methods of potentiometry, spectrophotometry the ATPase, carbonic anhydrase and esterase activities of the enzymes were studied. The results obtained indicate that specific carbonic anhydrase inhibitors (acetazolamide and ethoxyzolamide) also block the esterase and ATPase activity of the enzyme complexes. “Extracon” (a multifunctional microbiological preparation) almost 1.5 times increases the activity of the enzymes, showing a complex activating effect of the fertilizer on both light and dark reactions of photosynthesis.
Conclusions. The method of identification and isolation of RuBisCO and ATP synthase on the basis of two-dimensional electrophoresis and electrophoretic elution has been proposed. It allows determining the presence of certain enzyme activity of complexes at first in SDS plates (express analysis) and further to study the effect of various factors of endogenous and exogenous origin on the enzymatic properties of electrophoretically pure enzymes. The use of two-dimensional electrophoresis as a tool for assessing the impact of various factors of endogenous and exogenous origin on the plant cell and the plant as a whole through constant monitoring of the work and activity of enzyme systems of the plant cell is promising.
... Indeed, the values obtained by direct phosphate quantification were higher than those obtained from phytate dephosphorylation. This might be due to a dephosphorylation of other phosphorylated compounds present in the feed or feed ingredients (Lowry and Lopez, 1946) or an interaction between the phosphate released and other constituents of the feeds or feed ingredients. The initial IP2 and IP1 contents of feed ingredients was reported insignificant (Pointillart, 1994) and therefore only their IP3-IP6 contents were considered. ...
In vivo studies of the digestive process are long, expensive and difficult to rationalize, whereas in vitro systems may give more accessible insight into parts of this process. The purpose of this study was to show the ability of a three-step simulation of monogastric animals’ digestive system to estimate phytate hydrolysis and how it is affected by feed composition. Several feed ingredients: wheat, maize, soybean meal and rapeseed meal and complete diets: a wheat-maize-soybean-meal-based diet, a maize-soybean-meal diet and a wheat-maize-rapeseed-meal diet were treated using an adaptation of a described in vitro digestion simulation system in the presence of increasing doses of phytase. A strong dependence of phytate hydrolysis on the feed ingredient used was obtained: phosphorus releases were 0.3, 0.8, 1.0 and 1.6 g/kg at 0 U/kg of phytase supplementation for maize, soybean meal, wheat and rapeseed meal respectively and 1.2, 2.9, 1.7 and 3.9 at 1000 U/kg of bacterial phytase. The efficacy of enzymatic dephosphorylation of phytate was found dependent on the ingredient, which can be partially explained by their initial content in myo-inositol phosphates. The in vitro simulation was proven a useful tool to assess enzymatic dephosphorylation of phytate under different conditions.
... The isolates were analyzed for phosphate solubilization ability using tricalcium phosphate enriched Pikovskaya medium [7]. Phosphate solubilizing bacteria forming a clear halo zone around the colony were selected for their quantitative solubilization using ascorbic acid-molybdate Fig. 1 The practice of jhum cultivation in contrast to the change observed in recent trends (the steps involved in jhumming are given in uppercase letters) method [8]. Positive isolates were inoculated in Pikovskaya broth and grown in a shaker incubator at 160 rpm, 30°C for 7 days. ...
Bio-inoculants based on native Bacillus spp. have the potential to resuscitate the short fallow Jhum (slash and burn), degraded agro-ecosystem of Northeast India. Plant growth-promoting (PGP) native microbial isolates were investigated for reinvigorating growth of an upland rice variety (Bhalum). Of the 37 distinct bacterial isolates from fields left fallow for 2 years, 27% exhibited two or more PGP traits. Three native isolate strains SH2, MB1 and SS2 were selected as bio-inoculators, based on their levels of indole production, phosphate solubilization, siderophore and HCN production. They also showed no apparent toxicity on the tested rice seeds while enhancing the germination vigor of rice seeds with increased shoot length (39.7%), root length (37%) and plant height (32.7%) when compared to the untreated seeds. Under greenhouse conditions, plants derived from seeds treated with SH2, MB1 and SS2, displayed significant increments and enhanced growth biomass with 39.7% and 37.1% enhancement in shoot and root development versus the control. Phylogenetic analysis based on 16S rRNA sequences characterized SH2, MBI and SS2 as Bacillus luti, B. pacifus, and B. subtilis, respectively. The NCBI GenBank database and the phylogenetic dendrogram generated confirmed their identity with their related type strains. Their robust physiological response to temperature, pH and salt stress will bolster survival in the harsh slash-burn and shifting agriculture cycle. Consequently, application of SH2, MBI and SS2 would maximize growth productivity engendering the eco-restoration, sustainability and resolving low crop productivity of the short fallow Jhum agro-ecosystem
... The inhibitory concentration was determined by measuring the phosphorus content of the solution by the method of Lowry and Lopez [29]. N-methylated α,α-difluorophosphonates 9b and 10b show activity on E. coli DXR in the nanomolar concentration range and appear to be 2.5 to 5 times more efficient inhibitors than the parent compound fosmidomycin 3a (IC50 = 9 nM and 17 nM respectively vs. IC50 = 42 nM) and slightly less potent inhibitors than FR-900098 3b (IC50 = 4 nM). ...
Three α,α-difluorophosphonate derivatives of fosmidomycin were synthesized from diethyl 1,1-difluorobut-3-enylphosphonate and were evaluated on Escherichia coli. Two of them are among the best 1-deoxy-d-xylulose 5-phosphate reductoisomerase inhibitors, with IC50 in the nM range, much better than fosmidomycin, the reference compound. They also showed an enhanced antimicrobial activity against E. coli on Petri dishes in comparison with the corresponding phosphates and the non-fluorinated phosphonate.
... The reaction was incubated at 37°C in a water bath for 20 min and stopped by adding 1 ml of 10% trichloroacetic acid (TCA) and then centrifuged. The inorganic phosphorus (Pi) liberated was then estimated in the supernatant by the method of Lowry and Lopez (1946) which involved the use of ammonium molybdate and vitamin C. Assay for Na + /K + -ATPase (E.C.3.6.1.37) activity followed the procedure of Hesketh et al. (1978) as described by Afolabi et al (2016). ...
... The reaction was incubated at 37°C in a water bath for 20 min and stopped by adding 1 ml of 10% trichloroacetic acid (TCA) and then centrifuged. The inorganic phosphorus (Pi) liberated was then estimated in the supernatant by the method of Lowry and Lopez (1946) which involved the use of ammonium molybdate and vitamin C. Assay for Na + /K + -ATPase (E.C.3.6.1.37) activity followed the procedure of Hesketh et al. (1978) as described by Afolabi et al (2016). ...
... The amount of phosphorus in the sample solution was measured by the molybdenum blue colorimetric method 29 . ...
Plants have difficulty absorbing phosphorus from volcanic ash soils owing to the adsorption of phosphorus by aluminum and iron in the soils. Thus, on volcanic ash soils, the phosphorus source for natural vegetation is expected to be organic matter, however, there is a lack of experimental evidence regarding this occurrence. Here, we studied the effect of organic matter on plant growth of some species that occur in primary successions of volcanic ash soil ecosystems, based on growth experiments and chemical analyses. We found that a large amount of inorganic phosphorus (but only a limited amount of inorganic nitrogen) is leached from fresh leaf litter of the pioneer spices Fallopia japonica at the initial stage of litter decomposition. Phosphorus from the fresh litter specifically activated the growth of subsequently invading nitrogen-fixing alder when immature volcanic soil was used for cultivation. In contrast, old organic matter in mature soil was merely a minor source of phosphorus. These results suggest that fresh litter of F. japonica is essential for growth of nitrogen-fixing alder because the litter supplies phosphorus. We consider that rapid phosphorus cycles in fresh litter-plant systems underlie the productivity of natural vegetation even in mature ecosystems established on volcanic ash soils.
... Subsequent treatment with alkaline phosphatase ensures the release of the final phosphate from myo-inositol (IP1), which is relatively resistant to the reaction of phytase. The total phosphate released was measured using a colorimetric method (Fiske and Subbarow, 1925;Lowry and Lopez, 1946). For the analysis of minerals, samples of the diets, whole-body, muscle tissue and carapace were homogenised prior to analysis. ...
The presence of anti-nutritional factors such as phytic acid (PA) in a number of potentially useful plant ingredients has been a major impediment to their use as replacements for fishmeal in aquafeeds. This study builds on an earlier studying investigating the growth performance and nutrient utilisation in the juvenile Malaysian giant river prawn, Macrobrachium rosenbergii exploring the effects and interactions of dietary PA with minerals on growth (somatic tissue). Additionally, the study set out to determine the changes in whole-body mineral composition and also on muscle tissues and the carapace, when different levels of PA were included within the diet. The hypothesis under test was that PA, as a mineral-binding agent by chelating di- and trivalent cations inhibits mineral availability. To explore this, effects of graded levels of PA (i.e. 0.26–control, 6.48, 11.28, 16.53, 21.45 and 26.16 g PA kg⁻¹) on the moult frequency and mineral availability in juvenile M. rosenbergii (mean initial carapace length of 6.22 ± 0.52 mm; mean initial weight of 0.20 ± 0.01 g; n = 20 per replicate) fed over a period of 140 days were determined. The levels of PA assessed in this feed trial had no major detrimental effects on moult frequency. Negative effects (p < 0.05) of high PA levels (i.e. 21.45–26.16 g PA kg⁻¹), however, were found on the whole-body phosphorus concentration. An increasing trend, although not significantly so, was recorded for the calcium content in the whole-body and carapace with increasing PA inclusion. Significant changes (p < 0.05) were observed in the levels of zinc, copper, potassium and sodium in the carapace, particularly in the prawns fed the diet containing 11.28 g PA kg⁻¹, which suggests that the specific minerals were either selectively utilised or retained in the carapace. These changes may have a serious impairment hence caution should be exercised to ensure that the anti-nutritive effects of PA are minimised.
... Colorimetric determination of phosphorus was performed according to Fiske and Subbarow [18] and Lowry and Lopez [19]. One milliliter of the supernatant sample was added with 0.5 ml color reagent. ...
Soybean meal has long been considered as a plant protein source particularly for animal feedstuff. Nowadays, there is a promising prospect of this material for food processing. However, its protein utilization has drawbacks, such as poor protein solubility and anti-nutritional factors called phytic acid. The phytic acid could be reduced by the addition of phytase, whereas nutrient content enhanced by microbial fermentation. In this study, we analyzed the bio-modified of soybean meal (SBM) through fermentation by phytase producing Lactobacillus plantarum A1-E to evaluate its effect on nutritional quality and anti-nutrient factor called phytic acid and aflatoxin. The study was designated into two treatments consisted of unfermented soybean meal (USBM) and fermented soybean meal (FSBM) by L. plantarum A1-E. Parameters observed were microbial population, phytic acid content, aflatoxin content, and nutrient composition (moisture, ash, crude protein, crude fiber, and lipid). Data were analyzed using analysis of variance (ANOVA) and a T-test to compare treatment means. FSBM by L. plantarum A1-E increased population (6.8x10 ⁷ CFU/ml) during 72 h incubation (P<0.05) than 0 h (2.1x10 ⁶ CFU/ml). FSBM reduced phytic acid content (34.13%) and aflatoxin (6.12%) compared to control (USBM). Fermentation during 72 h had on average more crude protein (1.33-fold), crude fiber (2.09-fold), ash (1.13-fold), and less crude lipid (0.71-fold) than SBM. Thus, it can be concluded that FSBM by phytase producing L. plantarum A1-E could be applied for degradation phytic acid during food processing and represents an improvement in nutritional quality thereby becomes potential raw material as a new protein source for food.
... Chlorophyll (a, b and total chlorophyll content) and soluble protein concentration were estimated by the method of Arnon (1949) and that of Lowry et al. (1951), respectively. On the other hand, inorganic phosphate (Pi) by the method of Lowry and Lopez (1946) and protein phosphatase (Ppase) and acid phosphatase (AcPase) that of LeBel, Poirier and Beaudoin (1978) were determined. ...
p class="abstrakinggris">Waste released from sugar factories has a huge amount of inorganic and organic matter whose effect on plant is different when used in different concentrations. Hence, the effects of sugar factory effluent on the morphological and biochemical parameters of chickpeas ( Cicer arietinum Linn.) were studied by using the different concentrations of effluent for irrigation of chickpeas plant. The experiment was conducted at Botanical Garden, Department of Biochemistry, Bhaila (PG) College, Bhaila (Saharanpur), Uttar Pradesh. In the experiment, pots were arranged in a completely randomized design, replicated by six times and were labeled for the various treatments viz. 0%, 10%, 25%, 50%, 75% and 100% v/v. After that, chickpea plants were grown up to 10 days, in the soil irrigated with different concentrations of sugar factory effluent. It was observed that sugar factory effluent promoted the growth of chickpea seeds at 10–25% concentration, but its lethal impacts were noted when concentrations were above 50–100%. Hence, it was found that at lower concentrations (upto 25%), sugar factory effluent behaves as compost for better growth of chickpeas, but at high concentrations, it shows toxicity and behaves as a pollutant. Moreover, it makes the speed of biochemical reactions slow in chickpeas. Therefore, sugar factory effluent can be used for irrigation of chickpeas after proper dilution . </p
... The Pi containing solution will produce a colored solution with maximum absorbance at 880 nm. Details of the measurement procedure can be found elsewhere (Lowry and Lopez, 1946). ...
Organo-bentonite (OrB) was prepared by modifying bentonite with chitosan, and natural surfactant extracted from Sapindus rarak fruit. The physical alteration post-modification, performance of phosphates (Pi) adsorption, and possibility as a Pi-supplementation for plants of OrB were assessed and compared to acid-activated bentonite (AAB). The physical alteration due to modification of bentonite was characterized. SEM images were not indicating significant morphology differences between OrB and AAB. Existence of chitosan layers in OrB causes a decrease in basal spacing as characterized using XRD. The BET surface area of OrB was decreased compared to AAB due to pore coverage by chitosan. Adsorption studies reveal that OrB has a higher adsorption capacity towards Pi than AAB, which is 97.608 and 131.685 mg/g at 323 K for AAB and OrB, respectively. The H-shape isotherm curve indicates that chemisorption is dominantly controlling the adsorption. The isotherm and kinetics adsorption were well fitted to Langmuir and Pseudo-second order models, respectively. Performance of AAB and OrB as Pi-supplementation was assessed based on growth phenotypes of Arabidopsis thaliana; seedlings show that supplementation of [email protected] and [email protected] (at half doses) can promote primary root extension. These results also demonstrate the safety of direct disposal of the materials into the soil.
... The P i content in the water sample was determined within one week after collection. P i assay was performed according to the method of Lowry and Lopez (1946) with slight modification. The reaction mixture consisted of 2 ml of deionized water (blank) or sample, 0.2 ml of 1% ascorbic acid and 0.2 ml of 1% ammonium molybdate in 0.025 mol l -1 H 2 SO 4 . ...
Giant clams live in symbiosis with extracellular dinoflagellates (zooxanthellae) in oligotrophic tropical waters of the Indo-Pacific. Zooxanthellae are found mainly in the colourful outer mantle. They donate photosynthates to the host during insolation, while the host absorbs and supplies them with exogenous nutrients, including inorganic phosphate (Pi). We report for the first time that the fluted giant clam, Tridacna squamosa, could absorb Pi at a higher rate in light than in darkness. We had also obtained the complete coding cDNA sequence of a homolog of sodium-dependent phosphate transport protein 2a (NPT2a-like) from its ctenidium, which comprised 1740 bp, coding for 580 amino acids of 63.8 kDa. The gene, NPT2a-like, was expressed strongly in the colorful outer mantle, the whitish inner mantle and the ctenidium, but weakly in all other organs examined including the kidney. The protein, NPT2a-like, was localized apically in the epithelium covering the ctenidial filaments, the upper epithelium of the outer mantle and the seawater-facing epithelium of the inner mantle. As these epithelia are in contact with seawater, all three organs could probably absorb Pi through the apical NPT2a-like. Nonetheless, the outer mantle could be a major site of light-enhanced Pi absorption, as the protein abundance of its NPT2a-like, but not those in the other two organs, was up-regulated by illumination. Photosynthesizing zooxanthellae need Pi, and the outer mantle could deliver the absorbed Pi directly to the zooxanthellae therein, reducing their need to compete with other host organs for the Pi absorbed by the ctenidium and circulating in the hemolymph.
... The pH of the resulting solution was adjusted to the desired value by 1 M NaOH and precipitation was detected by visual inspection. After 24-h incubation at room temperature, the mixtures were centrifuged and the amount of P i in supernatants was determined according to Ref. [92]. Briefly, 20 µL of supernatant was withdrawn and diluted with 0.78 mL of water. ...
To investigate divalent metal ion (Me²⁺) requirements in electrophilic biocatalysis, we compared Mg²⁺, Mn²⁺, Co²⁺, Zn²⁺, Cu²⁺, Ni²⁺, Cd²⁺, Ca²⁺, and Fe²⁺ activities with 13 enzymes executing nucleotidyl and/or phosphoryl transfer. We find that each Me²⁺ ion was highly catalytically active with one or more of the related enzymes. This result suggests that features of Me²⁺ coordination at the active center, and/or the enzyme‐mediated presentation of the reactants to the chelated Me²⁺, rather than the nature of the Me²⁺, determine the ability of the Me²⁺ to support catalysis. At physiological pH, all the tested Me²⁺ ions, with the exception of Mg²⁺, produced insoluble complexes with inorganic phosphate (Pi) and bicarbonate ( HCO3-). These data suggest that early in the development of life, bioavailability and biocompatibility with these abundant cellular metabolites may have been decisive factors in the choice of Mg²⁺ as the major ion for biocatalysis. Taking into account the concentrations of inorganic ions in the ancient environment in which the first cells emerged, as inferred from the ‘chemistry conservation principle’, the choice of Mg²⁺ was predetermined prior to the origin of life.
... Substrate specificity test for alkaline phosphatase was done by analyzing inorganic phosphate obtained from hydrolysis of monosubstituted phosphate linkages compounds by alkaline phosphatase. Lowry-Lopez method (23) was used to determine the concentration of released inorganic phosphate. The reaction mixture containing alkaline phosphatase enzyme and phosphorylated compounds (5.4 mM-Tris-HCl, pH 9.5) was incubated at 50 ˚C for 20 min. ...
Some soil microbes have the capability to solubilize mineral phosphate into organic phosphorous and used as biofertilizer to improve crop productivity in agricultural field. In this study, phosphate solubilization assay was carried out onto media plates containing calcium phsophate precipitated nutrient agar media for bacterial strains like Bacillus megaterium MTCC 453, Bacillus subtilis MTCC 1134, Bacillus licheniformis MTCC 2312, Pseudomonas aeruginosa MTCC 424, Escherichia coli MTCC 570. Among these bacterial strains, B. licheniformis MTCC 2312 showed largest clear zone of phosphate solubilzation and maximum activity of alkaline phosphatase. The enzyme alkaline phosphatase was purified from B. licheniformis MTCC 2312 with purification fold 3.52 and specific activity 295.89 Unit/mg protein using DEAE-sepharose chromatography. This enzyme showed molecular weight as 60 KD, thermostability upto 50˚C, pH stability up to 8.5 and Michaelis constant (Km) and maximum activity (Vmax) as 2.30 mM and 2223 U/ml respectively. The lyophilized powder of this enzyme was further supplemented with media components for the growth of Zea mays for carrying tissue culture experiment. The sterilized soil supplemented with alkaline phosphatase improved the total height, dry weight, % phosphate content in the stem and root of Zea mays by 3.07, 3.15, 2.35 and 1.76 fold respectively compared to control set. This enzyme could be used at large extent as effective biofertilizer for the agricultural industry.
Intestinal transference of calcium and rate of bone turnover were evaluated in ovariectomized rats fed for 15
days with a high amount (30%) of lipid enriched with monounsaturated (groundnut oil), polyunsaturated
(sunflower oil) and saturated (coconut oil) fatty acids. The results were compared with those for sham-operated
control and ovariectomized groups fed a normal diet (7% groundnut oil). Irrespective of the saturation and
unsaturation characteristics, all lipids (edible oils) used in our study considerably decreased the rate of in situ
intestinal transference of calcium. Likewise, the activities of intestinal mucosal enzymes, alkaline phosphatase
(AP) and calcium ATPase (Ca2+-ATPase) were decreased significantly in all the segments of the small intestine
in a descending gradient. Significant changes in bone turnover and bone calcium (Ca) mobilization were
confirmed in these animals by marked alterations in plasma AP activity, urinary calcium and phosphate
excretion and calcium to creatinine (Ca:creatinine) ratio. Lipid supplementation (30%) in such ovariectomized
rats using groundnut oil (monounsaturated), sunflower oil (polyunsaturated) or coconut oil (saturated) for 15
days further enhanced all of the above observed parameters. These results suggest that the intake of high
amounts of lipids with different unsaturation and saturation characteristics may be an important factor in
determining bone loss in ovariectomized rats.
Key words: ovariectomy, high lipid diet, oils, saturated, unsaturated, intestinal, epithelium, calcium, bone turnover,
osteoporosis
Despite the positive role of rotifers in many hatcheries for feeding the early stages of fish larvae and crustaceans, the lower content of minerals such as selenium (Se, 30 times) and zinc (Zn, 5 times) is considered as one of the disadvantages of this live prey compared to copepods. Therefore, increasing the amount of these nutrients through enrichment is essential. To this end, the present study investigated the effects of the combination of 2 algae Isochrysis aff. galbana and Nannochloropsis oculata enriched with sodium selenite and zinc sulfate on growth and mineral content of rotifer. First, the 1:1 composition of the algae was enriched with a mixture of zinc and selenium salts (each with concentrations of 20, 40 and 80 mg L-1 in algal enrichment medium). The highest number of rotifers (339 ± 11.06 ind mL-1), SGR (± 0.48 ± 0.008 day-1), and the lowest doubling time without any significant difference observed in rotifers fed with algae enriched with 40 mg L-1 of both minerals (p>0.05). However, the highest amount of Zn and Se were observed in those fed with 80 mg L-1 of both minerals. In conclusion, feeding rotifers with Se and Zn enriched algae increased Se and Zn in the rotifers. Thus, microalgae enriched with Se and Zn can be used to feed marine rotifer to meet the nutritional requirements.
Glycoside phosphorylases (GPs), which catalyze the reversible phosphorolysis of glycosides, are promising enzymes for the efficient production of glycosides. Various GPs with new catalytic activities are discovered from uncharacterized proteins phylogenetically distant from known enzymes in the past decade. In this study, we characterized Paenibacillus borealis PBOR_28850 protein, belonging to glycoside hydrolase family 94. Screening of acceptor substrates for reverse phosphorolysis, in which α- d -glucose 1-phosphate was used as the donor substrate, revealed that the recombinant PBOR_28850 produced in Escherichia coli specifically utilized d -galactose as an acceptor and produced solabiose (β- d -Glc p -(1 → 3)- d -Gal). This indicates that PBOR_28850 is a new GP, solabiose phosphorylase. PBOR_28850 catalyzed the phosphorolysis and synthesis of solabiose through a sequential bi-bi mechanism involving the formation of a ternary complex. The production of solabiose from lactose and sucrose has been established. Lactose was hydrolyzed to d -galactose and d -glucose by β-galactosidase. Phosphorolysis of sucrose and synthesis of solabiose were then coupled by adding sucrose, sucrose phosphorylase, and PBOR_28850 to the reaction mixture. Using 210 mmol lactose and 280 mmol sucrose, 207 mmol of solabiose was produced. Yeast treatment degraded the remaining monosaccharides and sucrose without reducing solabiose. Solabiose with a purity of 93.7% was obtained without any chromatographic procedures.
Phaseolus vulgaris L. (Fabaceae) is one of the most important crops and one of the main protein sources. The objective of this research was to evaluate the effect of the inoculation of Rhizophagus irregularis and Rhizobium sp. on the growth and production of P. vulgaris. Plantlets were inoculated with both microorganisms and we measured the number of leaves, knots, stem diameter, shoot length, leaf area, fresh and dry weight, total protein content and yield components. Rhizobium sp. promoted plant growth, increasing the shoot diameter, dry weight, leaf area, total protein content and yield. Mycorrhizal fungi did not affect the number of knots, leaves, shoot length, protein content or fresh and dry weight. However, differences were observed in the yield components number and weight of seeds. Coinoculation with both microorganisms promoted an increase in the growth parameters (number of leaves, leaf area, fresh and dry weight, shoot length and stem diameter) and at the same time caused an increase in yield components (seed number and seed weight). It is concluded that the inoculation of both Rhizophagus irregularis and Rhizobium sp., promotes the growth, development and yield in Phaseolus vulgaris var. Canario "canary bean" under greenhouse conditions.
Glycoside phosphorylases (GPs) are a major group of enzymes which catalyze the reversible formation and cleavage of glycosidic linkages, thus combining a dual functionality of both glycoside hydrolases (GHs) and glycosyltransferases (GTs). This unique advantage of reversibility, alongside with the strict regioselectivity, and the use of simple sugar phosphate donors, makes phosphorylases valuable catalysts in the synthesis of various carbohydrate structures, such as the synthesis of low-calorie sweeteners and valuable disaccharides, the synthesis of prebiotic compounds linked to health benefits, synthetic drug delivery systems and other physiological related structures, including, core components of N-linked glycans, O-linked mucin glycoproteins. Although these types of enzymes are still under-explored compared to the glycoside hydrolases and the glycosyltransferases, recently more discovered phosphorylases had been reported leading to more possibilities of synthesizing difficult glycosides. Herein we provide an overview of the distribution of GPs within the carbohydrate-active enzymes (CAZy) database, and the recent utilization of these enzymes for synthetic approaches of di- and oligosaccharides.
A close relationship between angiotensin II (ANG II) and the renal dopaminergic system (RDS) has been reported. Our aim was to study whether renal dopamine and ANG II can interact to modify renal sodium handling and then to elucidate the related mechanism. Anesthetized male Sprague-Dawley rats were used in experiments. ANG II, exogenous dopamine, and decynium-22 (or D-22, an isocyanine that specifically blocks electrogenic organic cation transporters, OCTs), were infused in vivo for 120 min. We analyzed renal and hemodynamic parameters, renal Na+, K+-ATPase levels, OCT activity, and urinary dopamine concentrations. We also evaluated the expression of D1 receptor, electroneutral organic cation transporters (OCTNs), and OCTs. ANG II decreased renal excretion of sodium in the presence of exogenous dopamine, increased Na+, K+-ATPase activity, and decreased the urinary dopamine concentration. D-22 treatment exacerbated the ANG II-mediated decrease in renal excretion of sodium and dopamine urine excretion but did not modify ANG II stimulation of Na+, K+-ATPase activity. The infusion of ANG II did not affect the expression of D1 receptor, OCTs, or OCTNs. However, the activity of OCTs was diminished by the presence of ANG II. Although ANG II did not alter the expression of D1 receptor, OCTs, and OCTNs in renal tissues, it modified the activity of OCTs and thereby decreased the urinary dopamine concentration, showing a novel mechanism by which ANG II decreases dopamine transport and its availability in the tubular lumen to stimulate D1 receptor. This study demonstrates a relationship between ANG II and dopamine, where both agents counteract their effects on sodium excretion.
In this study, feeding experiment and subsequent digestibility trial were performed to investigate the utilization of extruded soybean meal (SBM) and corn gluten meal (CGM) as feed ingredients for juvenile rainbow trout. Plant ingredients have undergone extrusion at low temperature (100°C, LT) or high temperature (150°C, HT) for 30 s. Four isonitrogenous (44%, crude protein) and isolipidic (14%, crude lipid) diets were formulated. Control diet is fishmeal based while a combined (1:1) non‐extruded SBM and CGM for NE diet, LT SBM and LT CGM for LT diet and HT SBM and HT CGM for HT diet. Two hundred forty rainbow trout juveniles (7.8 g average body weight) were randomly divided into 12 rectangular 60 L glass aquaria and offered four different diets in triplicate. Fish were fed at satiation twice a day, six days a week for 12 weeks. Phosphorus in phytic acid level of extruded ingredients decreased through extrusion cooking. Final weight, weight gain, SGR and PER of fish fed HT diet were significantly (p < .05) higher than those fed with NE diet. Apparent digestibility coefficient for protein of LT and HT diets is significantly higher than of NE diet. The results of this study demonstrated that HT extruded SBM and CGM are suitable feed ingredients for rainbow trout diet without compromising fish growth, feed utilization and fish body composition.
The efficiency of electron carriers in thylakoid membranes untreated and exposed to heat 7-day-old barley seedlings was evaluated with PAM fluorescence. Darkness–light transitional states in chloroplasts after heat exposure are studied. Thermoinduced changes in linear and cyclic electron transport chain of chloroplasts are revealed. The activation of NADPH-dependent electron flux after exposure to elevated temperatures is shown. We assumed that ΔрН of thylakoid membranes employed the regulatory role in the distribution of electron flows and the adaptation of the photosynthetic apparatus to stressful effects.
Schizophrenia is a devastating illness and displays a wide range of psychotic symptoms. Accumulating evidence indicate impairment of bioenergetic pathways including energy storage and usage in the pathogenesis of schizophrenia. Although well-established synthetic drugs are being used for the management of schizophrenia, most of them have several adverse effects. Hence, natural products derived from medicinal plants represent a continuous major source for ethnomedicine-derived pharmaceuticals for different neurological disorders including schizophrenia. In the present study, we have investigated the neuroprotective effect of the novel bioactive compound i.e. “3-(3,4-dimethoxy phenyl) -1- (4-methoxyphenyl) prop-2-en-1-one” of Celastrus paniculata against ketamine-induced schizophrenia with particular reference to the activities of ATPase using in vivo and in silico methods. Ketamine-induced schizophrenia caused significant reduction in the activities of all three ATPases (Na⁺/K⁺, Ca²⁺ and Mg²⁺) in different regions of brain which reflects the decreased turnover of ATP, presumably due to the inhibition of oxidoreductase system and uncoupling of the same from the electron transport system. On par with the reference compound, clozapine, the activity levels of all three ATPases were restored to normal after pretreatment with the compound suggesting recovery of energy loss that was occurred during ketamine-induced schizophrenia. Besides, the compound has shown strong interaction and exhibited highest binding energies against all the three ATPases with a lowest inhibition constant value than the clozapine. The results of the present study clearly imply that the compound exhibit significant neuroprotective and antischizophrenic effect by modulating bioenergietic pathways that were altered during induced schizophrenia.
Previously we have discovered that titanium (Ti) binds with bone phosphoproteins SIBLING protein family, by using a Ti beads chromatography. Furthermore, we showed that the isolated bone phosphoproteins remarkably enhanced bone formation when we coated the Ti device with them and implanted into rat calvaria. Therefore, we have called the Ti-binding bone phosphoproteins as “the implant proteins.” This discovery encouraged us to create a new biomolecule that can simulate the functions of the implant proteins. Since significant characteristics of the implant proteins are the presence of multiple phosphate groups and the occurrence of single cell-adhering RGD sequence, we decided for the first place to phosphorylate chitin and collagen to see whether they acquire or increase Ti-binding ability. Results showed that more than 70% of phosphorylated chitin bound with Ti, and phosphorylated collagen enhanced about 7% of its Ti-binding ability. These modified biomolecules, P-chitin and P-collagen will become highly useful for new development of Ti-related bone regenerative medicine.
Trehalose 6-phosphate (Tre6P) is an important intermediate for trehalose biosynthesis. Recent researches have revealed that Tre6P is an endogenous signaling molecule that regulates plant development and stress responses. The necessity of Tre6P in physiological studies is expected to be increasing. To achieve the cost-effective production of Tre6P, a novel approach is required. In this study, we utilized trehalose 6-phosphate phosphorylase (TrePP) from Lactococcus lactis to produce Tre6P. In the reverse phosphorolysis by the TrePP, 91.9 mM Tre6P was produced from 100 mM β-glucose 1-phosphate (β-Glc1P) and 100 mM glucose 6-phosphate (Glc6P). The one-pot reaction of TrePP and maltose phosphorylase (MP) enabled production of 65 mM Tre6P from 100 mM maltose, 100 mM Glc6P, and 20 mM inorganic phosphate. Addition of β-phosphoglucomutase to this reaction produced Glc6P from β-Glc1P and thus reduced requirement of Glc6P as a starting material. Within the range of 20-469 mM inorganic phosphate tested, the 54 mM concentration yielded the highest amount of Tre6P (33 mM). Addition of yeast increased the yield because of its glucose consumption. Finally, from 100 mmol maltose and 60 mmol inorganic phosphate, we successfully achieved production of 37.5 mmol Tre6P in a one-pot reaction (100 mL), and 9.4 g Tre6P dipotassium salt was obtained.
- H M Kalckar
Kalckar, H. M., J. Biol. Chem., 168,723 (1945).
- F Lipmann
- L C Tuttle
Lipmann, F., and Tuttle, L. C., J. Biol. Chem., 163,571 (1944).
- C H Fiske
- Y Subbarow
Fiske, C. H., and Subbarow, Y., J. Biol. Chem., 66, 375 (1925).
- C H Fiske
- Y Subbarow
Fiske, C. H., and Subbarow, Y., J. Biol. Chem., 81,629 (1929).
- R Ammon
- K Hinsberg
Ammon, R., and Hinsberg, K., Z. physiol.
Chem., 239,207
(1936).