137 resultados para alkaline proteases
Resumo:
A polyphosphate ester was synthesized by interfacial polycondensation of bisphenol-A and phenylphosphorodichloridate. Accelerated hydrolytic degradation studies were conducted under alkaline conditions. The effect of concentration of alkali and temperature were monitored. The rate of degradation reached a maximum value at 6 molar sodium hydroxide solution and then reduced. The activation energy for hydrolytic degradation was found to be 45 kcal/mol. Diffusion of alkali into the polymer pellet was studied at various concentrations of alkali and at various temperatures. The rate of diffusion also attained a maximum at 6M NaOH and the activation energy for diffusion process was found to be 12 kcal/mol. (C) 2002 John Wiley Sons, Inc.
Isolation and characterization of proteolytic enzymes from the latex of Synadenium grantii Hook, 'f'
Resumo:
Two fractions showing proteolytic enzymes have been obtained from the latex of Synadenium grantii Hook, 'f', using gel-filtration and anion-exchange chromatographic techniques. Both these proteases have the same molecular mass of 76+/-2 kDa each. They exhibit maximal activity at pH 7.0 and at a temperature of 60 degreesC. They display stability over a pH range from 5-10 and are also highly thermostable. Irreversible inhibition by PMSF indicates that they are serine proteases. In addition, histidine residues also appear to play an important role in catalysis as evidenced by inhibition with DEPC. They also exhibit similarity with respect to pH and temperature optima, kinetic properties and thermal stability. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.
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Poly(alkyl aryl ether) dendrimers of up to four generations composed of a phloroglucinol core, branching components, and pentamethylene spacers are synthesized by a divergent growth methodology. A repetitive synthetic sequence of phenolic O-alkylation and O-benzyl deprotection reactions are adopted for the synthesis of these dendrimers. The peripheries of the dendrimers contain 6, 12, 24, and 48 phenolic hydroxyl groups, either in the protected or unprotected form, for the first, second, third, and fourth generations, respectively. Because of the presence of hydrophilic exterior and relatively hydrophobic interior regions, alkaline aqueous solutions of these dendrimers are able to solubilize an otherwise insoluble pyrene molecule and these supramolecular complexes precipitate upon neutralization of the aqueous solutions.
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Trypanosoma evansi is a causative agent of `surra', a common haemoprotozoan disease of livestock in India causing high morbidity and mortality in disease endemic areas. The proteinases released by live and dead trypanosomes entail immunosuppression in the infected host, which immensely contribute in disease pathogenesis. Cysteine proteinases are identified in the infectious cycle of trypanosomes such as cruzain from Trypanosoma cruzi, rhodesain or brucipain from Trypanosoma brucei rhodesiense and congopain from Trypanosoma congelense. These enzymes localised in lysosome-like organelles, flagellar pocket and on cell surface, which play a critical role in the life cycle of protozoan parasites, viz. in host invasion, nutrition and alteration of the host immune response. The paper describes the identification of cysteine proteinases of T. evansi lysate, activity profile at different pH optima and inhibition pattern using a specific inhibitor, besides the polypeptide profile of an antigen. Eight proteinases of T. evansi were identified in the molecular weight (MW) ranges of 28-170 kDa using gelatin substrate-polyacrylamide gel electrophoresis (GS-PAGE), and of these proteinases, six were cysteine proteinases, as they were inhibited by L-3-carboxy-2,3-transepoxypropionyl-lecuylamido (4-guanidino)-butane (E-64), a specific inhibitor. These proteolytic enzymes were most reactive in acidic pH between 3.0 and 5.5 in the presence of dithiothreitol and completely inactive at alkaline pH 10.0. Similarly, the GS-PAGE profile of the serum samples of rats infected with T. evansi revealed strong proteolytic activity only at the 28-kDa zone at pH 5.5, while no proteolytic activity was observed in serum samples of uninfected rats. Further, the other zones of clearance, which were evident in T. evansi antigen zymogram, could not be observed in the serum samples of rats infected with T. evansi. The polypeptide pattern of the whole cell lysate antigen revealed 12-15 polypeptide bands ranging from 28 to 81 kDa along with five predominant polypeptides bands (MW of 81, 66, 62, 55 and 45 kDa), which were immunoreactive with hyperimmune serum (HIS) and serum of experimentally infected rabbits with T. evansi infection. The immunoblot recognised antibodies in experimentally infected rabbits and against HIS as well, corresponding to the zone of clearances at lower MW ranges (28-41 kDa), which may be attributed to the potential of these proteinases in the diagnosis of T. evansi infection. Since these thiol-dependent enzymes are most active in acidic pH and considering their inhibition characteristics, these data suggest that they resemble to the mammalian lysosomal cathepsin B and L.
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The restoration, conservation and management of water resources require a thorough understanding of what constitutes a healthy ecosystem. Monitoring and assessment provides the basic information on the condition of our waterbodies. The present work details the study carried out at two waterbodies, namely, the Chamarajasagar reservoir and the Madiwala Lake. The waterbodies were selected on the basis of their current use and locations. Chamarajasagar reservoir serves the purpose of supplying drinking water to Bangalore city and is located on the outskirts of the city surrounded by agricultural and forest land. On the other hand, Madiwala lake is situated in the heart of Bangalore city receiving an influx of pollutants from domestic and industrial sewage. Comparative assessment of the surface water quality of both were carried out by instituting the various physico–chemical and biological parameters. The physico-chemical analyses included temperature, transparency, pH, electrical conductivity, dissolved oxygen, alkalinity, total hardness, calcium hardness, magnesium hardness, nitrates, phosphates, sodium, potassium and COD measurements of the given waterbody. The analysis was done based on the standard methods prescribed (or recommended) by (APHA) and NEERI. The biological parameter included phytoplankton analysis. The detailed investigations of the parameters, which are well within the tolerance limits in Chamarajasagar reservoir, indicate that it is fairly unpolluted, except for the pH values, which indicate greater alkalinity. This may be attributed to the natural causes and the agricultural runoff from the catchment. On the contrary, the limnology of Madiwala lake is greatly influenced by the inflow of sewage that contributes significantly to the dissolved solids of the lake water, total hardness, alkalinity and a low DO level. Although, the two study areas differ in age, physiography, chemistry and type of inflows, they still maintain a phytoplankton distribution overwhelmingly dominated by Cyanophyceae members,specifically Microcystis aeruginosa. These blue green algae apparently enter the waterbodies from soil, which are known to harbour a rich diversity of blue green flora with several species common to limnoplankton, a feature reported to be unique to the south Indian lakes.Chamarajasagar water samples revealed five classes of phytoplankton, of which Cyanophyceae (92.15 percent) that dominated other algal forms comprised of one single species of Microcystis aeruginosa. The next major class of algae was Chlorophyceae (3.752 percent) followed by Dinophyceae (3.51 percent), Bacillariophyceae (0.47 percent) and a sparsely available and unidentified class (0.12 percent).Madiwala Lake phytoplankton, in addition to Cyanophyceae (26.20 percent), revealed a high density of Chlorophyceae members (73.44 percent) dominated by Scenedesmus sp.,Pediastrum sp., and Euglena sp.,which are considered to be indicators of organic pollution. The domestic and industrial sewage, which finds its way into the lake, is a factor causing organic pollution. As compared to the other classes, Euglenophyceae and Bacillariophyceae members were the lowest in number. Thus, the analysis of various parameters indicates that Chamarajasagar reservoir is relatively unpolluted except for the high percentage of Microcystis aeruginosa, and a slightly alkaline nature of water. Madiwala lake samples revealed eutrophication and high levels of pollution, which is clarified by the physico–chemical analysis, whose values are way above the tolerance limits. Also, the phytoplankton analysis in Madiwala lake reveals the dominance of Chlorophyceae members, which indicate organic pollution (sewage being the causative factor).
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Ever since lysozyme was discovered by Fleming in 1922, this protein has emerged as a model for investigations on protein structure and function. Over the years, several high-resolution structures have yielded a wealth of structural data on this protein. Extensive studies on folding of lysozyme have shown how different regions of this protein dynamically interact with one another. Data is also available from numerous biotechnological studies wherein lysozyme has been employed as a model protein for recovering active recombinant protein from inclusion bodies using small molecules like L-arginine. A variety of conditions have been developed in vitro to induce fibrillation in hen lysozyme. They include (a) acidic pH at elevated temperature, (b) concentrated solutions of ethanol, (c) moderate concentrations of guanidinium hydrochloride at moderate temperature, and (d) alkaline pH at room temperature. This review aims to bring together similarities and differences in aggregation mechanisms, morphology of aggregates, and related issues that arise using the different conditions mentioned above to improve our understanding. The alkaline pH condition (pH 12.2), discovered and studied extensively in our lab, shall receive special attention. More than a decade ago, it was revealed that mutations in human lysozyme can cause accumulation of large quantities of amyloid in liver, kidney, and other regions of gastrointestinal tract. Understanding the mechanism of lysozyme aggregation will probably have therapeutic implications for the treatment of systemic nonneuropathic amyloidosis. Numerous studies have begun to focus attention on inhibition of lysozyme aggregation using antibody or small molecules. The enzymatic activity of lysozyme presents a convenient handle to quantify the native population of lysozyme in a sample where aggregation has been inhibited. The rich information available on lysozyme coupled with the multiple conditions that have been successful in inducing/inhibiting its aggregation in vitro makes lysozyme an ideal model protein to investigate amyloidogenesis.
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The standard Gibbs energies of formation of platinum-rich intermetallic compounds in the systems Pt-Mg, Pt-Ca, and Pt-Ba have been measured in the temperature range of 950 to 1200 K using solid-state galvanic cells based on MgF2, CaF2, and BaF2 as solid electrolytes. The results are summarized by the following equations: ΔG° (MgPt7) = −256,100 + 16.5T (±2000) J/mol ΔG° (MgPt3) = −217,400 + 10.7T (±2000) J/mol ΔG° (CaPt5) = −297,500 + 13.0T (±5000) J/mol ΔG° (Ca2Pt7) = −551,800 + 22.3T (±5000) J/mol ΔG° (CaPt2) = −245,400 + 9.3T (±5000) J/mol ΔG° (BaPt5) = −238,700 + 8.1T (±4000) J/mol ΔG° (BaPt2) = −197,300 + 4.0T (±4000) J/mol where solid platinum and liquid alkaline earth metals are selected as the standard states. The relatively large error estimates reflect the uncertainties in the auxiliary thermodynamic data used in the calculation. Because of the strong interaction between platinum and alkaline earth metals, it is possible to reduce oxides of Group ILA metals by hydrogen at high temperature in the presence of platinum. The alkaline earth metals can be recovered from the resulting intermetallic compounds by distillation, regenerating platinum for recycling. The platinum-slag-gas equilibration technique for the study of the activities of FeO, MnO, or Cr2O3 in slags containing MgO, CaO, or BaO is feasible provided oxygen partial pressure in the gas is maintained above that corresponding to the coexistence of Fe and “FeO.”
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Direct borohydride fuel cells (DBFC) use aqueous alkaline sodium borohydride(NaBH4) as anode fuel to generate electric power with either oxygen or hydrogen peroxide as oxidant. The DBFCs are projected to be very handy for portable power appliances such as laptops and mobile phones in addition to their use in extreme conditions such as underwater and portable military applications. This short review discusses the progress in DBFC research based on electrode materials and membranes.
Resumo:
P-aminobenzoate- intercalated copper hydroxysalt was prepared by coprecipitation at high pH (similar to 12). As the pH was reduced to similar to 7 on washing with water, the development of partial positive charge on the amine end of the intercalated anion caused repulsion between the layers leading to delamination and colloidal dispersion of monolayers of copper hydroxysalt in water. The dispersed copper hydroxysalt monolayers were used as precursors for the synthesis of copper(I)/(II) oxide nanoparticles at room temperature. While the hydroxysalt layers yielded spindle-shaped CuO particles when left to stand, they formed hollow spherical nanoparticles of Cu(2)O when treated with an alkaline solution of ascorbic acid.
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The utility of a soil microbe, namely Bacillus polymyxa, in the removal of organic reagents such as dodecylamine, ether diamine, isopropyl xanthate and sodium oleate from aqueous solutions is demonstrated. Time-bound removal of the above organic reagents from an alkaline solution was investigated under different experimental conditions during bacterial growth and in the presence of metabolites by frequent monitoring of residual concentrations as a function of time, reagent concentration and cell density. The stages and mechanisms in the biodegradation process were monitored through UV-visible and FTIR spectroscopy. Surface chemistry of the bacterial cells as well as the biosorption tendency for various organics were also established through electrokinetic and adsorption density measurements. Both the cationic amines were found to be biosorbed followed by their degradation through bacterial metabolism. The presence of the organic reagents promoted bacterial growth through effective bacterial utilization of nitrogen and carbon from the organics. Under optimal conditions, complete degradation and bioremoval of all the organics could be achieved.
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The technical feasibility of utilization of fly ash as a low-cost adsorbent for the removal of metals from water has been studied. For two types of fly ashes, the retention capacities of copper, lead, and zinc metal ions have been studied. Contact time, initial concentration, and pH have been varied and their effect on retention mechanism has been studied. The dominant mechanisms responsible for retention are found to be precipitation due to the presence of calcium hydroxide, and adsorption due to the presence of silica and alumina oxide surfaces in the fly ash. First-order kinetic plots have revealed that the rate constant increases with increase in the initial concentration and pH. Langmuir adsorption isotherms have been plotted to study the maximum adsorption capacities for metal ions considered under different conditions. X-ray diffraction studies revealed the formation of new peaks corresponding to respective metal ions precipitates under alkaline conditions.
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Background: Development of sensitive sequence search procedures for the detection of distant relationships between proteins at superfamily/fold level is still a big challenge. The intermediate sequence search approach is the most frequently employed manner of identifying remote homologues effectively. In this study, examination of serine proteases of prolyl oligopeptidase, rhomboid and subtilisin protein families were carried out using plant serine proteases as queries from two genomes including A. thaliana and O. sativa and 13 other families of unrelated folds to identify the distant homologues which could not be obtained using PSI-BLAST. Methodology/Principal Findings: We have proposed to start with multiple queries of classical serine protease members to identify remote homologues in families, using a rigorous approach like Cascade PSI-BLAST. We found that classical sequence based approaches, like PSI-BLAST, showed very low sequence coverage in identifying plant serine proteases. The algorithm was applied on enriched sequence database of homologous domains and we obtained overall average coverage of 88% at family, 77% at superfamily or fold level along with specificity of similar to 100% and Mathew's correlation coefficient of 0.91. Similar approach was also implemented on 13 other protein families representing every structural class in SCOP database. Further investigation with statistical tests, like jackknifing, helped us to better understand the influence of neighbouring protein families. Conclusions/Significance: Our study suggests that employment of multiple queries of a family for the Cascade PSI-BLAST searches is useful for predicting distant relationships effectively even at superfamily level. We have proposed a generalized strategy to cover all the distant members of a particular family using multiple query sequences. Our findings reveal that prior selection of sequences as query and the presence of neighbouring families can be important for covering the search space effectively in minimal computational time. This study also provides an understanding of the `bridging' role of related families.
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Titanium carbide (TiC) possesses fascinating properties like high electrical conductivity and high mechanical strength coupled with high corrosion resistance and stability in acidic and alkaline environments. The present study demonstrates the tunability of mechanistic aspects of oxygen reduction reaction (ORR) using TiC nanostructures. One dimensional TiC nanostructures (TiC-NW) have been synthesized using a simple, hydrothermal method and used as a catalyst for ORR. Shape dependent electroactivity is demonstrated by comparing the activity of TiC-NW with its bulk counterparts. Comparative studies reveal higher ORR activities in the case of 1D TiC-NW involving similar to 4 electrons showing efficient reduction of molecular oxygen. Excellent stability and high methanol tolerance with good selectivity for ORR is reported.
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Phosphogypsum is added to building materials to accelerate fly ash pozzolanic reaction and contributes to early strength development of concrete. The release of unacceptable fluoride levels by phoshogypsum on contact with water is a major impediment in its usage to manufacture building products because excess fluoride consumption causes dental and skeletal fluorosis. This paper examines the efficacy of fly ash pozzolanic reactions in controlling fluoride release by phosphogypsum. Fly ash (FA), sand (S), lime (L), and phosphogypsum (G) (FA-S-L-G) slurries are cured for various periods, and the fluoride released by the mix is monitored as a function of time. A substantial reduction in fluoride release was observed and is attributed to entrapment of phosphogypsum particles in a cementious matrix formed by fly ash-lime pozzolanic reactions coupled with consumption of fluoride in formation of insoluble compounds. The compressive strength developed by compacted FA-S-L-G specimens with time was observed to be a three-stage process; maximum strength mobilization occurred during 14 and 28days of curing at room temperature. Exposure of the compacted FA-S-L-G specimens to acidic and alkaline environments for 9 days did not impact their compressive strengths. (C) 2013 American Society of Civil Engineers.
Resumo:
Electrodeposition of Au on poly (3,4-ethylenedioxythiophene) (PEDOT) coated carbon paper electrode results in the formation of a stable 3-D urchin-like morphology. Au-PEDOT/C electrode exhibits higher surface area, greater catalytic activity, higher sensitivity and lower detection limit for glucose analysis in an alkaline medium than Au/C electrode. Au-PEDOT/C electrode exhibits a linear current response in glucose concentration ranging up to 10 mu M with sensitivity of 515 mu A cm(-2) mu M-1 (on the basis of geometric area) and a low detection limit of 0.03 mu M with signal to noise ratio of 3. Thus, the PEDOT under-layer improves the property of Au for glucose analysis. (c) 2013 The Electrochemical Society.