The study was designed to purify enzyme from weed plant in association with wheat crop to assess and compare the purification and biochemical properties with acid phosphatases of seedlings of various ...plants. Acid phosphatase from seedlings of Rumex dentatus (Curly dock) was purified by different chromatography and chromatofocusing techniques with specific activity of 63U/mg of protein. The yield was about 3%. Single band was detected on SDS-polyacrylamide gel electrophoresis confirming the enzyme was homogeneous. Apparent molecular weight of 48 kDa was obtained. Gel filtration experiment indicated that native enzyme had a approximately molecular weight of 96 kDa, suggesting that this enzyme is homodimer. The enzyme showed Km value of 0.5 mM and Vmax of 60 and#181;M of p-nitrophenyl phosphate hydrolysed/min/mg of protein under experimental conditions. The optimum pH of 5.0 and temperature of 50and#176;C were obtained. Divalent cations such as copper and zinc ions caused acid phosphatase inhibition but the presence of 20 mM EDTA in the enzyme-metal ions incubation mixtures reversed the enzyme inhibition to some extent. The activity of 54 % and 63% were recovered back, respectively. Ca2+ and Mg2+ had very small activating effect on activity in the absence or presence of EDTA. The reaction of enzyme with iodoacetic acid or N-ethyl-maleimide had no inhibitory effect, pointing to a non-involvement of cysteine residues in enzyme action. Further, β-mercaptoethanol or dithiothreitol at low concentrations had very little activating effect revealing that SH-group containing amino acid in the enzyme may not be significant for its catalytic activity. The pH dependent variation of Km study showed that histidine may constitute a part of the active site. Acid phosphatase was competitively inhibited by phosphate and vanadate. Fluoride and Zn2+ acted as non-competitive inhibitors while molybdate showed mixed type of inhibition.
Inherent biases in numerical simulation models of global climate models (GCMs) reduce the scope for accurately assessing future droughts under multimodel ensemble. The aim of this research is to ...increase the efficiency of multimodel ensembles. Consequently, this paper introduces a two-way hybrid weighting scheme for ensembling multiple GCMs. The proposed weighting scheme enhances the coherence of the multimodel ensemble of climate model simulations with real observed data and minimizes the impact of non-identical seasonal behavior in climate simulations within the multimodel ensemble. In the application, we consider precipitation data from 18 GCMs of CMIP6 from the Tibet Plateau region. To evaluate the effectiveness of the proposed scheme, we compared the proposed ensemble data with simple model averaged (SMA) ensemble using the Pearson product-moment correlation coefficient (
r
), root mean squared error (RMSE), and mean absolute error (MAE). Furthermore, we assessed the future characteristics of various drought categories using the steady-state probabilities of a Markov Chain. Small values of errors metircs and high correlation indicate that the proposed hybrid weighting scheme exhibits better performance than the SMA. Under the proposed index, the long term probabilities of various drought classe show variations across different time scales of drought. These results provide perspectives on how the the long term probabilities of various drought categories evolve over time.
Enterobacter cloacae is mainly responsible for sepsis, urethritis, and respiratory tract infections. These bacteria may affect the transcription of the host and particularly their immune system by ...producing changes in their epigenetics. In the present study, four proteins of Enterobacter cloacae were used to predict the epitopes for the construction of an mRNA vaccine against Enterobacter cloacae infections. In order to generate cellular and humoral responses, various immunoinformatic-based approaches were used for developing the vaccine. The molecular docking analysis was performed for predicting the interaction among the chosen epitopes and corresponding MHC alleles. The vaccine was developed by combining epitopes (thirty-three total), which include the adjuvant Toll-like receptor-4 (TLR4). The constructed vaccine was analyzed and predicted to cover 99.2% of the global population. Additionally, in silico immunological modeling of the vaccination was also carried out. When it enters the cytoplasm of the human (host), the codon is optimized to generate the translated mRNA efficiently. Moreover, the peptide structures were analyzed and docked with TLR-3 and TLR-4. A dynamic simulation predicted the stability of the binding complex. The assumed construct was considered to be a potential candidate for a vaccine against Enterobacter cloacae infections. Hence, the proposed construct is suitable for in vitro analyses to validate its effectiveness.
Water purification is one of the worldwide problem and most of the conventional methods are associated with a number of drawbacks. Therefore it is the need of the day to develop new methods and ...materials to overcome the problem of water purification. In this research work we present a simple and green approach to synthesize silver decorated titanium dioxide (Ag/TiO2) nanocomposite with an efficient photo catalytic activities. Phytochemicals of the Cestrum nocturnum leaf extract were used to synthesize silver nanoparticles (AgNPs), Titanium dioxide (TiO2) and Ag/TiO2 nanocomposite. To confirm the formation, crystal structure, particle size and shape of green synthesized nanoparticles and nanocomposite, they were characterized by UV–visible spectroscopy (UV–vis), X-ray diffraction spectroscopy (XRD), high resolution transmission electron microscopy (HRTEM), Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FT-IR). The AgNPs, TiO2 and Ag/TiO2 were evaluated for photo degradation of methylene blue (MB) and photo inhibition of Bacteria. The bio-synthesized Ag/TiO2 nanocomposite was observed to have strong catalytic activities for photo reduction of MB and photo inactivation of bacteria as compared to bare AgNPs and TiO2. In the presence of Ag/TiO2, 90% of MB was degraded only in 40min of irradiation. Alternatively the bare AgNPs and TiO2 degraded less than 30% and 80% respectively of MB even in more than 100min of irradiation. Similarly the Ag/TiO2 has very strong photo inhibition efficiency towards Escherichia coli and Pseudomonas aeruginosa. The zone of inhibition of irradiated Ag/TiO2 nanocomposites against E. coli and P. aeruginosa was 19mm and 17mm respectively which was two times higher than in dark. These promising photocatalytic activities of nanocomposite may be due to the highly decorated AgNPs over the surface of TiO2.
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•Preparation of Ag/TiO2 by a novel green method•Cestrum nocturnum leaf extract was used as a reducing source•Photo inhibition activity of TiO2 and Ag/TiO2 against two bacteria•MIC, durability and effect of irradiation time was also analyzed•Photo degradation of methylene blue by Ag/TiO2 and TiO2
Low molecular weight acid phosphatase (LM-ACP) peak 2 (the isoenzyme corresponding to isoform 2, IF-2) from the liver of fish Rahu (Labeo rohita) was purified to homogeneity. 900 times purification ...resulted with specific activity of 35 U/mg of protein and recovery of 0.2%. The enzyme was found homogeneous on sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Molecular weight of 18 killo Daltons (kDa) was obtained. The peak 1 isoenzyme corresponding to isoform1 (IF-1) was partially purified about 160 times with specific activity of 7 U/mg of protein. Major protein band corresponding to 18 kDa was seen along with other protein faint bands. High molecular weight acid phosphatase (HM-ACP) was also partially purified. The molecular weight was estimated to be a 100 kDa by gel filtration on Sephadex G-100. LM-ACP isoenzymes and HM-ACP enzyme were studied for their substrate specificity, sensitivity to inhibitors or activators and other kinetic parameters. LM-ACP isoenzymes were not inhibited by tartrate and fluoride but were inhibited by sulfhydryl reagent whereas high molecular weight enzyme was strongly inhibited by fluoride and tartrate. Phosphate vanadate and molybdate inhibited both types of enzymes competitively, but their action was more pronounced in HM-ACP enzyme. LM-ACP was effectively activated by purine compounds whereas HM-ACP was not. LM-ACP showed strict substrate specificity while HM-ACP showed broad substrate specificity. The two types of acid phosphatases also differed in their rate of hydrolysis of α-naphthyl phosphate and β-glyerophosphate.
This study was conducted to investigate the factors affecting the adoption of organic farming in Peshawar-Pakistan. A total of 100 respondents were randomly selected from the four different ...cultivated areas of Peshawar, namely Palosi, Regi, Ternab and Pushtakhara. Binary logistic regression was used in this study to categorize the organic farming into adoption and non-adoption. The purpose of this model was to check the event probability for a categorical response variable with two outcomes. The results of the binary logistic show that factors affecting adoption of organic farming have a significant effect on the farmer productivity. Moreover, cost, productivity, profitability, compatibility and efficiency have a positive and significant effect. Thus, it is obvious that adopting organic farming not only to increase the farmer income but also to protect environmental pollution by avoiding the toxic chemical and fertilizer. Finally, we suggest that government agencies, extension and research institution should play a vital role to strengthen the awareness and advantages of organic farming.
Herein for the first time a novel acid phosphatase from the seedlings of Cichorium intybus was purified to homogeneity by using various chromatographic techniques (salt precipitation, ion exchange, ...size exclusion and affinity chromatography) and thermodynamically characterized. The molecular mass of purified enzyme (66 kDa) was determined by SDS-PAGE under denaturing and non-denaturing conditions and by gel-filtration confirmed as dimer of molecular mass 130 kDa. The Michaelis-Menten (Km) constant for -p-NPP (0.3 mM) and (7.6 μmol/min/mg) Vmax. The enzyme was competitively inhibited by phosphate, molybdate and vanadate. Phenyl phosphate, ɑ and β-glycero-phosphate and-p-NPP were found to be good substrate. When temperature increased from (55 °C to 75 °C), the deactivation rate constant (kd) was increased (0.1 to 4.6 min−1) and half- life was decreased from 630 min to 15 min. Various thermal denaturation parameters; change in enthalpy (ΔH°), change in entropy (ΔS°) and change in free energy (ΔG°) were found 121.93 KJ·mol−1, 72.45 KJ·mol−1 and 98.08 KJ·mol−1 respectively, confirming that acid phosphatase undergoes a significant process of unfolding during deactivation. The biochemical properties of acid phosphatase from C. intybus on the behalf of biological activity and its relationship to pH variations, thermal deactivation and kinetics parameters provide an insight into its novel features.
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Herein acid phosphatase isoenzyme was extracted from the C. murale seedlings. The purification was accomplished by chromatographic techniques and passing through DEAE-cellulose and Sephadex G-100 ...column. The specific activity of acid phosphatase 5.75 U/mg of protein was obtained with 66 purification fold 15.8% yield and molecular mass was 29 kDa with very faint bands corresponding to 18 kDa and 14 kDa. The maximal activity at pH 5.0 and 50 °C best illustrated by first order kinetics. When temperature was raised (55 °C to 75 °C), the deactivation rate constant was increased from 0.001 to 0.014 min−1, while half-life was decreased from 693 to 49 min−1. The results of activity collected at different temperature were then used to estimate, activation energy of hydrolysis reaction (Ea = 47.59 kJmol−1). A high Z-value (18.86 °C min−1) was obtained indicating a less sensitivity towards temperatures. The residual activity examinations were carried out from 55 °C to 75 °C and assessing the Deactivation Energy (Ed 116.39 kJmol−1), Enthalpy change (ΔH° 113.55kJmol−1), Entropy change (ΔS° 110.33kJmol−1) and change in Gibbs free energy (ΔG° 10.02 kJmol−1). Taken together, thermodynamic parameters confirm the high stability of enzyme and show potential commercial applicability.
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A low molecular weight acid phosphatase was purified to homogeneity from chicken heart with a specific activity of 42 U/mg and a recovery of about 1%. Nearly 800 fold purification was achieved. The ...molecular weight was estimated to be 18 kDa by SDS-polyacrylamide gel electrophoresis. Para-nitrophenyl phosphate, phenyl phosphate and flavin mononucleotide were efficiently hydrolysed by the enzyme and found to be good substrates. Fluoride and tartrate had no inhibitory effect while phosphate, vanadate and molybdate strongly inhibited the enzyme. The acid phosphatase was stimulated in the presence of glycerol, ethylene glycol, methanol, ethanol and acetone, which reflected the phosphotransferase activity. When phosphate acceptors such as ethylene glycol concentrations were increased, the ratio of phosphate transfer to hydrolysis was also increased, demonstrating the presence of a transphosphorylation reaction where an acceptor can compete with water in the rate limiting step involving hydrolysis of a covalent phospho enzyme intermediate. Partition experiments carried out with two substrates, para-nitrophenyl phosphate and phenyl phosphate, revealed a constant product ratio of 1.7 for phosphotransfer to ethylene glycol versus hydrolysis, strongly supporting the existence of common covalent phospho enzyme intermediate. A constant ratio of K (cat)/K (m), 4.3 x 10(4), found at different ethylene glycol concentrations, also supported the idea that the rate limiting step was the hydrolysis of the phospho enzyme intermediate.
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